U.S. patent application number 13/586961 was filed with the patent office on 2012-12-13 for micro-calorimeter apparatus.
This patent application is currently assigned to SENZIME AB (PUBL.). Invention is credited to Thomas CARLSSON, Kjell ROSENGREN.
Application Number | 20120315691 13/586961 |
Document ID | / |
Family ID | 22821316 |
Filed Date | 2012-12-13 |
United States Patent
Application |
20120315691 |
Kind Code |
A1 |
CARLSSON; Thomas ; et
al. |
December 13, 2012 |
MICRO-CALORIMETER APPARATUS
Abstract
A micro-calorimeter apparatus includes a thermostated housing
(3,4,5); a pair of essentially flat heat sinks (9,10), suspended in
the housing (2) and thermally floating relative to the environment
inside the housing (3,4,5). The heat sinks (9,10) are arranged with
their surfaces facing each other. A pair of Peltier elements (11)
are thermally attached to the heat sinks (9,10), one element (11)
on each heat sink (9,10), on the facing surfaces, forming a gap
between them for the accommodation of a generally flat biosensor
unit (12).
Inventors: |
CARLSSON; Thomas; (UPPSALA,
SE) ; ROSENGREN; Kjell; (HASSELBY, SE) |
Assignee: |
SENZIME AB (PUBL.)
UPPSALA
SE
|
Family ID: |
22821316 |
Appl. No.: |
13/586961 |
Filed: |
August 16, 2012 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10333410 |
Aug 20, 2003 |
8262989 |
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PCT/SE01/01664 |
Jul 23, 2001 |
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13586961 |
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60219932 |
Jul 21, 2000 |
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Current U.S.
Class: |
435/287.1 ;
422/51 |
Current CPC
Class: |
G01N 25/4873 20130101;
G01K 17/006 20130101; G01N 25/4826 20130101 |
Class at
Publication: |
435/287.1 ;
422/51 |
International
Class: |
G01N 25/24 20060101
G01N025/24; C12M 1/34 20060101 C12M001/34 |
Claims
1. A generally flat biosensor unit, comprising: a holder portion
(13): a reaction chamber (14), provided as an integral part of the
holder portion (13), and wherein the reaction chamber (14) is an
essentially flat structure with two opposing surfaces between which
the reaction chamber is formed; two openings (15) in the holder for
the inlet and outlet, respectively, of a fluid flow, said openings
being in communication with the reaction chamber via fluid channels
(16).
2. The biosensor unit according to claim 1, wherein the reaction
chamber is covered on each side by a thin foil.
3. The biosensor unit according to claim 2, wherein the thin foil
is made of a polymeric material
4. The biosensor unit according to claim 3, wherein the polymeric
material is a polyimide.
5. The biosensor unit according to claim 2, further comprising at
least one support material between the foils, on which an enzyme is
immobilized, to form an active matrix.
6. The biosensor unit according to claim 5, wherein the thickness
of the active matrix is less than 1 mm.
7. The biosensor unit according to claim 1, wherein the biosensor
unit is made of a polymeric material.
8. The biosensor unit according to claim 1, further comprising an
open structure in the holder in order to minimize thermal
transportation through the holder.
9. The biosensor unit according to claim 1, wherein the holder has
a generally "H"-shaped cross-section, the "H" having legs and a
web, and where the web of the holder that joins the legs of the
"H", has an opening made in the web, and the web is covered on each
side by a thin foil of a polymeric material.
10. The biosensor unit according to claim 9, wherein the polymeric
material is polyimide.
11. The biosensor unit according to claim 1, wherein there is
provided a support material or beads between the foils on which one
or more of the group consisting of an enzyme, antibodies and
antigens can be immobilized, thereby forming an active matrix.
12. The biosensor unit according to claim 1, wherein there are
beads provided between the foils on which one or more of the group
consisting of an enzyme, antibodies and antigens can be
immobilized, thereby forming an active matrix.
13. The biosensor unit according to claim 9, wherein the active
matrix in the reaction chamber is less than 1 mm thick.
14. The biosensor unit according to claim 9, wherein the holder
forms two opposing flat side surfaces oriented essentially
perpendicularly with respect to the flat reaction chamber
structure, and wherein there are through holes provided in the side
surfaces, exiting inside the reaction chamber for the introduction
of a flowing sample.
Description
[0001] The present invention relates to biosensor systems in
general, and in particular to a calorimetric type of system,
especially for enzymatic reactions.
