U.S. patent application number 13/677462 was filed with the patent office on 2013-06-06 for reagent splitting/dispensing method based on reagent dispensing nozzle and reagent splitting/dispensing mechanism.
The applicant listed for this patent is Noe Miyashita, Hideyuki Noda, Masahiro Okanojo. Invention is credited to Noe Miyashita, Hideyuki Noda, Masahiro Okanojo.
Application Number | 20130139898 13/677462 |
Document ID | / |
Family ID | 43836797 |
Filed Date | 2013-06-06 |
United States Patent
Application |
20130139898 |
Kind Code |
A1 |
Miyashita; Noe ; et
al. |
June 6, 2013 |
REAGENT SPLITTING/DISPENSING METHOD BASED ON REAGENT DISPENSING
NOZZLE AND REAGENT SPLITTING/DISPENSING MECHANISM
Abstract
A reagent splitting/dispensing method based on a reagent
dispensing nozzle controls a split or dispensation amount of
reagent, includes a waiting process of disposing a first air layer
between an interface of the operation fluid and a nozzle tip end in
the reagent dispensing nozzle; a first moving process of moving the
reagent dispensing nozzle to a position above the reagent to be
split; a second moving process of depositing the nozzle tip end in
the reagent; an air layer adjusting process of increasing the
occupation amount of the operation fluid in the reagent dispensing
nozzle and decreasing the occupation amount of the first air layer,
between the first moving process and the second moving process; and
a reagent splitting process of decreasing the occupation amount of
the operation fluid in the reagent dispensing nozzle and filling
the reagent into the reagent dispensing nozzle from the nozzle tip
end.
Inventors: |
Miyashita; Noe; (Tokyo,
JP) ; Noda; Hideyuki; (Tokyo, JP) ; Okanojo;
Masahiro; (Tokyo, JP) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Miyashita; Noe
Noda; Hideyuki
Okanojo; Masahiro |
Tokyo
Tokyo
Tokyo |
|
JP
JP
JP |
|
|
Family ID: |
43836797 |
Appl. No.: |
13/677462 |
Filed: |
November 15, 2012 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
13009037 |
Jan 19, 2011 |
8333936 |
|
|
13677462 |
|
|
|
|
Current U.S.
Class: |
137/2 ;
422/509 |
Current CPC
Class: |
Y10T 436/118339
20150115; G01N 35/1002 20130101; G01N 2035/102 20130101; Y10T
436/2575 20150115; B01L 3/52 20130101; G01N 35/1011 20130101; G01N
2035/1032 20130101; F17D 3/00 20130101; Y10T 137/0324 20150401;
Y10T 436/119163 20150115; G01N 21/763 20130101 |
Class at
Publication: |
137/2 ;
422/509 |
International
Class: |
B01L 3/00 20060101
B01L003/00; F17D 3/00 20060101 F17D003/00 |
Foreign Application Data
Date |
Code |
Application Number |
Jan 19, 2010 |
JP |
2010-009037 |
Claims
1. A reagent splitting/dispensing method based on a reagent
dispensing nozzle that controls a split amount or a dispensation
amount of a reagent by an operation fluid disposed in the reagent
dispensing nozzle, the reagent splitting/dispensing method
comprising: a waiting process of disposing a first air layer
between an interface of the operation fluid and a nozzle tip end in
the reagent dispensing nozzle; a first moving process of moving the
reagent dispensing nozzle to a position above the reagent to be
split; a second moving process of depositing the nozzle tip end in
the reagent; an air layer adjusting process of increasing the
occupation amount of the operation fluid in the reagent dispensing
nozzle and decreasing the occupation amount of the first air layer,
between the first moving process and the second moving process; a
reagent splitting process of decreasing the occupation amount of
the operation fluid in the reagent dispensing nozzle and filling
the reagent into the reagent dispensing nozzle from the nozzle tip
end.
2. The reagent splitting/dispensing mechanism comprising: a
triaxial actuator in which a movement axis in a horizontal
direction is set to an X axis and a Y axis and a movement axis in a
vertical direction is set to a Z axis; a reagent dispensing nozzle
which is moved by the triaxial actuator; a pump unit which is
connected to the reagent dispensing nozzle and controls an
operation fluid disposed in the reagent dispensing nozzle; and a
control unit which outputs a first air layer arrangement signal to
decrease the occupation amount of the operation fluid in the nozzle
and dispose a first air layer between an interface of the operation
fluid and a nozzle tip end in the reagent dispensing nozzle to the
pump unit, outputs a first movement signal to move the reagent
dispensing nozzle to a position above the reagent to be split and a
second movement signal to deposit the nozzle tip end in the reagent
to the triaxial actuator after the first air layer is disposed in
the reagent dispensing nozzle, outputs an air layer adjustment
signal to increase the occupation amount of the operation fluid in
the reagent dispensing nozzle to the pump unit, between the output
of the first movement signal and the output of the second movement
signal, outputs a reagent split signal to decrease the occupation
amount of the operation fluid in the nozzle and fill the reagent
into the reagent dispensing nozzle from the nozzle tip end to the
pump unit after the nozzle tip end is deposited in the reagent.
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application is a continuation application of U.S.
application Ser. No. 13/009,037, filed Jan. 19, 2011, the contents
of which are incorporated herein by reference.
BACKGROUND
[0002] 1. Field of the Invention
[0003] The present invention relates to a mechanism that
splits/dispenses a trace amount of a liquid (micro order), and more
particularly, to a method that splits/dispenses a reagent by a
reagent dispensing nozzle in a luminescence measuring device and a
mechanism thereof.
[0004] 2. Description of the Related Art
[0005] In various clinical medicine sites, food factories,
medicinal drug manufacturing factories, and basic research sites,
an aseptic work environment and predetermined biological
cleanliness are required. In an environment where the biological
cleanliness is required, the number of microorganisms (the number
of viable bacteria) in the air (floating bacteria in the air), the
number of falling bacteria, and the number of adhesive bacteria are
measured. As a method that measures the number of floating bacteria
in the air, it is general to use a sampler of the floating bacteria
in the air to collect the floating bacteria by natural falling of
the floating bacteria or sucking air of a constant amount in
collecting the floating bacteria.
