U.S. patent application number 14/090317 was filed with the patent office on 2014-07-03 for thin-film layered centrifuge device and analysis method using the same.
This patent application is currently assigned to SAMSUNG ELECTRONICS CO., LTD.. The applicant listed for this patent is SAMSUNG ELECTRONICS CO., LTD.. Invention is credited to Jae Chern YOO.
Application Number | 20140186935 14/090317 |
Document ID | / |
Family ID | 40901539 |
Filed Date | 2014-07-03 |
United States Patent
Application |
20140186935 |
Kind Code |
A1 |
YOO; Jae Chern |
July 3, 2014 |
THIN-FILM LAYERED CENTRIFUGE DEVICE AND ANALYSIS METHOD USING THE
SAME
Abstract
Disclosed herein is a thin-film layered centrifuge device and an
analysis method using the same. One example of an embodiment of the
present invention is a thin film layered centrifuge device where a
device, such as a lab on a chip, a protein chip and a DNA chip, for
diagnosing and detecting a small amount of material in a fluid is
integrated into a rotatable thin-film layered body, and to an
analysis method using the thin-film layered centrifuge device.
Inventors: |
YOO; Jae Chern; (Pohang-si,
KR) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
SAMSUNG ELECTRONICS CO., LTD. |
Suwon-si |
|
KR |
|
|
Assignee: |
SAMSUNG ELECTRONICS CO.,
LTD.
Suwon-si
KR
|
Family ID: |
40901539 |
Appl. No.: |
14/090317 |
Filed: |
November 26, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
12863684 |
Jul 20, 2010 |
|
|
|
PCT/KR09/00306 |
Jan 21, 2009 |
|
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14090317 |
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Current U.S.
Class: |
435/287.2 ;
422/69; 435/287.7 |
Current CPC
Class: |
G01N 2035/00247
20130101; G01N 2035/00495 20130101; G01N 33/5302 20130101; G01N
35/00069 20130101; G01N 35/00029 20130101; B01L 3/502753
20130101 |
Class at
Publication: |
435/287.2 ;
422/69; 435/287.7 |
International
Class: |
G01N 35/00 20060101
G01N035/00; B01L 3/00 20060101 B01L003/00; G01N 33/53 20060101
G01N033/53 |
Foreign Application Data
Date |
Code |
Application Number |
Jan 21, 2008 |
KR |
10-2008-0006890 |
Claims
1. A centrifuge device comprising: A rotatable body; a sample inlet
to inject a sample; a sample chamber to store the sample injected
into the sample inlet; a specimen chamber to store a specimen
obtained from the sample during centrifugation; one or more assay
sites in which a capture probe to be bound to the specimen is
immobilized and/or reagent for biochemical reactions with the
specimen is stored; a liquid passage, to provide a passage to
connect the specimen chamber to the assay sites, allowing the
specimen retained in the specimen chamber during centrifugation to
flow to the assay site by the fluid flow device upon non-rotation
of the body; and an absorbent pad is provided between the terminal
of the liquid passage and an inlet of the assay site.
2. The device according to claim 1, wherein the liquid passage
comprises an inward channel and an outward channel to form a U- or
V-shape.
3. The device according to claim 1, further comprising a trash
chamber is connected the one side of the assay site to collect
debris not bound to the capture probe by a washing process.
4. The device according to claim 1, further comprising a remnant
chamber to store a remnant rather than specimens produced during
centrifugation.
5. The device according to claim 1, further comprising a set-amount
channel and an excess chamber to store excess specimen or sample
present in the specimen chamber.
6. The device according to claim 4, wherein the remnant chamber is
a capillary tube chamber.
7. The device according to claim 1, further comprising one or more
chambers selected from the group consisting of cleaning chambers,
mixing chambers, buffer chambers and substrate chambers.
8. The device according to claim 4, further comprising a bottle
neck channel to connect the specimen chamber to the remnant
chamber, wherein the bottle neck channel comprises two or more thin
film channels.
9. The device according to claim 8, wherein the thin film channels
are formed between base material layers to constitute the body by a
channel-shaped thin film adhesive tape.
10. The device according to claim 1, wherein the body comprises an
upper base material, an intermediate base material and a lower base
material which are laminated in this order and adhered to one
another, wherein the body further comprises: a first thin film
adhesive tape laminated between the upper base material and the
intermediate base material, to adhere the upper base material to
the intermediate base material; and a second thin film adhesive
tape laminated between the intermediate base material and the lower
base material, to adhere the intermediate base material to the
lower base material.
11. The device according to claim 10, wherein the substrate is
composed of at least one selected from the group consisting of
hydrophobic materials, silicon wafers, polypropylene, polyacylate,
polyvinylalcohol, polyethylene, polymethyl methacrylate (PMMA),
cyclic olefin copolymers (COCs) and polycarbonate.
12. The device according to claim 1, wherein the assay site
comprises a porous membrane or a strip in which the capture probe
is fixed.
13. The device according to claim 1, wherein the body comprises a
wireless RF IC having one or more functions selected from the group
consisting of temperature measurement, assay site detection,
storage and transmission of assay site detection results, personal
privacy encryption, identification (ID) storage and transmission of
the thin film centrifuge device, test date storage and efficient
period storage.
14. The device according to claim 7, further comprising: magnetic
micro-beads contained in the mixing chamber; a slider movable in a
lower part of the body; and a permanent magnet mounted on the
slider, to apply attraction force to the magnetic micro-beads and
thus move the magnetic micro-beads, wherein the magnetic
micro-beads are moved in accordance with movement of the slider to
induce mixing of liquids in the mixing chamber.
15. The device according to claim 7, further comprising: magnetic
micro-beads contained in the mixing chamber; a slider movable in a
lower part of the body; and a permanent magnet mounted on the
slider, to attract the magnetic micro-beads and thus move the
magnetic micro-beads, wherein the permanent magnet is maintained on
the corresponding diameter in the mixing chamber and the body is
rotated to induce movement of the magnetic micro-beads, thereby
mixing of the liquids in the mixing chamber.
16. The device according to claim 1, further comprising a
set-amount chamber coated with a super-hydrophilic material and a
concentric channel coated with a super-hydrophilic material,
wherein the set-amount chamber is interposed between the concentric
channel and the assay site, wherein the concentric channel is
connected to an outlet of the liquid valve, the device further
comprising an overflow chamber to allow the specimen to remain in
the set-amount chamber by rotation of the body and the specimen in
the concentric channel to be extracted by centrifugal force and
thereby store the residual specimen, after the set-amount chamber
and the concentric channel are filled with the specimen, while the
specimen in the specimen chamber hydrophilic-flows through the
concentric channel.
17. The device according to claim 16, wherein the specimen of the
set-amount chamber overcomes the fluid flow barrier formed between
the set-amount chamber and the assay site by centrifugal force
generated by rotation of the body and then flows in the assay
site.
18. The device according to claim 8, wherein the remnant chamber
comprises neither a channel nor an outlet to allow liquids to flow
in or leak out, except the bottle neck channel.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a Continuation application of U.S.
application Ser. No. 12/863,684 filed on Jul. 20, 2010 and claims
the benefit of Korean Patent Application No. 10-2008-0006890 filed
Jan. 21, 2008 in the Korean Intellectual Property Office and
PCT/KR/2009/00306 filed Jan. 21, 2009 in the Korean Intellectual
Property Office, the disclosure of which is incorporated herein by
reference.
BACKGROUND
[0002] 1. Field
[0003] Embodiments of the present invention relate to a thin film
layered centrifuge device and an analysis method using the same.
More specifically, embodiments of the present invention relate to a
thin film layered centrifuge device wherein a thin-film rotatable
body is integrated with an apparatus to diagnose and detect a small
amount of materials present in fluids, such as a lab-on-a-chip, a
protein chip and a DNA chip, and an analysis method using the
same.
[0004] 2. Description of the Related Art
[0005] General clinical diagnosis and analysis apparatuses to
detect a small amount of analytes in fluids include multiple sample
arrangement and automated sample feed devices, and have a structure
in which an apparatus to analyze a large number of test samples in
series or parallel is integrated on a rotatable thin film body to
improve analysis efficiency and economic efficiency. For example,
rotatable bio discs are one type of such analyzers. Such thin-film
analyzers for clinical tests enable various types of analysis to be
performed accurately and automatically at a low cost, based on the
centrifugal force generated by rotation of the bio disc, with a
small amount of samples and specimens.
[0006] Considering thin film-type CDs and DVDs, standard compact
discs can be formed from a 12 cm polycarbonate substrate,
reflective metal layer and a protective layer coating. The format
of CDs, DVD and CD-ROMs may be in accordance with ISO 9660
industrial standard. The polycarbonate substrate is made of
optical-quality transparent polycarbonate. A data layer in standard
printed or bulk-copied CDs is a part of the polycarbonate substrate
and the data is printed by a stamper in the form of a series of
pits during injection molding. Polycarbonate molten during the
injection molding process is injected into a mold at a high
pressure and is then cooled to obtain polycarbonate in the form of
the mold, the stamper or a mirror-image thereof, and pits showing
binary data on the disc substrate are formed on the polycarbonate
substrate. The stamping master may be a glass. Such a disc may be
modified into a thin film analyzer to diagnose and detect a small
amount of material in a fluid. In this case, channels to allow flow
of fluids, chambers to store buffer solutions, and holes or valves,
may be formed on the surface of the disc instead of the pits.
[0007] Hereinafter, a disc wherein bio chips such as a
lab-on-a-chip, a protein chip and DNA chip to diagnose and detect a
small amount of materials in fluids are integrated in a disc such
as a conventional CD-ROM or DVD, or a disc to perform biological
and chemical processes to diagnose and detect a small amount of
materials in fluids is referred to as a bio disc.
[0008] Conventional bio discs may include a plurality of chambers
to store a large volume of liquid-phase biological and chemical
materials required for chemical processes. The biological and
chemical processes include preparing specimens from samples,
centrifugation, DNA amplification, hybridization, antigen-antibody
reactions, mixing, washing and the like. The biological and
chemical processes may be sequentially automatically performed on
the bio disc, which is known in the art. However, there is a need
to solve the following problems associated with bio discs in order
to make practical application possible.
[0009] For the process of centrifugation wherein specimens are
extracted from samples, a valve, which does not leak during
centrifugation, is required. Conventional valves to realize
close/open operations based on physical movement perform the
operations in such a manner that a ball or closing portion comes
into contact with a hole or a channel, or the ball or closing
portion is separated therefrom, which are known in the art.
However, these valves inevitably allow opening based on physical
movement, thus entailing incomplete closing. Accordingly, inherent
hydraulic pressure of fluids may cause leakage during
centrifugation. This leakage prevents extraction of desired amounts
of specimens from samples by centrifugation, thus causing
deterioration in assay reliability and accuracy. Accordingly, there
is a need for a centrifuge device that does not leak during rapid
rotation.
SUMMARY
[0010] Therefore, it is one aspect of the present invention to
provide a thin film layered centrifuge device wherein a thin-film
rotatable body is integrated with an apparatus to diagnose and
detect a small amount of materials present in fluids, such as a
lab-on-a-chip, a protein chip and a DNA chip, and an analysis
method using the same. Embodiments provide a thin film centrifuge
device in which bio chips, such as a lab-on-a-chip, protein chips
and DNA chips, to diagnose and detect a material in a fluid are
integrated, by providing a thin film body with a centrifuge device
causing no leakage, when specimens are extracted from samples by
centrifugation, and an analysis method using the same.
[0011] Additional aspects of the invention will be set forth in
part in the description which follows and, in part, will be obvious
from the description, or may be learned by practice of the
invention.