BACKGROUND OF THE INVENTION
[0002] The term biosensor is commonly referred to as a measuring
device that combines a biological sensing part is close proximity
to a non-biological transducer which converts the biochemical or
electrochemical Information produced in the biological part into a
measurable signal.
[0003] The most commonly employed transducer in current biosensor
instruments (e.g. a clinical analyser) today is electrochemically
based. An clinical analyser is often used in connection with a
biosensor unit (one-time use), which before operation, is connected
to the analyser. The biosensor unit contains both the biological
and the electronic transducer. It has turned out that there is a
significant cost of designing and fabricating a custom integrated
biosensor unit that includes both the biological sensing part and
the necessary electronic elements. One major reason for this is
that the materials and methods used for electronic components
require very high temperatures, which temperature levels are
incompatible with the fabrication of the biological part. It has
turned out to be difficult to combine these contradicting demands
of the devices to be manufactured.
SUMMARY OF THE INVENTION
[0004] In order to eliminate the drawbacks associated with the
prior art devices it is suggested according to the present
invention to separate the biological parts from the electronic
parts in the biosensor unit. This can be achieved by using a new
thermal transducer technology.
[0005] When the analysis is to be earned out the biosensor unit
itself is easily connected and brought in thermal contact with the
transducer in the instrument.
[0006] Thereby, only the requirements and limitations set by the
biological system need to be considered in the manufacturing
process of the biosensor unit. Also, packaging of the biosensor
unit should thereby be simplified.
[0007] The invention is defined in claim 1 that defines a
micro-calorimeter apparatus comprising a thermostated housing; a
pair of essentially flat heat sinks, suspended in said housing and
thermally floating relative to the environment inside the housing,
and arranged with their surfaces feeing each other; and a pair of
Peltier elements thermally attached to said heat sinks, one dement
on each heat sink, on said facing surfaces, forming a gap between
them for the accommodation of a generally flat biosensor unit. The
term "generally flat" indicates that at least the reaction chamber
of such a biosensor unit should be flat in the meaning that the
thickness of such a chamber and the opposite large walls enclosing
it is substantially smaller than to the dimensions of said large
walls. The calorimeter as well as the biosensor unit shall be
designed so as to enable a continuous flow of a sample of bio fluid
through the reaction chamber. Therefore, the calorimeter as well as
the biosensor unit comprises suitable channels for this purpose.
Heat produced by the reaction between fluid and enzymes is
transmitted via the opposite large walls of the chamber to the
respective Peltier element, thereby making it possible to measure
the reaction.
[0008] Preferably, the Peltier elements are arranged so as to be in
a heat conducting contact with thin foils defining opposite walls
of a reaction chamber defined by the biosensor unit when the
biosensor unit is in an operative position between the Peltier
elements. Inside the reaction chamber, enzymes adapted for the
reaction are arranged. A basic principle of the invention is that
the heat transmitting area between Peltier elements and reaction
chamber walls should be as large as possible in relation to the
volume of the reaction chamber. Therefore, the thickness of the
reaction chamber is small to relation to the dimensions of the
walls defined by the foils.
[0009] For best performance of the calorimeter the heat conducting
contact area between Peltier elements and the foils should
correspond to the total reaction chamber wall area, defined by said
foils. Total chamber wall area is referred to as the total area of
the outer surface of the chamber wall-forming part of the foils
directed towards the respective Peltier element.
[0010] According to a preferred embodiment at least one of the
Peltier elements is movable to and from the other Peltier element
in order to facilitate insertion of the biosensor unit between the
Peltier elements.
[0011] Preferably the housing comprises a plurality of metal
blocks, at least one of which acts as a thermostat element, for the
purpose of providing a generally constant air temperature around
the heat sinks and the Peltier elements during operation of the
calorimeter.
[0012] The heat sinks are suspended in said housing by means of
heat insulating members connected to a respective block.
[0013] At least one of the heat sinks should be moveable in
relation to the block connected thereto along the insulating member
on which it is suspended.
[0014] Preferably, the calorimeter comprises a force element, such
as a spring, exerting a force, preferably a spring force, on the
movable heat sink towards the other heat sink for the purpose of
pressing an inserted biosensor unit against the other heat sink and
belonging Peltier element. The force element spring also enables
withdrawal of the movable heat sink/Peltier element for the purpose
of facilitating insertion or removal of the bio sensor unit.
[0015] According to one embodiment, the housing is defined by two
opposite blocks and a cylinder located between said opposing
blocks, each heat sink being connected to a respective one of the
opposing blocks. The cylinder is hollow in order to accomodate the
heat sink/pettier element units as well as the biosensor unit. In
this particular embodiment the insulating members comprise holding
pins made of a material presenting a low heat conductivity.