[0006] In this method, the floating bacteria are collected on an
agar plate and are cultured by an incubator for two or three days,
and the number of colonies generated after culturing is used as the
number of bacteria. However, in this method, a long time is needed
to culture the viable bacteria.
[0007] Meanwhile, as a method that enables measurement of the
number of microorganisms in a short time, a method that measures
adenosine triphosphate (ATP) corresponding to a component in a cell
by a bioluminescence method and converts the number of
microorganisms is known.
[0008] In the bioluminescence method, a luciferin-luciferase
luminescence reaction is used, the ATP amount is calculated from
the luminescence amount of light generated by mixing and reacting a
luminescence reagent containing substrate luciferin and enzyme
luciferase and a sample solution containing the ATP extracted from
the cells of the microorganisms, and the number of viable bacteria
is calculated on the basis of the ATP amount per viable bacterium.
In Japanese Patent Application Laid-Open (JP-A) No. 11-155597, a
kit that measures the number of viable bacteria using the
luminescence reaction is disclosed.
[0009] According to the method that measures the number of viable
bacteria using the kit disclosed in JP-A No. 11-155597, a
measurement time can be decreased. However, when the viable
bacteria of the minute amount are measured, the luminescence amount
becomes the minute amount. For this reason, background luminescence
is greatly affected by mixing of the remaining ATP or the ATP other
than the measurement object, and superior measurement precision
cannot be obtained.
[0010] Meanwhile, in JP-A No. 2008-249628, a luminescence measuring
device that can suppress background luminescence due to viable
bacteria adhered to a nozzle to dispense a reagent or a remaining
ATP and can quickly measure luminescence with high precision is
disclosed.
[0011] According to the luminescence measuring device that is
disclosed in JP-A No. 2008-249628, even in luminescence measurement
where the viable bacteria of the minute amount are measured, it is
assumed that the viable bacteria of the minute amount can be
quickly measured with high precision. However, when the viable
bacteria of the minute amount are measured by the luminescence
measuring device, a measurement value may be greatly affected by
contamination in the device.
SUMMARY OF THE INVENTION
[0012] Accordingly, it is an object of the present invention to
provide a splitting/dispensing method based on a reagent dispensing
nozzle and a reagent splitting/dispensing mechanism that can
prevent contamination of an operation fluid in the reagent
dispensing nozzle to split/dispense a reagent when the reagent
dispensing nozzle is in a waiting state or generation of cross
contamination due to falling of a droplet when the reagent
dispensing nozzle is moved, and split/dispense the reagent with
high precision.
[0013] To solve the aforementioned problem to be solved by the
invention, the reagent splitting/dispensing method based on a
reagent dispensing nozzle that controls a split amount or a
dispensation amount of a reagent by an operation fluid disposed in
the reagent dispensing nozzle, the reagent splitting/dispensing
method is featured by including the following: a waiting process of
disposing a first air layer between an interface of the operation
fluid and a nozzle tip end in the reagent dispensing nozzle; a
first moving process of moving the reagent dispensing nozzle to the
position right above the reagent becoming a split object; a second
moving process of depositing the nozzle tip end in the reagent; a
reagent splitting process of decreasing the occupation amount of
the operation fluid in the reagent dispensing nozzle and filling
the reagent into the reagent dispensing nozzle from the nozzle tip
end; a third moving process of evacuating the tip end of the
reagent dispensing nozzle from the reagent; a reagent protecting
process of disposing a second air layer between an interface of the
split reagent and the nozzle tip end; a fourth moving process of
moving the reagent dispensing nozzle to the reagent dispensation
position, after the reagent protecting process; a reagent
dispensing process of increasing the occupation amount of the
operation fluid in the reagent dispensing nozzle and ejecting the
split reagent; an operation fluid protecting process of disposing
the first air layer between the interface of the operation fluid
and the nozzle tip end, after dispensing the reagent; and a fifth
moving process of evacuating the reagent dispensing nozzle to the
waiting position, in a state where the first air layer is disposed
between the interface of the operation fluid and the nozzle tip
end.
[0014] Also, the reagent splitting/dispensing method having the
aforementioned characteristic features preferably includes the
following steps: an air layer adjusting process of increasing the
occupation amount of the operation fluid in the reagent dispensing
nozzle and decreasing the occupation amount of the first air layer,
between the first moving process and the second moving process. By
using these processes, when the reagent is filled into the reagent
dispensing nozzle, the capacity of the first air layer that is
disposed between the reagent and the interface of the operation
fluid can be decreased. Thereby, the splitting/dispensing error of
the reagent due to the compression or expansion of the air layer
can be decreased.
[0015] Also, the reagent splitting/dispensing method having the
aforementioned characteristic may includes the following: a
temporary evacuating process of moving the reagent dispensing
nozzle to the operation fluid discharge position after the reagent
dispensing process; and an operation fluid ejecting process of
ejecting a part of the operation fluid from the nozzle tip end, at
the operation fluid discharge position. By using these processes,
the operation fluid that exists near the interface where the
possibility of the reagent being mixed or contaminated is high can
be discharged. Thereby, generation of the cross contamination of
the reagent or the contamination of the reagent can be
prevented.
[0016] Further, in the reagent splitting/dispensing method having
the aforementioned characteristics, it is preferable that, in the
reagent dispensing process, the process proceeds to the temporary
evacuating process, after the first air layer remains. By adopting
these units, the reagent that is filled into the reagent dispensing
nozzle can be completely ejected, and the operation fluid in the
reagent filled nozzle can be prevented from being ejected.