[0012] In accordance with one aspect of the present invention,
provided is a thin film centrifuge device including: a sample inlet
to inject a sample; a sample chamber to store the sample injected
into the sample inlet; a specimen chamber to store a specimen
obtained from the sample during centrifugation; a remnant chamber
to store a remnant rather than specimens produced during
centrifugation; a bottle neck channel to connect the specimen
chamber to the remnant chamber; one or more assay sites in which a
capture probe to be bound to the specimen is immobilized and/or
reagent for biochemical reactions with the specimen is stored; a
trash chamber to collect debris not bound to the capture probe by a
cleaning process; a rotatable hydrophobic body in which the sample
inlet, the sample chamber, the specimen chamber, the remnant
chamber, the trash chamber, the bottle neck channel and the assay
site are integrated; one or more fluid flow devices to transfer the
specimen from the specimen chamber to the assay site upon
non-rotation of the body, the fluid flow devices selected from the
group consisting of a hydrophilic fluid flow pump, a chamber pump,
an erythrocyte pump and an absorption pump; and a liquid valve
coated with a super-hydrophilic material, to provide a passage to
connect the specimen chamber to the assay sites, allowing the
specimen retained in the specimen chamber during centrifugation to
flow to the assay site by the fluid flow device upon non-rotation
of the body.
[0013] The liquid valve has a U- or V-shape to provide a
hydrophilic channel to connect the specimen chamber to the assay
site, when the body creases rotation, and at the same time to
prevent transfer of the fluid in the specimen chamber to the assay
site when the body rotates. The surface of the liquid valve is
treated with a super-hydrophilic material and the fluid trapped in
the specimen chamber when the body rotates can be transferred to
the assay site via hydrophilic fluid flow by the liquid valve when
the body ceases rotation. In one embodiment, by using the channel
having a superhydrophilic-coated U- or V-shape channel as the
liquid valve, all the specimens in the specimen chamber can
hydrophilic-flow to the assay site, when the body ceases
rotation.
[0014] In one embodiment, volume of the total specimen in the
specimen chamber, required for quantitative analysis can be
determined. Accordingly, the embodiment comprises transfer of the
total specimen from the specimen chamber to the assay site.
However, the specimen present in the specimen chamber, having a
viscosity comparable to blood serum may be partially transferred
through the U- or V-shaped channel to the assay site due to
inherent viscosity. That is, when the body ceases rotation, through
the U- or V-shaped hydrophilic channel, only a part of the specimen
in the specimen chamber may be transferred to the assay site, thus
making quantitative analysis impossible. Accordingly, in one
embodiment, in order to transfer the total specimen from the
specimen chamber to the assay site, when the body ceases rotation,
a fluid flow means may be provided to the specimen in the specimen
chamber.
[0015] In one embodiment, the specimen in the specimen chamber can
be entirely transferred to the assay site using the following four
fluid flow means.
[0016] First, the remnant chamber swollen while the body rotates is
returned, and, at the same time, generates air pressure, when the
body ceases rotation. This air pressure may generate fluid-driving
force to transfer the total specimen from the specimen chamber to
the assay site. Hereinafter, a fluid flow means based on
fluid-driving force generated by swelling and return of the remnant
chamber is referred to as a chamber pump. When the body rotates,
the swelling of the remnant chamber may be caused by centrifugal
force. The remnant chamber is arranged farther from the
circumference than the specimen chamber and stores a material
heavier than the specimen, for example, erythrocyte, according to
one embodiment, and the upper base material of the remnant chamber
may swell during this high-speed rotation. The upper base material
of the remnant chamber may be a thin film having a thickness of 0.1
mm to 0.6 mm, enabling easy swelling during high-speed
rotation.
[0017] Second, the material in the remnant chamber compressed while
the body rotates, for example, erythrocyte according to one
embodiment, is expended and at the same time, generates a fluid
pressure, when the body creases rotation. The fluid pressure
specimen chamber may generate fluid-driving force to transfer the
total specimen from the specimen chamber to the assay site.
Hereinafter, fluid flow means using fluid-driving force based on
compression and expansion of erythrocyte in the remnant chamber is
referred to as an erythrocyte pump. When the body rotates,
compression of erythrocyte in the remnant chamber may be generated
by centrifugal force.
[0018] Third, an absorption force to absorb the specimen having
reached the terminal of the liquid valve through the U- or V-shaped
hydrophilic channel including an absorbent pad, a sample pad, or a
super-hydrophilic chamber between the terminal of the liquid valve
and an inlet of the assay site continuously generates fluid-driving
force to transfer the total specimen from the specimen chamber to
the assay site or super-hydrophilic chamber. Hereinafter, a fluid
flow means using fluid-driving force based on absorption force of
the absorbent pad or the sample pad or hydrophilic absorption force
of the super-hydrophilic chamber is referred to as an absorption
pump.
[0019] Fourth, by coating the U- or V-shaped channel with a
super-hydrophilic material, hydrophilic absorption force to
transfer the specimen from the specimen chamber to the assay site
can generate fluid-driving force to transfer the total specimen
from the specimen chamber to the assay site. Hereinafter, a fluid
flow means based on fluid-driving force derived from the
hydrophilic absorption force is referred to as hydrophilic fluid
flow.
[0020] In one embodiment, by transferring the total specimen in the
specimen chamber to the assay site via the fluid flow means, the
specimen chamber can be emptied. After the total specimen is
discharged from the specimen chamber through the liquid valve to
the assay site, the fluids are not transferred from the remnant
chamber to the liquid valve due to strong capillary action of the
bottle neck channel to fluids. That is, strong capillary action of
the bottle neck channel to fluids may be equivalent to fluid flow
force of the fluid flow means, preventing further transfer of the
fluids to the assay site. Accordingly, only quantitative specimen
is transferred to the assay site.
[0021] In one embodiment, the chamber pump, the erythrocyte pump,
the absorption pump and fluid flow means via hydrophilic fluid flow
may further utilize fluid-driving force based on capillary force
obtained from the U- or V-shaped channel.
[0022] In one embodiment, the sample chamber may be coated with a
super-hydrophilic material. In one embodiment, the
super-hydrophilic coating includes hydrophilic coating.
[0023] In one embodiment, the sample includes various biomaterials,
for example, blood. In addition, the specimen includes substances
obtained from a sample by centrifugation, for example, blood serum
or plasma obtained from blood.
[0024] In exemplary embodiments, the term "blood serum" used herein
is intended to include the blood serum, plasma and leukocytes.
[0025] In one embodiment, the remnant chamber may be a capillary
tube chamber.
[0026] When blood is centrifuged, it is separated into blood serum,
blood clotting factors, plasma and erythrocytes. The blood clotting
factors may be mostly erythrocyte. Accordingly, when blood of the
sample chamber is stored, and the specimen chamber and the remnant
chamber are centrifuged, blood serum is left in the specimen
chamber and erythrocyte is left in the remnant chamber. In this
case, when the rotation is ceased after centrifugation, erythrocyte
may be admixed with blood serum again. That is, rotation of the
body should be stopped in order to extract only blood serum after
centrifugation. In this case, erythrocyte is admixed with blood
serum again, making extraction of only blood serum difficult.
Accordingly, in one embodiment, the remnant chamber is provided as
a capillary tube chamber having a low height (narrow), to allow
erythrocyte to remain in the remnant chamber based on capillary
action or bonding force between the surface of the remnant chamber,
thereby preventing re-admixing of erythrocyte with blood serum. The
bonding force between the surface of the remnant chamber and
erythrocyte is based on strong viscosity of the erythrocyte. As a
result, centrifuged erythrocyte is not admixed with blood serum and
is thus left in the remnant chamber although the body ceases
rotation. The height of the capillary tube chamber may be, for
example, 0.1 mm to 0.6 mm.
[0027] In one embodiment, the body may further comprise a cleaning
chamber to store a cleaning solution required for cleaning.
[0028] In one embodiment, the body may further comprise a mixing
chamber to mix the two fluids.
[0029] In one embodiment, the body may further comprise a buffer
chamber to store a dilution buffer to dilute the specimen or a
label to be linked to a target material in the specimen. The label
may have antibody- or DNA-linked chromatic particles such as gold
or gold conjugates, latex or fluorescent labels, radioisotopes,
enzymes, or enzyme-linked antibody labels. The enzyme may render
color using a substrate solution reacted with an enzyme.
[0030] In one embodiment, the body may further comprise a substrate
chamber to store the substrate solution reacted with the
enzyme.
[0031] In one embodiment, the specimen comprises biomaterials
participating in biochemical bonding, such as blood serum, DNA,
proteins, ligands or receptors.
[0032] In one embodiment, the thin film centrifuge device may
further comprise a thin film cylindrical magnet to perform
azimuthal direction search of the assay site in the body. Instead
of the thin film cylindrical magnet, thin-film ferromagnetic metal
particles may be used. The thin film cylindrical magnet or thin
film ferromagnetic metal particles may have a diameter of 1 mm to 5
mm and a thickness of 0.1 mm to 1 mm.
[0033] In one embodiment, the bottle neck channel may be composed
of two thin film channels. The bottle neck channel provides a
passage to transfer the remnant from the specimen chamber to the
remnant chamber, or to transfer the centrifuged analyte from the
remnant chamber to the specimen chamber, while the sample in the
specimen chamber and the sample in the remnant chamber are
independently centrifuged by centrifugal force generated by
rotation of the body. That is, the bottle neck channel may provide
a passage, allowing the analyte and remnant separated during
centrifugation to be transferred from the specimen chamber to the
remnant chamber.
[0034] In one embodiment, the remnant chamber has no outlet. That
is, the remnant chamber has no channel or outlet, to allow liquids
to flow in or leak out, except the bottle neck channel. The bottle
neck channel is provided in a thin film channel, preventing return
of the remnant from the remnant chamber to the specimen chamber
when the body ceases rotation, and maintaining a predetermined
amount of the specimen in the specimen chamber. That is, when the
body stops, strong capillary action of the bottle neck channel
composed of the thin film channel and absence of outlet in the
remnant chamber make it impossible to freely transfer the fluids
from the remnant chamber to the specimen chamber. In addition,
after the total specimen is discharged from the specimen chamber
through the liquid valve via hydrophilic fluid flow, the fluids in
the remnant chamber do not transfer to the liquid valve due to
strong capillary action of the bottle neck channel to fluids. That
is, strong capillary action of the bottle neck channel to fluids is
equivalent to the force of hydrophilic fluid flow to induce
transfer to the liquid valve, preventing further transfer of fluids
to the liquid valve.
[0035] In one embodiment, the hydrophilic channel may be treated
via surface modification using a porous material, or coated with a
water-based paint or a super-hydrophilic paint.
[0036] In one embodiment, the thin film centrifuge device may
further comprise a spindle motor to rotate the body.
[0037] The thin film centrifuge device according to one embodiment
comprises a bio pickup optical module (BOPM) mounted on a slider
and a slide motor to control movement of the BOPM, enabling space
addressing of the chambers. A laser beam generator and a permanent
magnet are mounted on the BOPM, and coordinates of the BOPM may be
moved or controlled by control of the slide motor. The laser beam
generator uses, for example, an optical pick-up device. The radial
direction search may be carried out by control of the slide motor.
The azimuthal direction search is carried out by rotating the body
to some extent, while controlling short rotation of the spindle
motor or the stepper motor, when the slider is stopped. The stepper
motor may be connected or mounted to a gear on the shaft of the
spindle motor for azimuthal direction rotation of the body.
[0038] The thin film centrifuge device may further comprise a
temperature-control means to control reaction temperatures of the
chambers. The temperature-control means may comprise at least one
selected from the group consisting of temperature-measurement
means, heating means and cooling means. The heating means comprises
a laser beam generator mounted on the slider. The cooling means may
perform rotation-cooling using rotation of body. Heat emission
efficiently occurs due to contact between the surface of the
chamber and air during rotation of the body. The temperature
measurement means may measure the temperature of the corresponding
chamber using the temperature sensor connected to the wireless RF
IC provided in the body and wirelessly transmit the temperature to
the outside central control device.
[0039] The body comprises a rotatable thin film disc, composed of
an upper base material, an intermediate base material and a lower
base material. For example, the disc has a diameter of 120 mm, 80
mm or 32 mm, a thickness of 0.6 mm to 3 mm and a circular
shape.
[0040] The flow of fluids may be carried out by centrifugal force
or capillary action generated by rotation force of the body
phenomenon, or using a super-hydrophilic-coated channel.
[0041] The body may be selected from various materials such as
plastics, glasses, silicon wafers or hydrophobic materials.