[0016] Preferably, the inventive calorimeter also comprises members
for snap fitting the biosensor unit in its operative position,
thereby facilitating fitting of the biosensor unit in the
calorimeter.
[0017] The invention also relates to a biosensor unit,
characterized in that it comprises a generally flat reaction
chamber the external surfaces of which are adapted to fit snugly
between the Peltier elements of a calorimeter apparatus as defined
above.
[0018] Further features and advantages of the invention are
presented in fee following detailed description of preferred
embodiments and in fee annexed patent claims.
BRIEF DESCRIPTION OF THE DRAWINGS
[0019] FIG. 1 is an exploded view, showing a biosensor apparatus
according to the present invention;
[0020] FIG. 3 illustrates the biosensor apparatus with an
insulating house attached;
[0021] FIG. 3a shows a holder portion of the biosensor apparatus
with a reaction chamber open;
[0022] FIG. 3b illustrates the holder portion with the reaction
chamber covered with a thin foil, seen from opposite direction in
relation to FIG. 3a;
[0023] FIG. 4a is an axial cross sectional view of the biosensor
apparatus;
[0024] FIG. 4b is a close-up view of a biosensor unit according to
the present invention In thermal contact with Peltier elements;
[0025] FIG. 4c is a cross sectional view of the biosensor unit
positioned to a pin 21;
[0026] FIG. 5 is a perspective view, partly broken away, showing an
alternative embodiment of the biosensor apparatus according to the
invention;
[0027] FIG. 6a is a perspective, view of the alternative biosensor
unit;
[0028] FIG. 6b is a schematic cross-section through an alternative
embodiment of the reaction chamber according to the present
invention;
[0029] FIG. 7 is a schematic illustration of the positioning of the
alternative biosensor unit.
DETAILED DESCRIPTION OF THE INVENTION
[0030] The principle of calorimetry, i.e. the detection of heat of
reaction, on which the present invention is based is well known
since many years.
[0031] The apparatus according to the invention is schematically
illustrated in FIG. 1 and is generally described with 1. The
apparatus comprises a thermostated space or compartment 2, formed
from, or defined by, two opposing blocks 3,4 and a cylinder 5,
where the block 3 is axially moveable a short distance in relation
to the cylinder 5 and-the block 4 is fixed to the cylinder 5. The
two blocks and the cylinder is separated from each other by
separating members, here small washers 6, thereby forming an
insulating air gap between them. Each part is individually
temperature controlled. The entire device is enclosed in an
insulating house 7, see FIG. 2. The two blocks are provided with,
holding pins 8a,8b made of a material with low heat conductivity,
preferably plastics onto which are placed two heat sinks 9,10, heat
sink 9 being slightly moveable in relation to its block 3 along the
two holding pins 8b, whereas the heat sink 10 is fixed to its
block.
[0032] The heat sinks are circular plates made of a material having
a large heat capacity, such as aluminium. The heat sinks are
located so as to form a space between them. They are also provided
in such a way that they "thermally float" with respect to the
compartment, by fee provision of an air gap surrounding the heat
sinks. The expression "thermally floating" should be taken to mean
that the heat exchange between the heat sink and the thermostated
compartment are kept minimal. This is achieved by suspending the
heat sinks inside the compartment onto the holding pins. On each
heat sink there is mounted a Peltier element 11 In very good
thermal contact with the heat sink. The Peltier elements are
attached to the heat sinks such that they face each other. The unit
of heat sink/Peltier element attached to one 3 of the blocks 3,4 is
to some extent, also movable.
[0033] The biosensor unit 12 shown in FIG. 3a to be used with the
apparatus comprises a holder portion 13 and a reaction chamber 14,
provided as an Integral part of the holder portion 13, The reaction
chamber 14 is an essentially flat structure with two opposing
surfaces between which the reaction chamber is formed. Inlet and
outlet of fluid flow is achieved through two openings 15 in the
holder and the fluid channels 16, etched in the holder. In order to
minimize thermal transportation through the holder it has an open
structure 17, in it.
[0034] Here is shown one example of a biosensor unit design. The
overall detailed design of the biosensor unit with holder, reaction
chamber and fluid channels is a matter of which type of specific
reaction it is applied for. The example shown, here is
approximately 50.times.10.times.0.5 mm, but fee right dimensions
and overall design should be optimised, for each special case.