[0017] Also, in order to solve the aforementioned problems to be
solved by the invention, the reagent splitting/dispensing mechanism
according to an exemplary embodiment of the invention includes a
triaxial actuator in which a movement axis in a horizontal
direction is set to an X axis and a Y axis and a movement axis in a
vertical direction is set to a Z axis, a reagent dispensing nozzle
which is moved by the triaxial actuator, and a pump unit which is
connected to the reagent dispensing nozzle an controls an operation
fluid disposed in the reagent dispensing nozzle. The reagent
splitting/dispensing mechanism is featured by including: a control
unit which outputs a first air layer arrangement signal to decrease
the occupation amount of the operation fluid in the nozzle and
dispose a first air layer between an interface of the operation
fluid and a nozzle tip end in the reagent dispensing nozzle to the
pump unit, outputs a first movement signal to move the reagent
dispensing nozzle to the position right above the reagent becoming
a split object and a second movement signal to deposit the nozzle
tip end in the reagent to the triaxial actuator after the first air
layer is disposed in the reagent dispensing nozzle, outputs a
reagent split signal to decrease the occupation amount of the
operation fluid in the nozzle and fill the reagent into the reagent
dispensing nozzle from the nozzle tip end to the pump unit after
the nozzle tip end is deposited in the reagent, outputs a third
movement signal to evacuate the tip end of the reagent dispensing
nozzle from the reagent to the triaxial actuator, outputs a reagent
protection signal to dispose a second air layer between an
interface of the split reagent and the nozzle tip end to the pump
unit, outputs a fourth movement signal to move the reagent
dispensing nozzle to the reagent dispensation position to the
triaxial actuator, after the second air layer is disposed in the
reagent dispensing nozzle, outputs a reagent dispensation signal to
increase the occupation amount of the operation fluid in the
reagent dispensing nozzle and eject the reagent to the pump unit,
after the reagent dispensing nozzle reaches the reagent
dispensation position, outputs an operation fluid protection signal
to decrease the occupation amount of the operation fluid in the
reagent dispensing nozzle and dispose the first air layer between
the interface of the operation fluid and the nozzle tip end to the
pump unit, after the reagent is ejected from the reagent dispensing
nozzle, and outputs a fifth movement signal to evacuate the reagent
dispensing nozzle where the first air layer is disposed after the
reagent is dispensed to the waiting position to the triaxial
actuator.
[0018] Also, in the reagent splitting/dispensing mechanism having
the aforementioned characteristic features, the control unit
outputs an air layer adjustment signal to increase the occupation
amount of the operation fluid in the reagent dispensing nozzle to
the pump unit, between the output of the first movement signal and
the output of the second movement signal. By using the control unit
having the above configuration, when the reagent is filled into the
reagent dispensing nozzle, the capacity of the first air layer that
is disposed between the reagent and the interface of the operation
fluid can be decreased. Thereby, the splitting/dispensing error of
the reagent due to the compression or expansion of the air layer
can be decreased.
[0019] In the reagent splitting/dispensing mechanism having the
aforementioned characteristic features, it is preferable that the
control unit outputs a temporary evacuation signal to move the
reagent dispensing nozzle to the operation fluid discharge position
to the triaxial actuator, after the reagent dispensation signal is
output, and the control unit outputs an operation fluid ejection
signal to increase the occupation amount of the operation fluid in
the reagent dispensing nozzle to eject a part of the operation
fluid from the tip end of the reagent dispensing nozzle moved to
the operation fluid discharge position, to the pump unit. By using
the control unit having the above configuration, the operation
fluid that exists near the interface where the possibility of the
reagent being mixed or contaminated is high can be discharged.
Thereby, generation of the cross contamination of the reagent in
the reagent dispensing nozzle or the contamination of the reagent
can be prevented.
[0020] In the reagent splitting/dispensing mechanism having the
aforementioned characteristic features, the reagent dispensation
signal is a signal to control the occupation amount of the
operation fluid to completely eject the reagent filled into the
reagent dispensing nozzle and eject a part of the first air layer.
By adopting these units, the reagent that is filled into the
reagent dispensing nozzle can be completely ejected, and the
operation fluid in the reagent filled nozzle can be prevented from
being ejected.
[0021] In the reagent splitting/dispensing mechanism that has the
above characteristics, the pump unit is preferably configured as a
syringe pump. By using this configuration, the reagent can be
split/dispensed with high precision.
[0022] According to the reagent splitting/dispensing method by use
of a reagent splitting nozzle having the above characteristics,
when the reagent dispensing nozzle to split/dispense the reagent is
in awaiting state, contamination of the operation fluid in the
reagent dispensing nozzle and falling of the droplet can be
prevented. Thereby, generation of the cross contamination can be
prevented and the reagent can be split/dispensed with high
precision.
[0023] According to the reagent splitting/dispensing mechanism
having the above characteristics, the method is executed and an
effect based on the method can be achieved.
BRIEF DESCRIPTION OF THE DRAWINGS
[0024] FIG. 1 is a block diagram showing the configuration of a
luminescence measuring device;
[0025] FIG. 2 is a schematic view showing the lateral configuration
of a measuring unit;
[0026] FIG. 3A is a front block diagram showing a relationship
between the schematic configuration of a triaxial actuator and a
reagent dispensing nozzle;
[0027] FIG. 3B is a top block diagram showing a relationship
between the schematic configuration of a triaxial actuator and a
reagent dispensing nozzle;
[0028] FIG. 4 is a reference perspective view showing a
relationship between a Z-axis mechanism unit, a fixed block, and
the reagent dispensing nozzle;
[0029] FIG. 5A is a top view showing the configuration of a
reagent/carrier container mounting unit;
[0030] FIG. 5B is a top view showing a reagent cartridge;
[0031] FIG. 6 is a flow view showing an aspect of luminescence
measurement based on a luminescence measuring device;
[0032] FIGS. 7A to 7G are diagrams showing a state where the
reagent is split/dispensed using the reagent splitting/dispensing
mechanism according to an exemplary embodiment of the invention;
and
[0033] FIG. 8 is a flowchart showing each operation process in the
reagent splitting/dispensing mechanism.
DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0034] Hereinafter, an embodiment of a reagent splitting/dispensing
method based on a reagent dispensing nozzle and a reagent
splitting/dispensing mechanism according to the present invention
will be described in detail with reference to the drawings.
[0035] First, the entire configuration of a luminescence measuring
device (biomedical device) 10 that mounts a reagent
splitting/dispensing mechanism according to this embodiment will be
described with reference to FIG. 1. The luminescence measuring
device 10 that is described in this embodiment includes a measuring
unit 12 and a collecting unit 80.
[0036] The measuring unit 12 has a reagent dispensing unit 14, a
hot water supply unit 42, a reagent/carrier container mounting unit
54, a buffer supply unit 64, a filter unit 72, a photomultiplier
tube (PMT) unit 78, and an input/control unit (hereinafter, simply
called control unit 11). These components are disposed in an outer
shell.
[0037] The reagent dispensing unit 14 is configured using a
triaxial actuator 16, a reagent dispensing nozzle 24, and a syringe
pump (pump unit) 32 as a basic body. The triaxial actuator 16 is a
unit to move the reagent dispensing nozzle to be described in
detail below to the desired position. For this reason, the triaxial
actuator 16 includes a Y-axis mechanism unit 18, an X-axis
mechanism unit 20, and a Z-axis mechanism unit 22 to be described
in detail below. The Y-axis mechanism unit 18 is disposed on a
device that is rarely spatially restricted. For this reason, in the
measuring unit 12 according to this embodiment, a stepping motor
18a is used as a driving actuator and a movable unit 18c that is
attached to a linear guide 18b is slid by a driving belt 18d.
[0038] Meanwhile, the X-axis mechanism unit 20 and the Z-axis
mechanism unit 22 attached to the movable unit 18c are difficult to
have a spatial margin. For this reason, in the X-axis mechanism
unit 20 and the Z-axis mechanism unit 22, a compact actuator is
adopted. The compact actuator is a small actuator that is
configured by incorporating a thrust axis system having a large
diameter in a hollow rotor and integrating a motor and a protrusion
shaft with each other. As a driving principle, a driving system is
set as the stepping motor and the protrusion shaft is set as a ball
screw. For this reason, positioning with high precision is realized
while a size is decreased.
[0039] The reagent dispensing nozzle 24 is a nozzle that
splits/dispenses various reagents used in luminescence measurement
by the desired amount. As shown in FIGS. 3A, 3B, and 4, the reagent
dispensing nozzle 24 is supported by a fixing block 28 that is
included in a slide guide 26 attached to the compact actuator
corresponding to the Z-axis mechanism unit 22. By adopting this
support form, an elevating operation can be stabilized.
[0040] FIG. 3A is a front block diagram showing a relationship
between the schematic configuration of the triaxial actuator 16 and
the reagent dispensing nozzle 24. FIG. 3B is a block diagram
showing the configuration of a top surface in FIG. 3A. FIG. 4 is a
reference perspective view showing a relationship between the
Z-axis mechanism unit 22 and the reagent dispensing nozzle 24.
[0041] To a rear end of the reagent dispensing nozzle 24, a
dispensing operation tube 30 that is connected to the syringe pump
32 to be described in detail below is connected. The reagent
dispensing nozzle 24 splits the reagent by applying the negative
pressure to an inner part of the nozzle through the dispensing
operation tube 30 and dispenses the split reagent by applying the
positive pressure to the inner part of the nozzle. The reagent
dispensing nozzle 24 may be composed of a resin or metallic tube,
in addition to a glass tube.
[0042] The syringe pump 32 performs a control operation of an
operation fluid (pure water in this embodiment) to split/dispense
the reagent by the reagent dispensing nozzle 24. The syringe pump
32 is configured using a syringe 34, a plunger 36, and an actuator
38 as a basic body. The syringe 34 is a tank that stores the pure
water corresponding to the operation fluid. The plunger 36 is a
pushing rod that applies the negative pressure or the positive
pressure to the inner part of the syringe 34 to introduce the pure
water into the syringe 34 and discharge the pure water from the
syringe 34. The actuator 38 is a driving unit that pushes in or
pulls out the plunger 36. If the stepping motor and the ball screw
are used in the actuator 38, position control with high precision
is enabled.
[0043] To a tip end of the syringe 34 in the syringe pump 32 having
the above configuration, one end of the dispensing operation tube
30 is connected. The other end of the dispensing operation tube 30
is connected to the reagent dispensing nozzle 24 described above.
By connecting the dispensing operation tube 30 in the
above-described way, the pure water is collected by pulling out the
plunger 36 and the reagent is injected (split) into the reagent
dispensing nozzle 24. In contrast, when the plunger 36 is pushed
therein, the power water that is discharged from the inner part of
the syringe 34 is moved to the reagent dispensing nozzle 24. For
this reason, the pressure in the reagent dispensing nozzle 24
increases and the reagent that is collected in the reagent
dispensing nozzle 24 is ejected (dispensed).
[0044] To the dispensing operation tube 30, the buffer supply tube
70 that is connected to the buffer supply unit 64 to be described
in detail below is connected through a distribution valve 40 such
as a three-way valve. By this configuration, the pure water that is
the operation fluid collected in the dispensing operation tube 30
can be regularly switched. Thereby, an error of measurement data
can be suppressed from being generated due to contamination of the
operation fluid.