However, plastics are preferred, owing to economical efficiency,
processability, and compatibility with conventional laser
reflection-based detectors such as CD-ROM and DVD detectors. The
substrate is composed of at least one selected from the group
consisting of silicon wafers, polypropylene, polyacylate,
polyvinylalcohol, polyethylene, polymethyl methacrylate (PMMA),
cyclic olefin copolymers (COCs) and polycarbonate. In addition, the
body may be coated with aluminum to prevent evaporation of liquids
stored in the chamber.
[0042] The body may be composed of an upper base material, an
intermediate base material and a lower base material. These
materials may be adhered to one another by an adhesive agent. The
adhesive agent may be prepared from a material selected from the
group consisting of silicon, rubbers, modified silicon, acrylic,
polyester and epoxy.
[0043] The body is composed of an upper base material, an
intermediate base material and a lower base material which are
laminated in this order and adhered to one another and further
comprises a first thin film adhesive to adhere the upper base
material to the intermediate base material, and a second thin film
adhesive to adhere the intermediate base material to the lower base
material. The thin film adhesive tape may be a one-side tape or
two-side tape. The tape is obtained by surface-treating one or both
sides of release papers such as papers, vinyl polyester films,
polyethylene films and other synthetic materials with an adhesive
(or a gluing) agent. According to requirements, adhesive materials
exhibiting properties such as superior sealing, buffering,
vibration reduction, impact resistance, heat resistance, absorbent
performance or adhesion force may be used. The thin film adhesive
tape may be prepared via thin film coating on one side of a
substrate using an adhesive agent, performed by adhering an
one-side tape to the substrate and removing a release paper
therefrom, or printing a dispenser, spraying or silk screen
printing on one side of the substrate. That is, in one embodiment,
the thin film adhesive tape may be coated on the substrate in the
form of a thin film using an adhesive (gluing) agent without any
release paper.
[0044] The device may further comprise magnetic micro-beads
contained in the mixing chamber; a slider movable in a lower part
of the body; and a permanent magnet mounted on the slider, to apply
attraction force to the magnetic micro-beads and thus move the
magnetic micro-beads, wherein the magnetic micro-beads are moved in
accordance with movement of the slider to induce mixing of liquids
in the mixing chamber.
[0045] In another embodiment, the device may further comprise
magnetic micro-beads contained in the mixing chamber; a slider
movable in a lower part of the body, and a permanent magnet mounted
on the slider, to apply attraction force to the magnetic
micro-beads and thus move the magnetic micro-beads, wherein the
magnetic micro-beads are moved in accordance with movement of the
slider, by keeping the permanent magnet on the corresponding
diameter of the mixing chamber and rotating the body, to induce
mixing of liquids in the mixing chamber.
[0046] In one embodiment, the mixing operation may be performed
prior to radial direction search, or radial direction search and
azimuthal direction search of the mixing chamber to perform the
mixing operation.
[0047] In one embodiment, the specimen chamber may further comprise
a set-amount channel connected to the remnant chamber, to transfer
excess fluid.
[0048] The assay site may store reagents for biochemical reaction
or include a porous membrane on which a capture probe is fixed. The
assay site may comprise a porous membrane; a capture probe fixed on
the porous membrane; a capture probe spotted and fixed in an array
on the substrate; micro-pores formed on the substrate, or a capture
probe fixed on the micro-pores. The assay site comprises a porous
membrane and line- or spot-shaped tumor markers or disease labels
fixed on the porous membrane as a test line, the porous membrane
may be in the form of a strip, allowing lateral flow or flow
through. The porous membrane is provided at one terminal with a
sample pad and a conjugate pad, at the other terminal with an
absorbent pad. The tumor label or disease label may be AFP, PSA,
CEA, CA19-9, CA125, CA15-3 or Alzheimer markers, or myocardial
infarction label.
[0049] The assay site may further comprise a capture probe for a
reference line and a control line fixed on the porous membrane. The
reaction concentration of the reference line may be a cutoff value.
For example, the cutoff concentration of the reference line may be
3 ng/ml, 4 ng/ml, 10 ng/ml, 20 ng/ml, 30 ng/ml, 40 ng/ml or 50
ng/ml. For example, qualitative or quantitative analysis can be
realized based on the difference in reaction intensity between the
reference line and the test line. For example, qualitative or
quantitative analysis can be realized based on the difference in
reaction intensity between the background of the strip and the test
line. For example, qualitative or quantitative analysis can be
realized by determining reaction intensity of test lines using the
linear function of reaction intensity formed by the plurality of
reference lines. Qualitative or quantitative analysis can be
realized by determining reaction intensity of test lines using a
linear function of reaction intensity formed by the reference line
and the control line.
[0050] The body may comprise a wireless RF IC to measure
temperature, detect assay sites, store and transmit the assay site
detection results and perform personal privacy encryption. The thin
film centrifuge device may further comprise a detector to detect
the reaction results of the assay site. The detector may be a
spectrometer including a light source and a photodetector. In
addition, the detector may be an optical measurement apparatus
including an illuminator and an image sensor (for example, CCD,
CMOS, or CIS sensors). Alternatively, the detector may be a
photometric measurement apparatus comprising a laser beam apparatus
and a photodetector.
[0051] The thin film layered centrifuge device and an analysis
method using the same according to one embodiment may be applied to
thin-film devices to diagnose and detect a small amount of
biological and/or chemical materials in fluids such as a
lab-on-a-chip, protein chips and DNA chips. For example, the thin
film centrifuge device may be integrated in thin film discs such as
conventional CD-ROMs or DVDs.
[0052] The thin film layered centrifuge device and an analysis
method using the same according to one embodiment may be applied to
lab-on-a-chips utilizing ELISA/CLISA analysis methods,
lab-on-a-chips utilizing rapid test methods; or thin-film devices
to diagnose and detect a small amount of biological and/or chemical
materials in fluids, such as lab-on-a-chips for food
poisoning-causing bacteria assays, residual antibiotic assays,
residual pesticide assays, genetically modified food tests, food
allergy tests, contaminant assays or paternity tests, and meat
types and origin region tests. The residual pesticide comprises
pesticide contained in vegetables or fruits, for example, the
most-generally used organophosphorus and carbamate
insecticides.
[0053] In one embodiment, the biomaterial may be at least one
selected from DNA, oligonucleotides, RNA, PNA, ligands, receptors,
antibodies, antibodies, milk, urine, saliva, hairs, crop samples,
meat samples, fish samples, bird samples, wastewater, livestock
samples, food samples, oral cells, tissue samples, semen, proteins
or other biomaterials.
[0054] Upon urine assay, the thin film centrifuge device may
perform leukocyte, blood, protein, nitrite, pH, specific gravity,
glucose, ketone, ascorbic acid, urobilinogen and bilirubin
assays.
[0055] Hair assays accurately measure historical record of
nutriment and toxic substances including minerals accumulated in
blood or urine assays. Hair assays accurately detect oversupply and
lack of inorganic materials for a long time and provide a standard
to assay the amount of toxic heavy metals, which is known in the
art.
[0056] In accordance with another aspect of the present invention,
provided is an analysis method using a thin film centrifuge device
including: a sample inlet to inject a sample; a sample chamber to
store the sample injected into the sample inlet; a specimen chamber
to store a specimen obtained from the sample during centrifugation;
a remnant chamber to store a remnant rather than specimens produced
during centrifugation; a bottle neck channel to connect the
specimen chamber to the remnant chamber; an excess chamber to store
excess specimen exceeding a predetermined amount of the specimen
chamber; one or more assay sites in which a capture probe to be
bound to the specimen is immobilized and/or reagent for biochemical
reactions with the specimen is stored; a liquid valve coated with a
super-hydrophilic material, the liquid valve formed on a passage to
connect the specimen chamber to the assay sites; a trash chamber to
collect debris not bound to the capture probe by a cleaning
process; and a rotatable hydrophobic body in which the sample
inlet, the sample chamber, the specimen chamber, the excess
chamber, the trash chamber, the bottle neck channel and the assay
sites are integrated; the method including: injecting a sample into
the sample chamber through the sample inlet; transferring the
sample from the sample chamber to the specimen chamber and the
remnant chamber based on centrifugal force generated by rotation of
the body and moving the residual sample to the remnant chamber,
when the sample is present in an amount exceeding a predetermined
level of the specimen chamber; centrifuging the sample in the
specimen chamber and the remnant chamber based on centrifugal force
generated by rotation of the body, and at the same time,
transferring the remnant present in the specimen chamber through
the bottle neck channel to the remnant chamber, or the specimen
from the remnant chamber through the bottle neck channel to the
specimen chamber; hydrophilic-flowing the specimen retained in the
specimen chamber through the liquid valve to the assay site when
the body ceases rotation; and binding the specimen present in the
assay site to the capture probe present in the assay site, or
biochemically reacting the specimen with a reagent in the assay
site.
[0057] The analysis method may further include adding the cleaning
solution to clean the assay site, and drying and dehydrating the
assay site.
[0058] The analysis method may further include one or more
operations selected from: searching an assay site; qualitatively or
quantitatively analyzing reaction results of the assay site;
allowing the wireless RF IC integrated in the body to detect the
assay site to realize wireless transmission; displaying diagnosis
results derived from the analysis on a computer monitor; and
remote-transmitting diagnosis results and questionnaires to a
doctor connected through an internet network; or obtaining the
doctor's prescription.
[0059] The analysis method may further include moving magnetic
micro-beads in the mixing chamber via magnetic force to mix the
liquid in the mixing chamber.
[0060] The analysis method may further include preventing fluid
leakage from the specimen chamber by centrifugal force using a
liquid valve having a U- or V-shape based on the rotation center of
the body, while the body rotates.
[0061] The analysis method may further include controlling the
temperature of the assay site using the temperature-control
means.
[0062] The analysis method may further include searching a specific
one selected from the plurality of assay sites and selecting the
same; and detecting the response of the specific assay site. The
detection process may be carried out using a spectrometer. The
detection of the assay site using the spectrometer may be carried
out after searching the chamber through controlling the rotation
angle of the body using the stepper motors or gears connected
thereto or through the azimuthal valve search process, or by
sequentially measuring light absorption in the respective assay
sites during rotation of the body through space-addressing assay
sites using a blank solution chamber during rotation of the
body.
[0063] The light source or light source device of the spectrometer
may be a white light LED, an RGB laser, or an LD module in which a
plurality of laser diodes (LDs) are integrated.
[0064] The detection of the assay site using the spectrometer may
include: passing specific wavelengths of light from the light
source device of the spectrometer through the upper base material
in the body, or the assay site in which a reflective layer is
integrated; and detecting light reflected by the reflective layer
using the photodetector to measure light absorption of the specimen
in the assay site. The detection of the assay site using the
spectrometer may include measuring light absorption of the specimen
using the photodetector integrated in the body to obtain detection
results, and receiving the detection results using the wireless RF
IC integrated in the body to wirelessly transmit the results.
[0065] The cleaning process may further include adding a cleaning
solution to the assay site to clean the assay site. The cleaning
process may further include drying and dehydrating the assay site
based on centrifugal force caused by rotation of the body. The
remnant (debris) formed in the drying and dehydrating processes may
be trapped in the trash chamber based on centrifugal force.
[0066] In one embodiment, the body may include: a preparation
chamber to prepare DNA or RNA from blood serum obtained from the
specimen chamber; an amplification chamber to amplify the DNA and
RNA; and a fragmentation chamber to fragment the amplified DNA to a
predetermined length. For example, the DNA cut in a predetermined
size in the fragmentation chamber is incorporated into the assay
site in which DNA capture probes are arranged in an array and is
hybridized with the DNA capture probes having a complementary
sequence to produce double stranded DNA. Various embodiments to
detect hybridization are known in the art. The thin film centrifuge
device may further include, in addition to the chamber, a chamber
required for DNA amplification and fragmentation processes.
[0067] In one embodiment, the thin-film centrifuge device may
further include a thin film cylindrical magnet to perform
space-addressing of the amplification chamber or the fragmentation
chamber in the body.
[0068] In one embodiment, the thin-film centrifuge device may
further include a heating means to heat the amplification chamber
or the fragmentation chamber and a cooling means to cool the
same.