[0035] The reaction chamber 14 is covered on each side by a thin
foil 20 (see FIG. 3b) made of a polymeric material, e.g. a
polyimide. This polymer is very strong and can thus be made very
thin, yielding a very high rate of heat conduction through it,
which is essential, for the invention. Between the foils 20 there
can be provided support materials an which e.g. an enzyme can be
immobilized, by methods known in the art. The distance between the
surfaces should be as small as possible, and the lower limit is
only set by manufacturing/technological limitations. A practical
upper limit for the thickness of the active matrix in the reaction
chamber is probably about 1 mm.
[0036] The biosensor unit is preferably made of a polymeric
material. Any material that can be easily processed to the desired
structure is suitable.
[0037] The thickness of the reaction chamber 14 (from one external
surface to another) corresponds closely to the distance between the
opposing Peltier elements, for reasons to be discussed below.
[0038] The apparatus according to the invention is operated as
follows.
[0039] By virtue of the heat sink/Peltier element aggregate being
attached to the moveable block 3, it can easily be moved by the
decompression of springs 18, such that the gap between the
aggregates is slightly widened (see FIG. 4a). Thereby it will be
an. easy matter to insert the biosensor unit 12 between them. When
the holder has been inserted to an extent that grooves 19 of the
holder mid a pin 21 meet, the biosensor unit will be "snap-fitted"
in position. Grooves 19 and pin 21 thereby define snap fitting
members. When the biosensor unit is in its block 3, it is moved
together with its heat sink/Peltier element by compression of the
three springs 18. When the block is In Its inner position its heat
sink/Peltier element press, by action of the springs 18, the
biosensor unit to the fixed heat sink/Peltier element. Then the two
Peltier elements will rest against the external surfaces of the
reaction chamber in very good thermal contact. The reaction chamber
of the biosensor unit and the Peltier elements will only be
separated by the thin foils 20 mentioned above, thereby ensuring
excellent heat transfer from the reaction chamber to the Peltier
elements.
[0040] The alternative biosensor apparatus 30 according to the
invention is schematically shown in FIG. 5 (the upper portion
omitted for clarity). It comprises a thermostated space or
compartment 40 formed from a number of metal blocks 60, 80, 100, at
least one of which, preferably two, act as thermostat elements 80,
100, The entire device is enclosed in an insulating housing (not
shown). Two opposing blocks 60 (only one shown) have depressions
120 formed in the surfaces thereof in which there are provided two
movable heat sinks 140 (only one shown). The heat sinks are
circular plates made of a material having a large heat capacity,
such as aluminium.
[0041] The heat sinks are located so as to form a space between
them. They are also provided in such a way that they "thermally
float" with respect to the compartment, by the provision of an air
gap surrounding the heat sinks. This is achieved by suspending the
heat sinks inside the compartment in insulating yokes 160. The
yokes are journalled on shafts 180 extending from the thermostat
blocks 80, 100, On each heat sink there is mounted a Peltier
element 200 in very good thermal contact with the heat sink. The
Peltier elements are attached to the heat sinks 140 such that they
face each other. Each unit of heat sink/Peltier element is to some
extent movable, by the yokes 160 being pivotally mounted as
indicated above,
[0042] The journaling shafts 180 have a further function, namely to
provide flow channels 220 (indicated with broken lines) for a
flowing medium to be analysed in the instrument. The flow channels
extend through the thermostat blocks and terminates in a
spherically shaped end surface 240 located at the end of said
shafts.
[0043] The alternative biosensor unit 260 according to the
invention is shown in FIG. 6a to be used with the alternative
biosensor apparatus 30 and comprises a holder portion 280 and a
reaction chamber 300 provided as an integral part of the holder.
The reaction chamber is an essentially flat structure with two
opposing surfaces between which the reaction compartment is formed.
The holder has a generally "H"-shaped cross-section where the web
320 joining the legs of the "H" has an opening made in it. The web
portion is covered on each side by a thin foil 340 (shown in FIG.
6b) of a polymeric, preferably non-electrically conducting
material, e.g, polyimide. This polymer is strong and can thus be
made thin (not more than half, preferably not more than 1/10 of the
thickness of the reaction chamber (here 0.3-0.5 mm)), thereby
having a high rate of heat conduction through it, which is
essential for the invention. Between the foils there can be
provided a support material, for example beads 360, on which e.g.
an enzyme, anti bodies or anti genes can be immobilized, by methods
known in the art. This general principle is also preferably applied
to the first embodiment described above. The distance between the
surfaces should be as small as possible, and the lower limit is
only set by manufacturing/technological limitations. A practical
upper limit for the thickness of the active matrix in the reaction
chamber is probably about 1 mm. The sensor structure is preferably
made of a polymeric material. Any material feat can be easily
processed to the desired structure is suitable.