[0045] The hot water supply unit 42 supplies pure water to dilute a
collection carrier. The hot water supply unit 42 is configured
using a peristaltic pump 44, a heater 46, and a hot water supply
nozzle 48 as a basic body. The peristaltic pump 44 is configured
using a resin tube, a resin roller, and an actuator as a basic body
(none of them are shown in the drawings). The resin tube is a tube
that is used to send a solution and a conveyance fluid (pure water
in this embodiment) flows through the resin tube. The resin tube is
preferably configured to have flexibility and durability, because
the resin tube may be crushed by a roller. For example, the resin
tube may be composed of a silicon tube. The roller repeats the
rotation and the revolution while crushing the resin tube and
extrudes the conveyance fluid closed in a crushing region to a
revolution direction of the roller. In the resin tube that is
crushed by the roller, the power that causes a shape of the resin
tube to return to an original shape works. Since the conveyance
fluid is a non-compression fluid, even though plural rollers
continuously revolve and extrude the conveyance fluid, the
operation is continuously performed. Any actuator that can rotate
the plural rollers may be used.
[0046] According to the peristaltic pump 44 having the above
configuration, since a place contacting the conveyance fluid (pure
water in this embodiment) is only an inner part of the tube where
the conveyance fluid flows, the bump is not contaminated. For this
reason, an aseptic state can be easily maintained and cleaning can
be easily performed.
[0047] The heater 46 heats the pure water that is the conveyance
fluid. The configuration of the heater 46 is not particularly
limited. However, when the heater 46 needs to be configured to have
a small size, a cartridge heater or a tube heater is preferably
adopted. For example, when the cartridge heater is adopted, a
metallic tube 46b may be wounded around the outer circumference of
a heater body 46a and the pure water corresponding to the
conveyance fluid may be circulated in the wound metallic tube 46b.
This is because the pure water in the metallic tube 46b is heated
by heat transmission, if the above configuration is used. When the
tube heater is adopted, a rubber heater is wound around the resin
tube (tube) and the conveyance fluid that is circulated in the
resin tube is heated. In this configuration, if the silicon resin
is used in the resin tube, a heat transfer coefficient is
increased. Since the resin tube and the rubber heat are configured
to have the flexibility, a degree of freedom of arrangement is high
and a heated region can be secured to be long. For this reason, the
temperature after heating can be avoided from being lowered, that
is, the temperature can be stabilized. The arrangement position of
the heater 46 is not particularly limited. However, it is
preferable to decrease the solution sending distance after the
heating to prevent the temperature after the heating from being
lowered. Therefore, in the measuring unit 12 according to this
embodiment, the heater 46 is disposed between the peristaltic pump
44 and the hot water supply nozzle 48 to be described in detail
below.
[0048] The hot water supply nozzle 48 is an ejection nozzle that
supplies the hot water (pure water), which is sent by the
peristaltic pump 44 and heated by the heater 46, to the collection
carrier cartridge 82 to be disposed in the reagent/carrier
container mounting unit 54 to be described in detail below. The hot
water supply nozzle 48 may be configured using a metal (SUS) tube.
Alternatively, the hot water supply nozzle 48 may be configured
using a glass tube or a resin tube. To an end at the side opposite
to an ejection port in the hot water supply nozzle 48, the hot
water supply tube 50 that is connected to peristaltic pump 44
through the heater 46 is connected. A suction-side tube 52 in the
peristaltic pump 44 is connected to the buffer supply unit 64 to be
described in detail below.
[0049] According to the hot water supply unit 42 having the above
configuration, the hot water can be continuously ejected from the
hot water supply nozzle 48, by driving the peristaltic pump 44.
[0050] The reagent/carrier container mounting unit 54 is a stage to
dispose the reagent or the collection carrier used in the
luminescence measurement. In the reagent/carrier container mounting
unit 54, a collection carrier cartridge holder 56, a reagent rack
58, a luminescence measuring tube holder 60a, and a water discharge
port 100 are disposed. The collection carrier cartridge holder 56
is a holder to set the collection carrier cartridge 82. The
collection carrier cartridge holder 56 is provided with a heat
block including a heater and heats the set collection carrier
cartridge 82.
[0051] In the reagent rack 58, a reagent cartridge where the
reagent used in the luminescence measurement is filled is disposed.
As shown in FIGS. 5A and 5B, the reagent cartridge is a package
where reagents of different kinds and pure water are filled into
concave parts (9 parts in an example shown in FIG. 5B) partitioned
in plural parts. An upper opening of each concave part is sealed by
an aluminum sheet (film). By this configuration, the reagent is not
exposed to the outside, until the aluminum sheet is removed, and
the reagent that is placed in stock is not contaminated by viable
bacteria. FIG. 5A is a top view of the reagent/carrier container
mounting unit 54 and FIG. 5B is a top view of the reagent cartridge
62.
[0052] In the luminescence measuring tube holder 60a, a
luminescence measuring tube 60 is disposed. The luminescence
measuring tube 60 is a micro tube that executes a luminescence
reaction of the ATP extracted from the viable bacteria collected by
the collection carrier cartridge 82.
[0053] The water discharge port 100 is a disposable port to discard
the pure water corresponding to the operation fluid of the reagent
dispensing nozzle 24 or the pure water from the hot water supply
nozzle 48. The water discharge port 100 has an operation fluid
discharge position 102 to discharge the operation fluid from the
reagent dispensing nozzle 24 and a hot water discharge position 104
to discharge the hot water from the hot water supply nozzle 48. By
regularly or periodically discharging the pure water collected in
the nozzle, generation of the bacteria in the nozzle and
contamination of the nozzle can be prevented. Further, an increase
of the contamination in the device or generation of cross
contamination can be prevented.
[0054] The buffer supply unit 64 has a reagent dispensing nozzle
control water tank (hereinafter, simply referred to as control
water tank 66) and a hot water supply water tank 68. In a process
after the reagent dispensing nozzle 24 is used, since a process of
removing isolated ATP is not included, the water (pure water) in
the control water tank 66 that is filled into the dispensing
operation tube 30 to link the syringe bump 32 and the reagent
dispensing nozzle 24 needs to have cleanness higher than the water
(pure water) in the hot water supply water tank 68. For this
reason, the control water tank 66 is configured to have a small
capacity and appropriately exchange the stored water, as compared
with the hot water supply tank 68. Since the water in the hot water
supply water tank 68 flows into the collection carrier cartridge 82
set to the collection carrier cartridge holder 56, the hot water
supply water tank 68 needs to have the large capacity, as compared
with the collection carrier cartridge holder 56.