[0069] In one embodiment, the amplification chamber includes
performing a thermo cycle using polymerase chain reaction (PCR).
The space-addressing the amplification chamber or the fragmentation
chamber using the heating means may be carried out through radial
direction search and azimuthal direction search.
[0070] The analysis method may further include: isolating DNA or
RNA in the preparation chamber; fragmenting the amplified DNA to a
predetermined length; and labeling a marker on one terminal of
DNA.
[0071] The DNA amplification process may further include
rotation-cooling based on high-speed rotation of the body.
[0072] In one embodiment, the fragmentation process may further
include: incorporating DNAse in the amplification chamber after DNA
amplification; heating the DNAse using the heating means to
inactivate DNAse (incubation); and/or producing single stranded DNA
(denaturing).
BRIEF DESCRIPTION OF THE DRAWINGS
[0073] These and/or other aspects of the invention will become
apparent and more readily appreciated from the following
description of the embodiments, taken in conjunction with the
accompanying drawings of which:
[0074] FIGS. 1 and 2 are a sectional view and a plan view
illustrating a thin film layered centrifuge device and a thin film
layered centrifuge device drive to control operation of the device
according to one embodiment of the present invention;
[0075] FIG. 3 is a top view illustrating a slider provided with a
BOPM and a permanent magnet according to one embodiment of the
present invention;
[0076] FIG. 4 is a side view illustrating a thin film layered
centrifuge device drive to operate and control the thin film
centrifuge device of FIG. 1 according to one embodiment of the
present invention;
[0077] FIG. 5 illustrates a spectrometer using a grating mirror
according to one embodiment of the present invention;
[0078] FIGS. 6 to 8 illustrate a method for detecting the assay
site using a spectrometer on the thin film centrifuge device
according to one embodiment of the present invention;
[0079] FIGS. 9 and 10 illustrate a liquid valve to prevent liquid
leakage during centrifugation according to one embodiment of the
present invention;
[0080] FIG. 11 illustrates a stepwise centrifugation process;
[0081] FIG. 12 illustrates a bottle neck channel according to one
embodiment;
[0082] FIGS. 13 to 15 illustrate strips wherein various species of
tumor markers are fixed in the form of a line or spot on the porous
membrane according to one embodiment;
[0083] FIG. 16 illustrates a thin film centrifuge device wherein a
plurality of assay sites are arranged in parallel on different
sectors to perform a lab-on-a-chip process to assay various
specimens with respect to a single sample according to one
embodiment of the present invention;
[0084] FIG. 17 illustrates a stepwise process for transferring
blood serum from the specimen chamber to the buffer chamber by
alternately repeating the hydrophilic fluid flow process using the
liquid valve and the fluid flow process by centrifugal force;
[0085] FIG. 18 illustrates a state in which blood serum flows to
the assay site by centrifugal force as another embodiment different
from FIG. 17;
[0086] FIGS. 19 to 22 illustrate stepwise processes of the thin
film centrifuge device further comprising a dilute solution storage
chamber, as compared to the embodiment shown in FIG. 17; and
[0087] FIG. 23 illustrates a thin film layered centrifuge device
drive to front-load or top-load the thin film centrifuge device
according to one embodiment of the present invention.
DETAILED DESCRIPTION
[0088] Reference will now be made in detail to the embodiments of
the present invention, examples of which are illustrated in the
accompanying drawings, wherein like reference numerals refer to
like elements throughout.
[0089] Reference will now be made in detail to the embodiments of
the present invention, examples of which are illustrated in the
accompanying drawings, wherein like reference numerals refer to
like elements throughout.
[0090] FIGS. 1 and 2 are a sectional view and a plan view
illustrating a thin film layered centrifuge device and a thin film
layered centrifuge device drive to control operation of the device
according to one embodiment of the present invention.
[0091] Referring to FIGS. 1 and 2, the thin film centrifuge device
may be realized by integrating a lab-on-a-chip in thin film devices
such as conventional disc devices including CD-ROMs and DVDs. For
example, in one embodiment, provided are a thin film centrifuge
device 100 in which one or more chambers 130, 131a, 131b, 131c and
133 to store various buffer solutions required for analysis and to
perform various chemical processes and centrifugation, channels 92,
93 and 67 to enable the buffer solutions to be transferred, an
assay site 132 and a liquid valve 7 are integrated on a thin film
disc; and a thin film layered centrifuge device drive 100a to
control operation of the device.
[0092] As shown in FIG. 1, reference numeral 100 means a thin film
centrifuge device, which includes a body or a base material, formed
by laminating an upper base material 1, an intermediate base
material 2 and a lower base material 3. The thin film centrifuge
device also includes: the channels 92, 93 and 67 to allow fluids to
flow on the respective base materials during injection molding; a
liquid valve 7; a sample chamber 130; a specimen chamber 131a; a
remnant chamber 131b; an excess chamber 131c; an assay site 132 and
a trash chamber 133. These elements are closely adhered to one
another to constitute the thin film centrifuge device 100.
[0093] In one embodiment, the thin film centrifuge device may
further include an outlet 12 to discharge atmospheric pressure
generated by transferring samples from the specimen chamber 131a to
the sample chamber 130. The outlet 12 may be arranged opposite to
the direction of centrifugal force. In another embodiment, outlets
12 and 13 and a bottle neck channel 67 may be formed by a thin film
channel.
[0094] In one embodiment, the thin film channel may be formed
between the base materials 1, 2 and 3 by a channel-shaped thin-film
adhesive tape. That is, the base materials 1, 2 and 3 are adhered
to one another by a thin film adhesive tape to constitute the thin
film centrifuge device 100, and the thin film channels may be
provided in a region provided between the base materials in which
the thin film tape is omitted. The thickness of the thin film
channels may be determined depending on the thickness of the thin
film adhesive tape. Due to low thickness of the thin film channels,
strong capillary action to fluids may occur. In one embodiment, the
thickness of the thin film adhesive tape may be, for example, 0.001
mm to 0.1 mm.
[0095] Hereinafter, one embodiment wherein a sample is blood is
illustrated with reference to FIGS. 1 and 2.
[0096] Reference numeral 120 indicates a dispenser, a pipette, a
syringe, a lancet or a sample injection means to incorporate a
sample, reference numeral 121 indicates a sample inlet, reference
numeral 170 indicates a disc hole.
[0097] Reference numeral 130 indicates a sample chamber to store
blood injected from the sample inlet. Blood present in the sample
chamber 130 is transferred through the channel 92 to the specimen
chamber 131a and the remnant chamber 131b, while the body 100
initially rotates, and excess blood is transferred through
set-amount channels 93 to the excess chamber 131c. Then,
centrifugal force caused by rotation of the body 100 causes
independent centrifugation of blood stored in the specimen chamber
131a and the remnant chamber 131b, thus separating blood present in
the remnant chamber 131b as well as in the specimen chamber 131a
into blood serum and erythrocytes.
[0098] Reference numeral 67 indicates a bottle neck channel to
connect the specimen chamber 131a to the remnant chamber 131b. The
bottle neck channel 67 provides a passage, allowing transfer of the
erythrocytes present in the specimen chamber 131a to the remnant
chamber 131b and of the blood serum present in the remnant chamber
131b to the specimen chamber 131a, when bloods present in the
specimen chamber 131a and the remnant chamber 131b are centrifuged
by the centrifugal force generated by rotation of the body 100.
That is, the bottle neck channel 67 provides a passage, enabling
blood serum and erythrocyte to be freely moved from the specimen
chamber 131a to the remnant chamber 131b during centrifugation. As
shown in FIG. 1, the remnant chamber 131b is arranged close to the
external circumference of the body, as compared to the specimen
chamber 131a. For this reason, as a result of movement of blood
serum and erythrocyte through the bottle neck channel 67,
erythrocyte is collected in the remnant chamber 131b and blood
serum is collected in the specimen chamber 131a.
[0099] The bottle neck channel 67 may be composed of two thin film
channels for collection of blood serum and erythrocyte during
centrifugation. For such a bottle neck channel 67 composed of two
thin film channels, the remnant chamber 131b need not be provided
with an additional outlet. That is, the outlet 13 of the specimen
chamber 131a also acts as an outlet of the remnant chamber 131b by
the centrifugal force generated during rotation of the body 100.
However, the outlet 13 of the specimen chamber 131a cannot act as
an outlet of the remnant chamber 131b due to the absence of
centrifugal force when the body 100 is not rotated.
[0100] Although blood is centrifuged well, erythrocyte may be
admixed with blood serum again, when the body ceases rotation. That
is, in order to selectively extract only blood serum after
centrifugation, the body 100 should cease rotation. In this case,
erythrocyte is admixed with blood serum again, making it difficult
to extract only pure blood serum. In order to prevent this problem,
primarily, the specimen chamber 131a is isolated from the remnant
chamber 131b and secondarily, the bottle neck channel 67 is
interposed between the specimen chamber 131a and the remnant
chamber 131b to prevent fluid transfer therebetween, thirdly, the
remnant chamber 131b is provide in the form of a capillary tube
with a low height (narrow) to allow erythrocyte to remain in the
remnant chamber 131b due to inherent capillary action of the
remnant chamber 131b or bonding force between the surface of the
remnant chamber 131b and erythrocyte, thus preventing admixing of
the erythrocyte with blood serum in the specimen chamber 131a. The
bonding force between the surface of the remnant chamber 131b and
erythrocyte is derived from strong viscosity of erythrocyte. When
the remnant chamber 131b is in the form of a capillary tube,
centrifuged erythrocyte is not admixed with blood serum even when
the body is not rotated and remains on the surface of the remnant
chamber 131b. Accordingly, the blood serum in the specimen chamber
131a is not admixed with erythrocyte and maintains its state even
upon non-rotation of the body.
[0101] In one embodiment, the excess chamber 131c transfers
residual (excess) blood through the set-amount channel 93 to the
excess chamber 131c by centrifugal force generated by rotation of
the body 100. Depending on control of thickness of the set-amount
channel 93, the amount of blood (or blood serum) remaining in the
specimen chamber 131a may be determined. The blood having a height
higher than the thickness of the set-amount channel 93 may be
transferred through the set-amount channel 93 to the excess chamber
131c by centrifugal force generated by rotation of the body
100.
[0102] Reference numeral 290a is a base hole for alignment required
for producing and assembling the thin film centrifuge device 100.
The base hole 290a is injected into a fixture provided in a
jig.
[0103] Reference numeral 132 indicates an assay site in which a
capture probe for bonding (for example, biological specific
binding) to blood serum in the specimen chamber 131a is fixed,
and/or reagents for reactions (for example, biochemical reactions)
with specimens are stored.
[0104] Reference numeral 41 indicates a porous membrane or strip on
which the capture probe provided in the assay site 132 is fixed.
Reference numeral 13 is an outlet provided in the assay site 132,
which generates air stream upon rapid rotation of the body 100 to
promote drying of the strip 41. Prior to washing, the strip 41 is
dried to promote diffusion of a washing solution on the strip
during the washing and wash background noise-causing ingredients
due to diffusion force.
[0105] Blood serum trapped in the specimen chamber 131a during
centrifugation may be transferred through the liquid valve 7 to the
assay site 132 based on hydrophilic fluid flows when the body 100
ceases rotation.
[0106] Reference numeral 133 indicates a trash chamber to collect
debris generated by washing. That is, debris which does not bond to
the capture probe of the assay site 132 during rapid rotation of
the body 100 is trapped in the trash chamber 133 via the channel
94.
[0107] Reference numeral 211 indicates a slider equipped with a
permanent magnet 5a, which is connected to a slide motor 109 for
operation control.
[0108] The fluid flows are realized by the centrifugal force
derived from rotational force of the body, or super-hydrophilic
coatings of channels.
[0109] Reference numeral 291 indicates a thin-film cylindrical
magnet to spatially address the assay site 132.