[0044] The holder forms two opposing flat side surfaces 320
oriented essentially perpendicularly with respect to the flat
reaction chamber part, in these side surfaces there are through
holes 380 provided, exiting Inside the reaction chamber 300 for the
introduction of a flowing sample. The channel openings on the side
surfaces of the holder are slightly recessed 330 to match, the
spherically shaped end part 240 of the shaft. The thickness of the
reactor (from one external surface to another) corresponds closely
to the distance between the opposing Peltier elements, for reasons
to be discussed below.
[0045] The apparatus according to the invention is operated as
follows.
[0046] By virtue of the heat sink/Peltier element aggregates being
pivotally hinged, they can easily be moved such that the gap
between them is slightly widened (see FIG. 7). Thereby it will be
an easy matter to insert the biosensor structure 260 between there.
When the holder has been inserted to an extent that the recesses
330 of the channel openings and the protrusions 240 meet, the
sensor will be "snap fitted" in position, and then the Peltier
elements 200 will rest against the external surfaces of the reactor
chamber in very good thermal contact.
[0047] The heat developed in the reaction chamber by e.g. an enzyme
reaction will give rise to a heat Sow from the reaction chamber
(hot side) to the heat sinks (cold side). This heat flow will be
taken up and dissipated very quickly by the heat sinks, and
eventually released from them very slowly. This is made possible by
selecting the heat capacity of the heat sinks such that the thermal
impact from, the heat flow on the heat sinks is minimized. In the
present arrangement the thermal disturbance is of the order of
<1-100 ppm. Due to the large heat capacity of the heat sinks the
heat evolved in the reaction chamber will in practice not affect
the temperature in the reaction chamber. Because the heat sinks are
"thermally insulated" from, their immediate environment they will
provide for a uniform heat flow being created from the reaction
chamber to the heat sinks, in connection with the development of
heat in the reaction chamber.
[0048] The heat flow gives rise to a temperature gradient across
the Peltier elements. By operating the Peltier elements in reverse
mode the flow of heat will generate a current from the Peltier
elements which is proportional to the heat development in the
reaction chamber.
[0049] Since the flow of heat horn a body is proportional to its
area it is advantageous to design the reaction chamber and the
Peltier elements in such a way that a maximum contact area is
created between them. In the first embodiment conventional (flat)
Peltier elements have been employed, but in principle any geometry
could be used.
[0050] In use of the biosensor unit a sample flow is fed through
it, and unavoidably some heat from the reaction will escape from
the reaction chamber with the outflow. However, by the suggested
flat design of the reaction chamber substantially all heat from the
reaction will be absorbed by the Peltier elements, and thus only a
minute portion will leak out. Thereby a very high efficiency is
obtained.
[0051] The geometry of the biosensor unit with regard to the
analyte should be optimised for specific situations. As indicated
above the biosensor unit can be designed in many ways, but from a
production point of view, a flat configuration is probably
preferable.
[0052] By virtue of the thermal separation of the heat sinks from
the thermostated environment, the influence of external
disturbances will also be reduced. This together with the fact that
Peltier elements have a very low impedance, enables a very good
signal to noise (S/N) ratio to be achieved. In addition, Peltier
element do not require an external power source for excitation,
which is the case for e.g. electrochemical biosensors.
[0053] Prior art thermal biosensors have utilized thermistors or
thermo-couples as the temperature sensitive elements (transducer).
Both of these have the drawback of a high impedance, and therefore
susceptible of picking up noise.
[0054] By employing the design principles disclosed above, it is
possible to construct a biosensor unit based on thermal detection
with simplified production and with increased sensitivity, thereby
simplifying the production process.
[0055] As an example a sample volume of e.g. 1 .mu.l glucose with
the concentration of 2 mmol/l (a low value in this regard) will
yield a S/N in one of the present biosensor units of approximately
25:1. The corresponding value for thermistor based biosensors is
rarely better than 5:1.
[0056] The simplified manufacturing process and the improved
sensitivity are the main advantages of the present invention.
[0057] It should be emphasized that obvious combinations of
features of the embodiments described above are within the scope of
the invention. The scope of the invention is defined by the
appended claims supported by the description and the annexed
drawings.
* * * * *