[0055] The control water tank 66 that is set in the above-described
way is connected to the distribution valve 40 in the dispensing
operation tube 30 by the buffer supply tube 70, and the pure water
can be supplied to the dispensing operation tube 30 by switching
the distribution valve 40. The hot water supply water tank 68 is
connected to the suction side of the peristaltic pump 44 and is
sucked by driving the peristaltic pump 44.
[0056] The filter unit 72 removes the collection carrier in the
collection carrier cartridge 82 that is diluted by the hot water
ejected from the hot water supply nozzle 74. The filter unit 72 is
configured using a suction pump 74 and a suction head 76 as a basic
body. The suction pump 74 is a pump that generates the negative
pressure in the suction head 76 to be described in detail below.
The suction head 76 is a tubular body where a front end is
opened.
[0057] In the filter unit 72 that has the above configuration, a
tip end is connected to a lower part of the collection carrier
cartridge holder 56, and the collection carrier that is diluted by
the hot water can be sucked and removed through a collection filter
90 (refer to FIG. 6) by operating the suction pump 74.
[0058] A PMT unit 78 measures the luminescence amount of the ATP in
the luminescence measuring tube 60. In the measuring unit 12
according to this embodiment, the PMT unit 78 is configured in a
head-on type and is disposed on the lower part of the luminescence
measuring tube 60. By this configuration, the light that is
generated in the luminescence measuring tube 60 is incident from an
upper part of the PMT unit 78 and the luminescence amount is
measured.
[0059] The control unit 11 is an element that controls the
components with respect to the input value to the luminescence
measuring device and automates the luminescence measurement.
[0060] The collecting unit 80 is a device that collects the viable
bacteria in the air in the collection carrier cartridge 82. The
collecting unit 80 is configured using a collection carrier
cartridge 82, a blast fan 84, an impactor nozzle head 86, and a
discharge filter 88 as a basic body.
[0061] The collection carrier cartridge 82 collects the viable
bacteria that float in the air. The collection carrier cartridge 82
includes a collection carrier 82a (refer to FIG. 6) to collect the
viable bacteria. The collection carrier 82 that is included in the
collection carrier cartridge 82 according to this embodiment forms
a gel shape at the normal temperature and is solated by heating. In
a lower part of the collection carrier 82a, a cavity (not shown) to
fill diluting hot water is provided. A lower part of the cavity
includes a collection filter 90 (refer to FIG. 6) that filters the
hot water diluting the collection carrier 82a.
[0062] The blast fan 84 sucks air in the collecting unit 80 and
collides the collecting carrier 82a in the collection carrier
cartridge 82 with the floating bacteria in the air. The blast fan
84 is preferably disposed on the downstream side (lower part side
to use an upper part as a suction port in the collecting unit 80
according to this embodiment) of the arrangement position of the
collection carrier cartridge 82. In the collecting unit 80, the
amount of air to be collected can be determined from the blast
amount of the blast fan 84 and the operation time.
[0063] The impactor nozzle head 86 is disposed on an upper part of
the collecting unit 80 and functions as a cover and an accelerator
of the collection carrier cartridge 82. In order to collide the
collection carrier cartridge 82 with the viable bacteria, the flow
velocity of the air that flows into the collecting unit 80 needs to
be fast to some degree. However, the blast fan 84 needs to be
formed to have a large size or have a high rotation speed to obtain
the high flow velocity, and a size of the collecting unit may be
increased.
[0064] In the impactor nozzle head 86, plural ports with the small
diameter are provided, and the air that is sucked by the blast fan
84 passes through the ports with the small diameter and collide the
collection carrier 82a. When the flow volume of the air is
constant, the flow velocity of the passed fluid can be increased by
narrowing an area of a flow passage. For this reason, the needed
flow velocity can be obtained without increasing the size or the
rotation speed of the blast fan 84.
[0065] The discharge filter 88 is disposed on the downstream side
(lower side in the collecting unit 80 in this embodiment) of the
blast fan 84 and removes dust that is contained in the discharged
air.
[0066] By this configuration, the collecting unit 80 according to
this embodiment can have a small size and light weight.
[0067] In the luminescence measuring device 10 with the above
configuration that includes the measuring unit 12 and the
collecting unit 80, first, the viable bacteria in the air are
collected by the collecting unit 80 (step 100: refer to FIG.
6).
[0068] Next, the collection carrier cartridge 82 where the viable
bacteria are collected is extracted from the collecting unit 80 and
the collecting unit 80 is set to the collection carrier cartridge
holder 56 of the measuring unit 12. The collection carrier
cartridge 82 that is set to the collection carrier cartridge holder
56 is heated by the heat block. By the heating, the collection
carrier is solated. The solated collection carrier 82a is sucked
and removed by the filter unit 72 through the collection filter 90,
and the viable bacteria and the free ATP that are collected in the
collection carrier 82a remain in the collection filter (step 110:
refer to FIG. 6).
[0069] After the collection carrier 82a is filtered, the free ATP
is removed and a viable bacteria sample is split by operating the
reagent dispensing unit 14. First, the (ATP removal) reagent is
split from the reagent cartridge 62 by the reagent dispensing
nozzle 24, the reagent is dispensed to the collection carrier
cartridge 82, and the free ATP is removed. By this work, the
measurement error of the luminescence amount can be prevented from
being generated due to the luminescence reaction caused by the free
ATP. Next, the (ATP extraction) reagent is dispensed on the
collection filter 90 in the collection carrier cartridge 82 after
the free ATP is removed, and the ATP is extracted from the viable
bacteria on the collection filter 90 (step 120: refer to FIG.
6).