[0110] Reference numeral 103a indicates an optical pick-up device
to read conventional optical discs (for example, CDs or DVDs),
reference numeral 103b is a detection device of the assay site 132
for quantitatively or qualitatively analyzing the assay site 132,
which may be a light transmission measurement apparatus, a
fluorescence detection device, an image sensor, a spectrophotometer
or a surface plasmon resonance (SPR) detection device, and the
optical pick-up device 103a and the assay site detection device
103b constitute a bio optical pickup module (BOPM) device 103.
Various embodiments of the fluorescence detection devices and SPR
detection devices are known in the art.
[0111] In one embodiment, the thin film centrifuge device for
space-addressing the assay site 132 includes the bio pickup optical
module (BOPM) device 103 equipped on the slider 211 and a slide
motor 109 to control movement of the BOPM device 103. A permanent
magnet 5a is mounted on the slider 211, to attract the thin film
cylindrical magnet 291, and movement of coordinates of BOPM device
can be controlled by control of the slide motor 109. The space
addressing to the assay site 132 may be realized by radial and
azimuthal direction search.
[0112] One embodiment of the radial and azimuthal direction search
is as follows. The radial direction search is a process for
transferring the permanent magnet 5a in a radial direction, which
is carried out by moving the permanent magnet 5a on the slider 211
on the corresponding diameter of the thin film cylindrical magnet
291. Then, the azimuthal direction search is required to overlap
the permanent magnet 5a and the thin film cylindrical magnet 291 on
the corresponding diameter. The azimuthal direction search is
carried out by slowly rotating a spindle motor 102 or repeatedly
operating short rotation and stop of the same, while the slider 211
is stopped. During slow rotation or several short rotation of the
spindle motor, the permanent magnet 5a on the slider 211
corresponds to the thin-film cylindrical magnet 291 present on the
corresponding diameter, the body 100 cannot be rotated by slow or
short rotation due to strong attraction force therebetween. In this
case, the permanent magnet 5a and the thin film cylindrical magnet
291 are aligned.
[0113] In addition, in another embodiment, the azimuthal direction
search may be carried out by controlling rotation of stepper motors
which are mechanically connected to the shaft of the spindle motor
102 in need of the azimuthal direction search. The rotation of the
stepper motors enables control of the rotation angle of the spindle
motor 102.
[0114] Reference numeral 116b indicates a flexible cable to connect
control signals required for BOPM 103 on the slider 211, which is
connected to a central control device 101 through a wafer or a
harness 116a.
[0115] Reference numeral 181 is a turn table on which the thin film
centrifuge device 100 is placed, and the thin film centrifuge
device 100 is placed in front of or on the top of the turn table
through a central hole 170 of the body. Reference numeral 188
indicates a memory-provided wireless RF IC or an electric tag
device, which includes protocols for lab-on-a-chip processes,
detection results of the assay site 132, analysis algorithm,
standard control values for detection, and location information of
the assay site 132, information associated with bioinformatics, and
information associated with self diagnosis. In addition, the
memory-provided wireless RF IC or an electric tag device may store
personal privacy encryption information, and identification (ID) of
the thin film centrifuge device, thus preventing others' use
without permission. The wireless RF IC 188 includes a smart IC
card. The information of the wireless RF IC 188 is supplied to the
central control device 101 through wireless transmission/reception
and may thus be utilized in personal privacy encryption. Reference
numeral 110 indicates a wireless electric wave generator to supply
power to the wireless RF IC 188. An alternating magnetic field
generated by the wireless electric wave generator 110 senses an
induction coil provided in the wireless RF IC 188 in accordance
with Fleming's rule, to generate a sufficient amount of electricity
and supply the same to the wireless RF IC 188. The wireless
electric wave generator is provided with a multipole permanent
magnet to generate electricity on the induction coil provided in
the wireless RF IC 188 based on an alternating magnetic field
generated by rotation of the body 100. In one embodiment, the
multipole permanent magnet may be circumferentially arranged on a
tray to load the body 100.
[0116] For the thin film centrifuge device according to one
embodiment, the wireless RF IC 188 performs temperature measurement
to measure the temperature of the assay site 132 and
wirelessly-transmits the same to the central control device 101.
The assay site 132 can maintain a constant temperature using a
heating or cooling apparatus, when the temperature is excessively
high or low. In one embodiment, the temperature of the assay site
132 is within the range of 30 to 37 degrees in the reaction with
specimens, for example, in which biochemical activity and stability
are considered.
[0117] For the thin-film centrifuge device according to one
embodiment, the wireless RF IC 188 may include information such as
test date and test results according to residual pesticide and
antibiotic tests of the thin film centrifuge device, efficient
periods, agriculture and stockbreeding regions, production and
farming (culturing) history, distribution history, contact
information of farmers, price and organic products. Consumers and
agriculture and stockbreeding distribution enterprises may purchase
crops and livestock products with ease using the information.
General consumers can obtain information by touching the thin film
centrifuge device 100 to the RF IC detector, or loading the same on
the thin film centrifuge drive 100a.
[0118] For the thin film centrifuge device according to one
embodiment, the wireless RF IC 188 can store the test results of
the thin film centrifuge device in a memory provided therein.
[0119] For the thin film centrifuge device according to one
embodiment, the wireless RF IC 188 controls the detection device of
the assay site, and wirelessly transmits the results thus obtained
to the central control device 101 or a storage device 112 or an
input/output device 111.
[0120] For the thin film centrifuge device according to one
embodiment, the input/output device may be a universal serial bus
(USB) or IEEE1394 or ATAPI or SCSI or a device having a
communication standard of internet network. In addition, user
information, such as height, weight, gender and age, of the thin
film centrifuge device 100 can be input through the input/output
device 111.
[0121] FIG. 2 illustrates an absorption pump provided between the
terminal of the liquid valve 7 and the assay site 132 with a sample
pad 41a or an absorption pad 41b to transfer blood serum in the
specimen chamber 131a to the assay site 132 by an absorption force
to absorb blood serum in the specimen chamber 131a through a U- or
V-shaped hydrophilic channel 7 according to one embodiment. The
specimen chamber 131a can be emptied by transferring blood serum in
the specimen chamber 131a to the assay site 132 via the absorption
pump. After specimens present in the specimen chamber 131a are
ejected through the liquid valve 7 to the assay site 132, fluids in
the remnant chamber 131b do not transfer to the liquid valve 7 due
to strong capillary action of the bottle neck channel 67. That is,
strong capillary action of the bottle neck channel 67 to fluids is
equivalent to the fluid movement force of the absorption pump and
the fluid does not transfer to the assay site 132. Reference
numeral 41b indicates an absorption pad, reference numeral 41a
indicates a sample pad and a conjugate pad, and these pads are
connected to the terminals of a porous membrane 41c.
[0122] FIG. 3 is a top view illustrating a slider provided with a
BOPM 103 and a permanent magnet 5a according to one embodiment of
the present invention. The movement of the slider can be controlled
by worm gear connections 109a and 109b connected to the shaft of a
slide motor 109. The slider may slide using slide arms 108a and
108b as guides. The slide arms 108a and 108b are engaged through
screws 110a, 110b, 110c and 110d on the body of the thin film
layered centrifuge device (100a, shown in FIG. 1). Reference
numeral 116b indicates a flexible cable, which is connected through
a wafer or a harness 116a. Reference numeral 181 indicates a turn
table rotated by the spindle motor (102, shown in FIG. 1).
[0123] FIG. 4 is a side view illustrating a film layered centrifuge
device drive 100a to operate and control the thin film centrifuge
device 100 of FIG. 1 according to one embodiment of the present
invention. Reference numeral 300 indicates a body to support the
thin film layered centrifuge device drive 100a. A circuit substrate
140 is continuously engaged in the body 300 of the thin film
layered centrifuge device drive under the thin film layered
centrifuge device drive, and the central control device 101, the
storage device 112 and the input/output device 111 to control the
thin film layered centrifuge device drive 100a are arranged on the
circuit substrate 140. The central control device 101 controls a
spindle motor 102 to rotate and brake the thin film centrifuge
device 100, controls movement of the bio optical pickup module
(BOPM) arranged on the slider 211 by control of the slide motor
109, and moves the permanent magnet 5a for space-addressing the
assay site 132 of the thin film centrifuge device 100. The
permanent magnet 5a can efficiently transfer an electric field to a
thin film-type cylindrical magnet (291, see FIG. 1). In addition,
the central control device 101 decides whether the disc loaded on
the thin film layered centrifuge device drive 100a is a
conventional optical disc (for example, music CDs, CD-Rs, and game
CDs/DVDs) or the thin film centrifuge device 100. When the disc is
a conventional optical disc, the device 101 transfers information
read from the disc from the optical pick-up device (103a, see FIG.
3) to the storage device 112 or input/output device 111, transfers
data to be written to the optical pick-up device (103a, see FIG.
3), or performs conventional operations for optical discs, for
example, supplies various control signals required for reading and
writing to respective elements.
[0124] In one embodiment, upon loading of the thin film centrifuge
device, inherent ID of the thin film centrifuge device 100 is
wirelessly transmitted to the central control device 101 through
the wireless RF IC 188 on the thin film layered centrifuge device
to inform the central control device 101 of the fact that the disc
loaded on the thin film layered centrifuge device drive 100a is a
thin film centrifuge device.
[0125] In one embodiment, detection results associated with the
assay site 132 are transferred via wireless communication through
the wireless RF IC 188 provided on the thin film centrifuge device
100 to the central control device 101, the storage device 112 or
the input/output device 111. The detection associated with the
assay site 132 may be carried out by transferring image information
associated with the assay site 132 obtained by an image sensor 144
arranged on the circuit substrate 140 to the central control device
101 or storage device 112 or input/output device 111. Reference
numeral 104 is a compression device of the thin film centrifuge
device 100 loaded on a disc hole, which performs compression by
means of attraction force, based on a magnetic field with a turn
table 181 and is designed such that vertical movement and no-load
rotation are allowed.
[0126] Reference numeral 144a indicates one or more light-emitting
diodes (LEDs) to illuminate the image sensor, the image sensor 144
or the LED 144a is mounted on the slider 211 or equipped on or
under the assay site 132. In one embodiment, the LEDs include
multicolor LEDs to emit various wavelengths of light, which can
obtain reaction intensity of the assay site 132 as image
information represented by color intensity under illumination of
various wavelengths, and enables quantitative or qualitative
analysis of response results of the assay site 132 based on
2-dimensional correlation between the wavelengths and color
intensity. The multicolor LEDs include R, G and B LEDs. Reference
numeral 107 indicates a laser beam generator used to excite
fluorescently labeled specimens in the assay site. In this case,
the image sensor 144 enables image information associated with the
assay site to be obtained. Reference numeral 108 indicates a
spectrophotometer, which outputs a plurality of light wavelengths
to measure light transmission or light absorption of the assay site
and measures light transmission or light absorption of respective
wavelengths to detect reaction results of the assay site 132. A
spectrometer generally includes a light source, a wavelength
selector, a specimen vessel (test tube or assay site 132), and a
photodetector, which is known in the art. A spectrometer measures
light absorbance of the specimen solution of the assay site, after
it is calibrated using a blank solution such that light
transmission is 100% (light absorbance of 0%). A light source
should uniformly emit a sufficient energy of light required for
specimen analysis. Examples of the light source that can be used in
the present invention include tungsten filament lamps, hydrogen or
deuterium lamps, white light LEDs and lasers. In one embodiment,
the light source may be an LD module in which white light LEDs or
RGB lasers or a plurality of laser diodes (LDs) are integrated. The
RGB laser is a module device comprising 3 lasers to emit red, green
and blue light. Combining various powers of these lasers enables
generation of various wavelengths of light required for specimen
analysis. The LD module is a module of a plurality of laser diodes
(LDs) having different wavelengths. In accordance with the LD
module, light absorbance of the specimen at the corresponding
wavelengths of light are measured, while LDs to output the
corresponding wavelength of light are sequentially turned on. For
the spectrometer, it is important to obtain specific wavelength of
radiation from the light emitted from the light source.
Monochromatic radiation is ideal, but in practice, this is very
difficult. Accordingly, for light showing a predetermined range of
wavelength distribution, monochromatization level can be
represented by specifying the spectrum band width. Sensitivity and
resolution of measurement are proportional to the closeness of the
light emitted from the light source to a single wavelength. The
desired wavelength of light can be obtained using a wavelength
selector and the wavelength selector may be a filter or a grating
mirror or a combination thereof. The grating mirror acts as a prism
to distribute and reflect incident light at various
wavelengths.
[0127] FIG. 5 illustrates a spectrometer (108, See FIG. 4) using a
grating mirror according to one embodiment of the present
invention.
[0128] As shown in FIG. 5, white light emitted from a light source
40 passes through a lens 42, converges into a beam, and passes
through a primary H-slit and a V-slit 45a to generate a spot beam.
When the spot beam is incident on a grating mirror 43, light
reflected from the grating mirror 43 is separated into different
wavelengths in a topological space. In order to collect specific
wavelengths from the light reflected from the grating mirror 43 and
separated in the topological space, a secondary H-slit and V-slit
45b is set at a specific angle. In this case, the wavelengths of
light passing through the secondary H-slit and V-slit 45b can be
varied by rotating the grating mirror 43. That is, the desired
specific wavelengths of light can be obtained by controlling the
rotation angle of the grating mirror 43.
[0129] After the specific wavelengths of light thus obtained pass
through the assay site 132, the photodetector 46 measures light
absorption, light transmission or color intensity of the specimen
in the analysis site to perform qualitative or quantitative
analysis of reaction results of the site specimen. Methods for
qualitatively or quantitatively analyzing the reaction results of
the specimen include an end point, rate assay, initial rate
methods, or the like, which are known in the art.
[0130] Reference numeral 40 indicates a light source of the
spectrometer 108 and the wavelength selector includes a stepper
motor 44 to control the rotation angle of the grating mirror 43, a
lens 42 to converge light generated from the light source and the
primary H-slit and V-slit 45a to convert the converged beam into a
spot beam, the grating mirror 43 to separate the spot beam into
various wavelengths, and the secondary H-slit and V-slit 45b to
pass only a specific angle of beam (that is, specific wavelengths
of light) reflected from the grating mirror 43. The specific
wavelengths of light obtained by the light source 40 and the
wavelength selector pass through the assay site 132 and light
absorption of the specimen present in the assay site is measured
with the photodetector 46, to qualitatively or quantitatively
analyze reaction results of the specimen. Light absorption of the
specimen in the assay site can be measured at various wavelengths
by passing various wavelengths of light through the assay site 132,
while rotating the stepper motor 44.
[0131] In one embodiment, an optical fiber may be used instead of
the primary H-slit and V-slit or secondary H-slit and V-slit.
[0132] In one embodiment, hereinafter, one of various combinations
of the light source, the lens, the primary H-slit and V-slit or the
primary optical fiber 45a, the grating mirror 43, the secondary
H-slit and V-slit 45b or secondary optical fiber will be referred
to as a light source device 99a. The LD module and the RGB laser
module may singly constitute the light source device 99a. In this
case, the light source device 99a may be simplified.
[0133] FIGS. 6 to 8 illustrate a method for detecting the assay
site 132 using a spectrometer 108 on the thin film centrifuge
device 100 according to one embodiment of the present invention.
Reference numeral 555 indicates a transparent opening to detect the
photodetector 46.
[0134] As shown in FIG. 6, the photodetector 46 of the spectrometer
108 is arranged on the thin film centrifuge device 100 and the
light source device 99a is arranged thereunder. A plurality of
assay sites 132 arranged in a circumferential direction in the thin
film centrifuge device 100 are detected using the spectrometer 108
in which the light source device 99a and the photodetector 46 are
modulated. In this case, as the thin film centrifuge device 100
rotates, one to one correspondence occurs between the spectrometer
108 and each of the assay sites 132 provided in a circumferential
direction in the thin film centrifuge device 100, thus realizing
space addressing and detection. The measurement of light absorbance
of the specimen solution in the plurality of assay sites using the
spectrometer 108 is performed, after the device is calibrated using
a blank solution such that light transmission reaches 100% (light
absorbance: 0). In one embodiment, one or more of the plurality of
assay sites may include a blank solution chamber for
calibration.
[0135] As can be seen from the left image of FIG. 7, a reflective
layer 99b is integrated in an upper base material 1 or an assay
site in the thin film centrifuge device 100, and the spectrometer
108 in which the light source device 99a and the photodetector 46
are modulated in a lower side of the thin film centrifuge device
100 is arranged. The specific wavelengths of light obtained from
the light source device 99a pass through the assay site 132 and the
photodetector 46 measures light reflected from the reflective layer
99b, to measure light absorption of the specimen in the assay
site.
[0136] The right image of FIG. 7 shows a case in which the
photodetectors 46 are integrated in the assay sites 132 of the thin
film centrifuge device 100. In this case, the photodetectors 46 are
arranged such that they correspond one-to-one to the plurality of
assay sites 132. When the photodetectors 46 are integrated in the
thin film centrifuge device 100, the optical path is shortened,
reception sensitivity of the photodetectors 46 increases and
sensitivity is improved. The detection results of the
photodetectors 46 integrated in the thin film centrifuge device 100
are read by the wireless RF IC 188 and are then wirelessly
transmitted to the central control device (101, see FIG. 1).
[0137] As shown in FIG. 8, the reflective layer 99b illustrated in
the left image of FIG. 7 is integrated in an upper base material 1,
and a plurality of assay sites (132, see FIG. 7) are arranged in a
circumferential direction of the thin film centrifuge device 100.
The spectrometer 108 one-to-one corresponds to each of the assay
sites provided in a circumferential direction in the thin film
centrifuge device 100 to realize sequential detection by
space-addressing. In this case, the light source device 99a emits
wavelengths of light suitable for the specimen in the respective
assay sites 132, to measure light absorbance. In one embodiment,
space-addressing of the assay site by radial direction search and
azimuthal direction search performed by mounting the spectrometers
108 on the slider 211 may be performed prior to sequential
detection of the assay sites 132 using the spectrometers 108. The
image sensor includes a CCD, a CMOS and a line image sensor for
sensing light amount in pixel units. In one embodiment, the line
image sensor includes a linear sensor array or a contact image
sensor (CIS). In one embodiment, the BOPM 103 including the image
sensor can move the slider 211 to obtain image information of the
assay site. Prior to detection of the assay site, space-addressing
of the assay site by radial direction search and azimuthal
direction search performed by mounting the spectrometers 108 on the
slider 211 may be performed.
[0138] FIGS. 9 and 10 illustrate a liquid valve to prevent liquid
leakage during centrifugation according to one embodiment of the
present invention. The liquid valve 7 prevents transfer of blood
serum to the assay site 132 through a V- or U-shaped channel 7 upon
high-speed rotation of the body 100. In addition, FIGS. 9 and 10
are detailed views of the liquid valve 7. The liquid valve 7 is
broadly divided into two parts, i.e., an inward channel 7a and an
outward channel 7b. The inward channel 7a refers to a channel
extended toward the center of the body (opposite to a direction of
the centrifugal force) and the outward channel 7b refers to a
channel extended toward the centrifugal force direction. The
operation of the liquid valve 7 is as follows. When the body 100
rotates at a high speed, the liquid leaked from the specimen
chamber 131a is primarily charged in the inward channel 7a. Once
the leaked liquid fills the inward channel 7a, centrifugal force
acts in a radial direction toward the liquid contained in the
inward channel 7a, thus preventing further leakage of the liquid
from the specimen chamber 131a. Otherwise, the leaked liquid is
withdrawn to the specimen chamber 131a by centrifugal force. That
is, when liquid is leaked from the specimen chamber 131a upon
high-speed rotation of the body 100, further leakage of the liquid
can be prevented due to the equivalence between the force to
further leak the liquid from the specimen chamber 131a and
centrifugal force inherently acting on the already leaked liquid.
This prevention of the leakage of liquid based on the centrifugal
force acting on the leaked liquid is referred to as a liquid valve
operation in one embodiment of the present invention.
[0139] In one embodiment, the specimen chamber 131a may further be
provided at an outlet thereof with a liquid valve to prevent liquid
leakage during centrifugation.
[0140] In one embodiment, the liquid valve includes liquid valves
realized by a V- or U-shaped channel or a super-hydrophilic coated
channel to operate the liquid valve.
[0141] FIG. 11 is an image showing the specimen chamber 131a and
the remnant chamber 131b of the thin film centrifuge device 100 of
FIG. 2, for illustration of a centrifugation process.
[0142] FIG. 11 illustrates a stepwise process in which the blood
transferred from the sample chamber 130 to the specimen chamber
131a and the remnant chamber 131b by rotation of the body 100 is
separated into blood serum and erythrocyte by centrifugation. In
step 1, blood is transferred from the sample chamber 130 to the
specimen chamber 131a and the remnant chamber 131b during initial
rotation of the body and blood having a height higher than the
height of a set-amount channel 93 is moved to the excess chamber
131c by centrifugal force. In addition, blood cannot be moved
through the liquid valve 7 to the assay site 132 and is thus
retained in the specimen chamber 131a. Step 2 is an intermediate
state of centrifugation, in which blood present in the specimen
chamber 131a and blood present in the remnant chamber 131b are
independently centrifuged, based on centrifugal force caused by
rotation of the body and is thus separated into blood serum and
erythrocyte. The centrifugal force caused by rotation of the body
induces independent centrifugation of blood in the specimen chamber
131a and the remnant chamber 131b and allows erythrocyte in the
specimen chamber 131a to be moved through the bottle neck channel
67 to the remnant chamber 131b. In addition, blood serum
centrifuged in the remnant chamber 131b is moved through the bottle
neck channel 67 into the specimen chamber 131a. That is, the bottle
neck channel 67 provides a passage, allowing blood serum and
erythrocyte separated during centrifugation to be smoothly moved
from the specimen chamber 131a to the remnant chamber 131b. The
remnant chamber 131b is arranged closer to the circumference than
the specimen chamber 131a. For this reason, as a result of
centrifugation, erythrocyte is collected in the remnant chamber
131b and blood serum is collected in the specimen chamber 131. Step
3 shows, as a result of Step 2, a state in which erythrocyte is
collected in the remnant chamber 131b and blood serum is collected
in the specimen chamber 131 after centrifugation. In step 4, the
blood serum of the specimen chamber 131 hydrophilic-flows through
the liquid valve 7 to the assay site 132, when the body 10 does not
rotate after centrifugation. In step 5, only a predetermined amount
of blood serum in the specimen chamber 131a is moved to the assay
site 132. That is, only the predetermined amount of blood serum is
moved to the assay site 132 and fluids present in the bottle neck
channel 67 and the remnant chamber 131b are not moved to the assay
site 132 and remain therein. The amount of blood serum moved to the
assay site 132 is determined by the amount of blood serum stored in
the specimen chamber 131a.
[0143] Such a phenomenon is due to the following five causes.
[0144] The bottle neck channel 67 is used as the thin film channel,
strong capillary action occurs when the body is not rotated, the
fluids present in the remnant chamber 131b are not moved through
the bottle neck channel 67 to the specimen chamber 131a.
Accordingly, transfer of erythrocyte from the remnant chamber 131b
to the specimen chamber 131a can be prevented. The remnant chamber
131b is provided in the form of a capillary tube chamber and
erythrocyte stored therein cannot be thus readily ejected. The
bonding force between the surface of the remnant chamber 131b and
the erythrocyte inhibits easy escape of erythrocyte from the
remnant chamber 131b. When the body 100 does not rotate, the bottle
neck channel 67 is clogged by viscose blood serum, thus making
movement of erythrocyte stored in the remnant chamber 131b
difficult. The remnant chamber 131b has no outlet, thus making
movement of erythrocyte stored therein difficult.
[0145] FIG. 12 illustrates a bottle neck channel 67 according to
one embodiment. This detailed view illustrates a cross-section of
the bottle neck channel 67, based on a base line to join reference
numerals 67a and 67b.