[0070] The ATP extraction sample is split from the collection
filter 90 in the collection carrier cartridge 82 and is dispensed
to the luminescence measuring tube 60. In the luminescence
measuring tube 60, the luminescence reagent is dispensed in
advance, and the luminescence reaction starts at the same time as
the dispensing of the ATP extraction sample. In the luminescence
reaction in the luminescence measuring tube 60, the luminescence
strength is measured by the PMP unit 78 (step 130: refer to FIG.
6).
[0071] In the luminescence measuring device 10 that has the above
configuration, the process from the splitting of the viable
bacteria sample from the collection carrier cartridge 82 to the
measurement of the luminescence amount is automatically executed in
the measuring unit 12 that is covered with the outer shell. For
this reason, the viable bacteria sample is rarely affected by the
contamination. After the luminescence reagent is previously
dispensed to the luminescence measuring tube 60 set to the
reagent/carrier container mounting unit 54, the ATP extraction
sample from the viable bacteria is dispensed. For this reason, self
background light of the reagent can be measured. For this reason, a
relationship between the luminescence amount and the luminescence
time can be accurately obtained, and calculation of the ATP amount
based on the luminescence amount, that is, measurement of the
number of viable bacteria can be performed with high precision.
[0072] Next, the reagent splitting/dispensing mechanism according
to this embodiment will be described with reference to FIGS. 7 and
8. FIGS. 7A to 7G are state views showing an aspect of reagent
splitting/dispensing based on the reagent dispensing nozzle in the
measuring unit. FIG. 8 is a flowchart showing each operation of the
reagent splitting/dispensing mechanism. The reagent
splitting/dispensing mechanism according to this embodiment is
configured to include the reagent dispensing unit 14 and the
control unit 11. In the reagent splitting/dispensing mechanism that
has the above configuration, the reagent is split/dispensed as
follows.
[0073] The reagent splitting/dispensing operation based on the
reagent dispensing nozzle 24 is executed on the basis of a drive
signal from the control unit 11 with respect to various actuators.
First, in a state where the reagent dispensing nozzle 24 is at the
waiting position, the control unit 11 outputs a drive signal (first
air layer arrangement signal) to pull out the plunger 36 and
generate the negative pressure in the syringe 34 to the actuator 38
of the syringe pump 32. The actuator 38 receives the first air
layer arrangement signal and is driven. As a result, the pure water
that is the operation fluid filled into the dispensing operation
tube 30 and the reagent dispensing nozzle 24 flows into the syringe
34 of which the pressure becomes the negative pressure. Thereby, an
occupied area of the operation fluid in the reagent dispensing
nozzle 24 is decreased, and a first air layer 110 is disposed
between an interface of the operation fluid and a nozzle tip end in
the reagent dispensing nozzle 24. By disposing the first air layer
110 in the nozzle tip end, the operation fluid is prevented from
vertically falling from the nozzle tip end in the waiting state.
For this reason, generation of the cross contamination due to
falling of a droplet or deterioration of splitting/dispensing
precision due to the change in the reagent concentration can be
prevented.
[0074] The first air layer 110 that is disposed in the waiting
process preferably secures the sufficient capacity with respect to
the volume of the reagent dispensing nozzle 24. In this case, the
sufficient capacity may be a ratio of about 30 to 120% of the
nozzle capacity. When the nozzle capacity is set as 20 pi, the
capacity of the first air layer 110 may be set as 100 pi (refer to
FIG. 7A: step 200: waiting process).
[0075] Next, the control unit 11 outputs a drive signal (first
drive signal) to move the reagent dispensing nozzle 24 to the
position right above the concave part where the reagent becoming
the splitting object in the reagent cartridge 62 is filled, to the
X-axis mechanism unit 20 and the Y-axis mechanism unit 18 in the
triaxial actuator 16. In the waiting process described above, since
the air layer (first air layer 110) having the sufficient capacity
with respect to the volume of the reagent dispensing nozzle 24 is
disposed in the tip end of the reagent dispensing nozzle 24, the
operation fluid does not fall at the time of moving as well as
waiting (step 210: first moving process).
[0076] Next, the control unit 11 outputs a drive signal (air layer
adjustment signal) to push the plunger 36 and to generate the
positive pressure in the syringe 34 to the actuator 38 of the
syringe 32. The actuator 38 receives the air layer adjustment
signal and is driven. As a result, the operation fluid is
discharged from the syringe 34 of which the pressure becomes the
positive pressure, and the operation fluid flows into the reagent
dispensing nozzle 24 through the dispensing operation tube 30.
Thereby, an occupied area of the operation fluid in the reagent
dispensing nozzle 24 is increased. In this case, the air layer
adjustment signal drives the actuator 38, such that a part of the
first air layer 110 is discharged from the reagent dispensing
nozzle 24 and the other part remains in the tip end of the reagent
dispensing nozzle 24. That is, the air layer adjustment signal
causes the movement amount of the plunger 36 based on the air layer
adjustment signal to be smaller, as compared with the first air
layer arrangement signal (refer to FIG. 7B: step 220: air layer
adjusting process).
[0077] Next, the control unit 11 outputs a drive signal (second
movement signal) to descend the reagent dispensing nozzle 24 and
deposit the nozzle tip end in the reagent to the Z-axis mechanism
unit 22 in the triaxial actuator 16 (step 230: second moving
process).
[0078] Next, the control unit 11 outputs a drive signal (reagent
split signal) to pull out the plunger 36 and generate the negative
pressure in the syringe 34 to the actuator 38 of the syringe pump
32. The actuator 38 receives the reagent split signal and is
driven. As a result, the operation fluid that is filled into the
dispensing operation tube 30 and the reagent dispensing nozzle 24
flows into the syringe 34 of which the pressure becomes the
negative pressure. Thereby, an occupied area of the operation fluid
in the reagent dispensing nozzle 24 is decreased and the reagent
that becomes the splitting object flows from the tip end of the
reagent dispensing nozzle 24. By this operation, the first air
layer 110 is interposed between the operation fluid and the
reagent. Thereby, the operation fluid is not mixed with the reagent
and generation of the cross contamination due to mixing and
diffusing of the reagent or deterioration of dispensation precision
of the reagent can be prevented (refer to FIG. 7C: step 240:
reagent splitting process).