[0146] The bottle neck channel 67 is composed of two thin film
channels, which are formed by a first thin film adhesive tape 1a to
join an upper base material 1 to an intermediate base material 2
and a second thin film adhesive tape 2a to join an intermediate
base material 2 to a lower base material 3. The bottle neck channel
67 by formed by these two thin film channels provides a passage,
enabling easy transfer of erythrocyte from the specimen chamber
131a to the remnant chamber 131b, or of blood serum from the
remnant chamber 131b to the specimen chamber 131a during
centrifugation. That is, the bottle neck channel 67 serves as a
bottle neck channel to prevent fluid transfer upon non-rotation of
the body and as a passage for blood serum and erythrocyte during
centrifugation.
[0147] FIGS. 13 to 15 illustrate strips wherein various species of
tumor markers are fixed in the form of a line or spot on the porous
membrane according to one embodiment. Hereinafter, each of various
species of tumor marker lines or spots is referred to as a test
line.
[0148] Reference numeral 41a is a conjugate pad, a sample pad, or a
combination thereof and reference numeral 41b is an absorbent pad.
Reference numeral 41c is a porous membrane. A gold conjugate, an
enzyme linked antibody or a label such as a fluorescent marker may
be deposited in a lyophilized form on the conjugate pad. The
capture probe (for example, capture antibody) can fix the tumor
markers. The tumor markers may be selected from the group
consisting of AFP, PSA, CEA, CA19-9, CA125 and A15-3. The capture
antibody can fix glutamine synthetase (GS), a specific marker for
Alzheimer's. The capture can fix myocardial infarction antibody
markers such as myoglobin, CK-MB and Troponin I (TnI).
[0149] In one embodiment, the test line wherein one or more markers
or capture probes for AIDS, myocardial infarction, residual
antibiotics, residual pesticides, allergy and breast cancer tests
are fixed on the porous membrane 41c may be applied to response
tests using immunochromatography. Immunochromatography is a test
method wherein immunochemistry is combined with chromatographic
assay, which utilizes specific immune-response of antibody to
antigens, color rendering and flowability of colloidal gold
particles, and transfer of molecules on the porous membrane by
capillary phenomenon. Immunochromatography is a convenient and
rapid one-step method to perform processes including sample
dilution, cleaning and color-rendering (chromogenesis) based on
reaction of enzyme-complex with a substrate involved in
conventional multi-step immunoanalysis methods. In addition, this
method can easily detect test results without using any specific
apparatus, thus having advantages of ease, high economic efficiency
and rapid detection of test results. The capture antibody can fix,
in addition to tumor markers, antibodies for reference and control
lines. Plural reference lines may be present. The reaction
concentration of the reference line may be a cutoff value to enable
easy detection of negative or positive responses. For example, the
cutoff value of the reference line may be selected from 3 ng/ml, 4
ng/ml, 10 ng/ml, 20 ng/ml, 30 ng/ml, 40 ng/ml and 50 ng/ml.
[0150] In one embodiment, the test line comprises qualitative or
quantitative analysis based on the difference in reaction intensity
between the reference line and the test line.
[0151] In one embodiment, the test line comprises qualitative or
quantitative analysis based on the difference in reaction intensity
between the background and the test line.
[0152] In one embodiment, the test line comprises qualitative or
quantitative analysis performed by determining reaction intensity
of the test line through a linear function of reaction intensity
formed by a plurality of reference lines.
[0153] In one embodiment, the test line comprises qualitative or
quantitative analysis performed by determining reaction intensity
of the test line through a linear function of reaction intensity
formed by the reference line and the control line.
[0154] In one embodiment, antibodies to capture free PSA are
immobilized on the reference line and antibodies to capture total
PSA are immobilized on the test line, to calculate percent free PSA
(fPSA %). fPSA(%) can be calculated as a ratio of total PSA to free
PSA. Details of the total PSA and free PSA are known in the art. In
addition, free PSA may be immobilized on the test line and the
total PSA may be immobilized on the reference line. In another
embodiment, antibodies to capture free PSA are immobilized on the
reference line and antibodies to capture proPSA are immobilized on
the test line, to calculate percent proPSA (% proPSA). The proPSA
(%) can be obtained by a ratio of free PSA to proPSA. Details of
the proPSA are known in the art. Alternatively, proPSA may be
immobilized on the reference line and the free PSA may be
immobilized on the test line. Also, % fPSA and % proPSA can be
simultaneously calculated by immobilizing free PSA, proPSA and
total PSA on one porous membrane.
[0155] In one embodiment, the reaction intensity can be obtained
from image information represented by color intensity thereof under
illumination using various wavelengths of LEDs. The reaction
results of the assay site 132 can be quantitatively or
qualitatively analyzed based on the quadratic relationship between
the various wavelengths and color intensities. The reference line
shows a positive response when the specimen is diffused into the
absorption pad 41b and may be used for validity of tests using
strips. The test results are considered efficient, when the
reference line is positive. The porous membrane 41c may be used in
a flow through or lateral flow manner, which is known in the art. A
specimen or a cleaning solution may be injected into the sample pad
41a. The flow through-type porous membrane may utilize strips
wherein various tumor markers, disease markers or antibodies are
immobilized on the porous membrane 41c. When the specimen is
injected into the sample pad 41a, the specimen absorbed by the
sample pad 41a is diffused throughout the porous membrane 41c by
the capillary phenomenon and is thus biochemically specifically
bonded to the capture antibody. The absorption pad 41b for
promoting diffusion may be arranged on the terminal of the porous
membrane 41c. Also, a conjugate pad may be connected to the sample
pad. In this case, the liquid specimen injected into the sample pad
is linked to a gold conjugate, an enzyme-linked antibody or a
fluorescent material on the conjugate pad and a complex thus
obtained is diffused into the porous membrane 41c. When a cleaning
solution is injected into the sample pad 41a, the cleaning solution
absorbed on the sample pad 41a is diffused into the porous membrane
41c by capillary action to clean materials not bound to the capture
antibody or non-specifically bound thereto and thereby remove
background noise from the porous membrane 41c.
[0156] In one embodiment, the assay site 132 may be set by
connecting the strip 41 to the terminal of the liquid valve 7 and a
portion of the sample pad 41a.
[0157] In one embodiment, the detection of the assay site 132 using
the image sensor 144 comprises treating the upper base material 1
with a non-transparent material or coating the same with a
non-transparent paint to prevent light scattering by illumination
and noise caused by damage to substrates. In this case, for
example, the transparency of the upper base material may be 20 to
50%.
[0158] FIG. 16 illustrates a thin film centrifuge device wherein a
plurality of assay sites 132 are arranged in parallel on different
sectors to perform assay of various specimens with respect to a
single sample, for example, processes required for a lab-on-a-chip
to assay biochemical reactions according to one embodiment of the
present invention.
[0159] The term "biochemical reaction assay" used herein includes
for example, assay of GOT, GPT, ALP, LDH, GGT, CPK, amylase,
T-protein, albumin, glucose, T-cholesterol, triglycerides,
T-bilirubin, D-bilirubin, BUN, creatinine, I. phosphorus, calcium,
and uric acid in blood.
[0160] Reference numerals 132a, 132b, 132c and 132d are assay sites
acting as chambers for the biochemical reaction, which store
specimens to analyze and diagnose the biochemical reaction and
results thereof and to perform biochemical reactions with blood
serum supplied from the specimen chamber 131a. Reference numeral 7
indicates a liquid valve to prevent leakage of liquids during
centrifugation of blood.
[0161] Reference numeral 290a indicates a base hole, and reference
numeral 131c indicates an excess chamber. Reference numerals 154a,
154b, 154c and 154d indicate thin film valves. Reference numerals
13a, 13b, 13c, 13d and 14 indicate outlets.
[0162] During rotation of the body 100, blood stored in the sample
chamber 130 is centrifuged. As a result, blood serum is stored in
the specimen chamber 131a and erythrocyte is stored in the remnant
chamber 131b. Set-amount chambers 140a, 140b, 140c and 140d are
chambers to supply a predetermined amount of specimens to the
corresponding assay sites 132a, 132b, 132c and 132d, and the volume
of the set-amount chambers determines an amount of specimens
supplied to the corresponding assay site. The liquid valve 7 and a
concentric channel 9 are coated with a super-hydrophilic material
and an overflow chamber 132e is coated with a hydrophobic material.
Accordingly, when the body 100 does not rotate, the blood serum in
the specimen chamber 131a hydrophilically flows through the liquid
valve 7 through the concentric channel 9. The set-amount chambers
140a, 140b, 140c and 140d are chambers coated with a
super-hydrophilic material, which are filled with blood serum,
while specimens are passed through the concentric channel 9. In
this case, the overflow chamber 132e is hydrophobic and thus only
the concentric channel 9 and the set-amount chambers 140a, 140b,
140c and 140d are filled with the specimens. The concentric channel
9 is designed such that it has a concentric circle and thus
receives homogeneous centrifugal force during rotation.
Accordingly, after the concentric channel 9 is filled with the
specimen and the body 100 rotates again, specimens are stored in
the set-amount chambers 140a, 140b, 140c and 140d and remain
therein, and the specimens filling the concentric channel 9 are
discharged through the overflow chamber 132e by centrifugal force.
Then, the thin film valves 154a, 154b, 154c and 154d open to allow
specimens to flow from the set-amount chambers 140a, 140b, 140c and
40d to the respective assay sites 132a, 132b, 132c and 132d and
thus induce biochemical reactions of the specimens with reagents.
In one embodiment, the thin film valves 154a, 154b, 154c and 154d
are concentrically arranged and thus simultaneously open. Then,
biochemical reaction results of the specimens can be qualitatively
or quantitatively analyzed by measuring light absorption of
specimens in the assay sites 132a, 132b, 132c and 132d using the
spectrometer.
[0163] In one embodiment, when the concentric channel 9 is designed
such that it has a concentric circle and thus receive identical
centrifugal force to rotate the thin film centrifuge device 100,
only specimens present in the set-amount chambers 140a, 140b, 140c
and 140d are stored and remain therein, but the specimens filling
the concentric channel 9 overcome the hydrophobic barrier formed in
the overflow chamber 132e by centrifugal force and are discharged
to the overflow chamber 132e.
[0164] In one embodiment, the thin film valves 154a, 154b, 154c and
154d, for example, include valves such as burst valves formed in a
thin-film shape to open/close openings using open/close means of
thin film valves, such as valve open/close using micro-beads (or
thin-film cylindrical (circular) magnets) provided in a hole with a
movable permanent magnet or electric magnet arranged in an upper or
lower part of the body; valve open/close by mechanical force; valve
open/close by centrifugal force; valve open/close by dissolution
and solidification based on chemical actions; open/close by shape
memory alloys restored to original shape by heat or chemicals;
valve open/close using air drops generated by electrolysis; valve
open/close using air drops generated by heat; valve open/close by
thermal expansion and contraction of micro-beads; valve open/close
by electrostatic force; valve open/close by magnetic force; valve
open/close by laser heat; valve open/close using a temperature
gradient; valve open/close of an actuator based on ultrasonic
waves; valve open/close based on a pump or physical pressure; valve
open/close by micro particles expanded or contracted by super-high
frequency; capillary burst valves; hydrophobic burst valves; valve
open/close by magnetic fluid; and valve open/close by thermal
expansion and contraction of air.
[0165] The hydrophobic burst valve uses a fluid flow barrier formed
on the interface between the hydrophilic channel and the
hydrophobic chamber, and includes hydrophobic burst valves wherein
fluids cannot flow under centrifugal force of a cutoff value or
less, but fluids overcome a fluid flow barrier under centrifugal
force of a cutoff value or higher and move to the hydrophobic
chamber. The fluid flow barrier is formed due to the facts that
hydrophilic fluids cannot readily pass through a hydrophobic
chamber and that the hydrophilic channel traps fluids based on
inherent capillary action of the fluids.
[0166] The assay sites 132a, 132b, 132c and 132d are hydrophobic
chambers and the thin film valves 154a, 154b, 154c and 154d use the
hydrophobic burst valve. In this case, the set-amount chambers
140a, 140b, 140c and 140d are chambers coated with a
super-hydrophilic material, and can form a fluid flow barrier at
the interface with the assay site. Thin film valves including the
burst valve are known in the art.