[0079] Next, the control unit 11 outputs a drive signal (third
movement signal) to ascend the reagent dispensing nozzle 24 and
evacuate the nozzle tip end from the reagent to the Z-axis
mechanism unit 22 in the triaxial actuator (step 250: third moving
process).
[0080] Next, the control unit 11 outputs a drive signal (reagent
protection signal) to pull out the plunger 36 and output generate
the negative pressure in the syringe 34 to the actuator 38 of the
syringe pump 32. The actuator 38 receives the reagent protection
signal and is driven. As a result, the operation fluid that is
filled into the dispensing operation tube 30 and the reagent
dispensing nozzle 24 flows into the syringe 34 of which the
pressure becomes the negative pressure. Thereby, an occupied area
of the operation fluid in the reagent dispensing nozzle 24 is
decreased and the second air layer 112 is disposed between the
interface of the reagent disposed in the tip end of the reagent
dispensing nozzle 24 and the nozzle tip end. Thereby, dispensation
precision can be prevented from being deteriorated due to vertical
falling of a droplet of the reagent with respect to the nozzle tip
end and the cross contamination can be prevented from being
generated due to falling of the droplet (refer to FIG. 7D: step
260: reagent protecting process).
[0081] Next, the control unit 11 outputs a drive signal (fourth
movement signal) to move the reagent dispensing nozzle 24 to the
reagent dispensation position to the X-axis mechanism unit 20 and
the Y-axis mechanism unit 18 in the triaxial actuator 16. In this
case, the reagent dispensation position means an upper part of the
collection carrier cartridge holder 56 or an upper part of the
luminescence measuring tube 60 (step 270: fourth moving
process).
[0082] Next, the control unit 11 outputs a drive signal (reagent
dispensation signal) to push the plunger 36 into the actuator 38 of
the syringe pump 32 and to cause the positive pressure to be
generated in the syringe 34. The actuator 38 receives the reagent
dispensation signal and is driven. As a result, the operation fluid
is discharged from the syringe 34 of which the pressure becomes the
positive pressure, and the operation fluid flows into the reagent
dispensing nozzle 24 through the dispensing operation tube 30.
Thereby, an occupied area of the operation fluid in the reagent
dispensing nozzle 24 is increased. In this case, the reagent
dispensation signal drives the actuator 38, such that the reagent
filled into the reagent dispensing nozzle is ejected and a part of
the first air layer 110 remains in the tip end of the reagent
dispensing nozzle. By this operation, the split reagent can be
completely dispensed and the operation fluid can be prevented from
being ejected (refer to FIG. 7E: step 280: reagent dispensing
process).
[0083] Next, the control unit 11 outputs a drive signal (temporary
evacuation signal) to move the reagent dispensing nozzle 24 to the
operation fluid discharge position 102 in the water discharge port
100 to the X-axis mechanism unit 20 and the Y-axis mechanism unit
18 in the triaxial actuator 16 (step 290: temporary evacuating
process).
[0084] Next, the control unit 11 outputs a drive signal (operation
fluid ejection signal) to push the plunger 36 into the actuator 38
of the syringe 32 and to cause the positive pressure to be
generated in the syringe 34. The actuator 38 receives the operation
fluid ejection signal and is driven. As a result, the operation
fluid is discharged from the syringe 34 of which the pressure
becomes the positive pressure, and the operation fluid flows into
the reagent dispensing nozzle 24 through the dispensing operation
tube 30. The amount of flowing operation fluid exceeds the allowed
amount of the nozzle and a part of the operation fluid is ejected
to the operation fluid discharge position 102. The operation fluid
of the ejected amount is replenished from the control water tank 66
by controlling the distribution valve 40 disposed in the dispensing
operation tube 30 and operating the actuator 38 to pull out the
plunger 36 (refer to FIG. 7F: step 300: operation fluid ejecting
process).
[0085] Next, the control unit 11 outputs a drive signal (operation
fluid protection signal) to pull out the plunger 36 and generate
the negative pressure in the syringe 34 to the actuator 38 of the
syringe pump 32. The actuator 38 receives the operation fluid
protection signal and is driven. As a result, the operation fluid
that is filled into the dispensing operation tube 30 and the
reagent dispensing nozzle 24 flows into the syringe of which the
pressure becomes the negative pressure. Thereby, an occupied area
of the operation fluid in the reagent dispensing nozzle 24 is
decreased and the first air layer 110 is disposed between the
interface of the operation fluid filled into the reagent dispensing
nozzle 24 and the nozzle tip end. In this case, the disposed first
air layer 110 is set to have the same capacity as that of the first
air layer 110 disposed in the waiting process described above
(refer to FIG. 7G: step 310: operation fluid protecting
process).
[0086] Next, the control unit 11 outputs a drive signal (fifth
movement signal) to move the reagent dispensing nozzle 24 to the
waiting position to the X-axis mechanism unit 20 and the Y-axis
mechanism unit 18 in the triaxial actuator 16 (step 320: fifth
moving process).
[0087] When the reagent is dispensed again after the fifth moving
process ends, the repetitive control from the first moving process
(step 210) is executed.
[0088] According to the reagent splitting/dispensing mechanism
where the above control is performed under the above configuration,
when the reagent dispensing nozzle 24 to split/dispense the reagent
is in the waiting state, the operation fluid in the reagent
dispensing nozzle 24 can be suppressed from being contaminated. The
droplet can be prevented from falling at the time of the movement
operation, and the cross contamination can be prevented from being
generated. Thereby, the reagent splitting/dispensing operation can
be performed with high precision.
* * * * *