[0167] In another embodiment, a thin film valve may be further
provided between an inward channel 7a of the liquid valve 7 and an
outlet of the specimen chamber 131a shown in FIG. 2. In this case,
when the thin film valve closes even upon non-rotation of the body
100, fluids in the specimen chamber 131a do not flow to the assay
site 132, and after the thin film valve opens, fluids
hydrophilic-flow through the liquid valve 7 to the assay site
132.
[0168] In one embodiment, azimuthal assay site search for measuring
the spectrometer 108 can be carried out by controlling the rotation
angle of the thin film centrifuge device using the stepper motors
or gears connected thereto.
[0169] In one embodiment, azimuthal assay site search for
measurement via the spectrometer 108 can be carried out through the
azimuthal valve search process performed by arranging the thin film
cylindrical magnet for searching assay sites on the circumference
of the body, or by space-addressing assay sites using a blank
solution chamber during rotation of the body 100 to sequentially
measure light absorption in the respective assay sites during
rotation of the body. In this case, the body further includes a
black solution chamber having a diameter equivalent to the assay
site to store the blank solution. The light absorbance of specimens
in the respective assay sites is measured, after the spectrometer
is calibrated such that light transmission of the blank solution is
100% (light absorbance: 0). The light absorbance of the blank
solution is always zero, thus enabling sensing of the blank
solution chamber during rotation of the body and thus realizing
space-addressing of the assay sites based on the blank solution
chamber.
[0170] The embodiments may also be applied to thin film centrifuge
devices for processes associated with a lab-on-a-chip to perform
enzyme-linked immunosorbent assays (ELISAs) or chemical
luminescence immunosorbent assays (CLISAs). Various embodiments
thereof are known in the art.
[0171] The embodiments may be also applied to thin film centrifuge
devices for processes associated with a lab-on-a-chip for residual
pesticide assays and residual antibiotic assays. In this case, test
results derived from detection results are displayed on a computer
monitor, history is reported to the servers of the corresponding
government offices or food enterprises through automatic or manual
remote-access thereto using an internet protocol based network, or
test results and histories are stored in a memory of wireless RF
ICs (electric tags). The corresponding government office can detect
residual pesticide and the food enterprises can obtain purchase
information of fresh crops and livestock products. In addition, the
corresponding government office links this information on a web, to
provide information for purchasing fresh crops and livestock
products for consumers by direct transaction. Enzymes and markers
for assaying residual pesticides contain enzymes and markers for
assaying pesticides contained in enzymes, vegetables or fruits, for
example, the most-generally used organophosphorus and carbamate
insecticides. The enzyme may include acetylcholinesterase (AChE).
Various embodiments thereof are known in the art.
[0172] FIG. 17 illustrates an assay site 132 different from that of
FIG. 2 as another embodiment of the thin film centrifuge device
100.
[0173] In this case, the thin film centrifuge device further
comprises a plurality of assay sites 132a, 132b and 132c to provide
biochemical reaction assay or immunological analysis using the
strip 41; a liquid valve 7 to temporarily store the specimens of
the specimen chamber 131a to retain blood serum in the specimen
chamber 131a during rotation of the body, when the body does not
rotate, and to provide a hydrophilic fluid flow passage to transfer
blood serum from the specimen chamber 131a to the buffer chamber
131d; thin film valves 155a, 155b and 155c to independently supply
blood serum in the transferred buffer chamber 131d to a plurality
of assay sites; and a hydrophilic channel 8 to transfer blood serum
through hydrophilic fluid flows from the buffer chamber 131d to the
corresponding assay site, when the thin film valves 155a, 155b and
155c open. Based on these elements, the thin film centrifuge device
enables multiplex assay of a single sample. In this case, transfer
of blood serum from the specimen chamber 131a to the buffer chamber
131d is carried out by alternately repeating a hydrophilic fluid
flow process through the liquid valve 7 and a fluid flow process by
centrifugal force.
[0174] FIG. 17 illustrates a stepwise process for transferring
blood serum from the specimen chamber 131a to the buffer chamber
131d by alternately repeating the hydrophilic fluid flow process
using the liquid valve 7 and the fluid flow process by centrifugal
force.
[0175] In step 1, during rotation of the body after centrifugation,
blood serum is collected in the specimen chamber 131a and
erythrocyte is collected in the remnant chamber 131b.
[0176] In step 2, when the body does not rotate after
centrifugation, blood serum of the specimen chamber 131a is
charged, through the liquid valve 7, into the inward channel 7a and
the outward channel 7b and hydrophilic-flows to the buffer chamber
131d.
[0177] In step 3, blood serum in the outward channel 7b flows in
the buffer chamber 131d based on centrifugal force caused by
rotation of the body.
[0178] In step 4, when the body ceases rotation, blood serum from
the specimen chamber 131a is charged in the inward channel 7a and
the outward channel 7b again through the liquid valve 7 and then
hydrophilic-flows in the buffer chamber 131d.
[0179] In step 5, by repetition of steps 3 and 4, the total blood
serum gradually flows from the specimen chamber 131a to the buffer
chamber 131d.
[0180] In step 6, when the thin film valve 155a opens, blood serum
flows through the hydrophilic channel 8 from the buffer chamber
131d to the corresponding assay site 132a. The buffer chamber 131d
of FIG. 17 may be coated with a super-hydrophilic material. In this
case, blood serum can readily flow from the specimen chamber to the
buffer chamber by operation of the absorption pump.
[0181] FIG. 18 illustrates a state in which blood serum flows to
the assay site by centrifugal force as another embodiment different
from FIG. 17. In this case, the thin film valves 155a, 155b and
155c may be a hydrophobic burst valve or a capillary tube burst
valve.
[0182] The thin film valves 155a, 155b and 155c are hydrophobic
burst valves or capillary tube burst valves formed by a fluid flow
barrier formed on the interface between the hydrophilic channel 8
coated with a hydrophilic material, and the hydrophobic chamber,
the assay site 132a, 132b or 132c. Such a fluid flow barrier allows
blood serum to not flow at a centrifugal force less than a
predetermined cutoff value and allows blood serum to overcome the
fluid flow barrier and then flow to the assay site 132a, 132b or
132c at a centrifugal force of the predetermined cutoff value or
higher. In this case, the centrifugal force of step 3 may be
applied in a level lower than the centrifugal force to overcome the
fluid flow barrier.
[0183] FIGS. 19 to 22 illustrate stepwise processes of the thin
film centrifuge device further comprising a dilute solution storage
chamber, as compared to the embodiment shown in FIG. 17. Reference
numeral 131e indicates a dilute solution storage chamber to store a
dilute solution.
[0184] FIGS. 19 and 20 illustrate a phenomenon in which the dilute
solution stored in a dilute solution storage chamber 131e flows to
a buffer chamber 131f upon opening of the burst valve 150, while
specimens in the specimen chamber 131a are centrifuged. In this
case, the dilute solution stored in the buffer chamber 131f is
retained by the liquid valve 11. Similarly, specimens in the
specimen chamber 131a are also retained in the specimen chamber
131a through the liquid valve 7 during centrifugation. Excess
dilute solution in the buffer chamber 131f flows through a
set-amount channel 10 to an excess chamber 131g to allow a desired
amount of the dilute solution to be stored in the buffer chamber
131f.
[0185] FIG. 21 illustrates a process in which blood serum in the
specimen chamber 131a is charged in the inward channel 7a and the
outward channel 7b through the liquid valve 7, when the body ceases
rotation, and hydrophilic-flows to a mixing chamber 131h. In
addition, FIG. 21 illustrates a process in which the dilute
solution in the buffer chamber 131f is charged in an inward channel
11a and an outward channel 11b through the liquid valve 11 and then
hydrophilic-flows to the mixing chamber 131h.
[0186] FIG. 22 illustrates a phenomenon wherein the specimen and
the dilute solution are gradually transferred to the mixing chamber
131h through repetition of a process for transferring fluids in the
outward channels 7b and 11 b to the mixing chamber 131h based on
centrifugal force caused by rotation of the body and a process for
filling the inward channels 7a and 11a and the outward channels 7b
and 11 b with hydrophilic fluids, after ceasing rotation of the
body. Accordingly, the dilute solution is mixed with the specimens
in the mixing chamber 131h to produce a diluted specimen.
[0187] In the process for gradually transferring the specimen and
the dilute solution to the mixing chamber 131h via repetition of
the hydrophilic fluid flow through the liquid valve and fluid flow
based on centrifugal force, the specimen is gradually mixed with
the dilute solution to maximize mixing efficiency between these
fluids. Hereinafter, the mixing of the specimen with the dilute
solution is referred to as a gradual mixing during the process for
gradually transferring the specimen and the dilute solution to the
mixing chamber 131h.
[0188] Closing strength of the hydraulic burst valve is determined,
based on an adhesion area of a thin-film adhesive tape, when holes
are clogged by the thin film adhesive tape, and a valve to allow
the thin film adhesive tape to be detached at a disc rotational
speed (centrifugal force) or higher to overcome the closing
strength and thereby open the hole.
[0189] The burst valve may be, for example, a hydraulic burst
valve, which is known in the art.
[0190] FIG. 23 illustrates a thin film layered centrifuge device
drive 100a to front-load or top-load the thin film centrifuge
device according to one embodiment of the present invention. The
thin film centrifuge device 100 may be loaded on the thin film
layered centrifuge device drive 100a. Reference numeral 751
indicates a case of the thin film layered centrifuge device drive
and reference numeral 750a indicates a tray to front-load the thin
film centrifuge device 100. In addition, reference numeral 750b
indicates a cover to perform top-loading. When the cover opens, the
hole 170 of the thin film centrifuge device may be mounted on the
turn table. Based on a loading type, one may be selected from
reference numeral 750a and reference numeral 750b. In addition,
according to one embodiment of the present invention, the thin film
layered centrifuge device drive may optionally comprise a
reading/searching button 745 or a stop button 746 to read
conventional optical discs. Reference numeral 744 indicates an
on/off button of the thin film centrifuge device drive.
[0191] Reference numeral 760 indicates a display device to show a
process state and mode of the thin film layered centrifuge device
drive. A light-emitting diode or an LCD may be used as the display
device. The display device 760 displays whether the loaded disc is
a thin film centrifuge device or an optical disc, analysis results
or a main process state of the thin film layered centrifuge device
drive. Alternatively, the display device 760 displays a graphical
user interface and a process rate in accordance with the step in
the form of a percent (%), a bar graph or a pie graph.
[0192] Reference numeral 111 indicates an input/output device to
realize automatic or manual remote contact with a doctor of the
corresponding field through an internet network and to
remote-transmit diagnosis results and questionnaires to the doctor,
if necessary. Then, patients wait for a prescription from the
doctor.
[0193] The thin film layered centrifuge device drive shown in FIG.
8 may further comprise a speaker, a moving-image camera and/or a
microphone. In the case of non-advanced cancers, a level of tumor
markers in blood does not increase, and in the case of early
cancers, a level of tumor markers in blood is within a normal range
and it increases and positive ratio thereof also increases, as the
tumors progress. In one embodiment, taking this fact into
consideration, the centrifuge device comprises statistical software
to historically manage detection results derived from quantitative
analysis of the assay site to provide information associated with
regular trace diagnosis to a user.
[0194] In addition, in one embodiment, the thin film layered
centrifuge device drive may further comprise software to read and
analyze the response results and to analyze negative or positive
response, presence of risk groups or values.
[0195] Also, in one embodiment, the thin film layered centrifuge
device drive allows side loading or vertical loading of the thin
film centrifuge device.
[0196] As apparent from the afore-going, the thin film layered
centrifuge device and an analysis method using the same according
to one embodiment may be applied to thin-film devices to diagnose
and detect a small amount of biological and/or chemical materials
in fluids, such as a lab-on-a-chip, protein chips and DNA chips.
For example, the thin film centrifuge device may be applied to
integrate centrifuge devices in thin film discs such as
conventional CD-ROMs or DVDs.
[0197] Although a few embodiments of the present invention have
been shown and described, it would be appreciated by those skilled
in the art that changes may be made in these embodiments without
departing from the principles and spirit of the invention, the
scope of which is defined in the claims and their equivalents.
* * * * *