DE102006030380A1 - Cool stamp arrangement for adjusting temperature profile in process area for reaction chamber of assay-processor-cartridge for biological sample analysis, comprises cool stamp, isolated metal body and Peltier element arranged on the stamp - Google Patents

Abstract

The cool stamp arrangement for adjusting temperature profile in a process area for reaction chamber of an assay-processor-cartridge for the analysis of biological samples, comprises a cool stamp, an isolated metal body that is movably arranged linear to the surface of the process area to be cooled, a cool stamp area for the reception of a linear drive for viewing a controllable forward and backward movement, and a Peltier element arranged on the cool stamp with cool body and ventilator. The cool stamp is movable by a center hole and cool area for mounting the Peltier element. The cool stamp arrangement for adjusting temperature profile in a process area for reaction chamber of an assay-processor-cartridge for the analysis of biological samples, comprises a cool stamp, an isolated metal body that is movably arranged linear to the surface of the process area to be cooled, a cool stamp area for the reception of a linear drive for viewing a controllable forward and backward movement, and a Peltier element arranged on the cool stamp with cool body and ventilator. The cool stamp is movable by a center hole and cool area for mounting the Peltier element. A nib shaped secondary valve is movably arranged in the center hole for the cool stamp and is held by a screw. An adapter is connected with the linear drive in such a way that a controllable pressure is exercised over a suitable spring against the cool stamp. A multiphase motor or servo gear motor with spindle- or worm gear, a linear multiphase motor, piezo-linear motor, motor with sprocket and gear rack, solenoid, moving magnet, voice-coil-magnet or motor with cam disk are used as the linear drive. The cooling area of the cool stamp is surrounded with a suitable isolation material. A sensor unit and a heat-conducting element are arranged between the cool stamp and the Peltier element for adjusting temperature. The sensor unit is connected by a heat conductive connection or is clenched or screwed with a suitable connection mechanism. An independent claim is included for procedure for adjusting temperature profile in a process area for reaction chamber of an assay-processor-cartridge.

Description

The Invention refers to a cooling stamp device and a method of controlling a temperature profile in one Process chamber, primarily a reaction chamber of an assay processor cartridge (AP cartridge), also referred to as biochar.

Of the Biochip, a fingernail-sized one Microlaboratory with numerous tiny reaction fields is an important one Future tool for the analysis of biological samples.

On a biochip lets within the shortest possible time Time a sample for the presence of a variety to be detected Check molecules.

To the Processing, the biochip is enclosed in the sample space of a cartridge, where he heated according to the investigations in a defined manner can be. The coupling and removal of heat in the reaction space can be done in different ways, such as by internal or external resistance heating, internal induction loops or Areas, by water cooling and heating, by irradiation with light, in particular IR light, via external Microwave radiation, by air cooling and / or heating, by friction, introduced by thermal radiators and by Peltier elements.

To the faster cooling a heated process space, such as the reaction chamber an AP cartridge, various cooling devices are known.

Thus, the cooling of the reaction chamber via air flow (room air or compressed air), blown directly onto a wall of the reaction vessel, generated for example with 12V radial fan. Use in the biochip-equipped AP cartridges (Assay Processor) in the POC readers, such as those in the WO 05108604 A2 is described. The cooling of the process unit can preferably also be achieved by permanently tempering the space surrounding the process unit at a reduced temperature and thereby passively cooling the cartridge. Also known are the cooling by a good thermally conductive plate (ceramic or metal) applied to a reaction chamber by blowing with a fan (room air), the walls of the reaction chamber consist only of a thin Kuststofffolie (Cephied, SmartCycler).

Known are further cooling / heating a metal block with Peltier elements, wherein in the metal block a complete microtiter plate with tubes is inserted (individual tubes, Stripes). The conical tubes dive into correspondingly shaped Holes in the metal block for maximum possible Touchpad on. This technology can be found in most thermal cyclers (e.g. Eppendorf, Biometra, Labtech).

Analogous At RotorGene, the metal block with the tubes is air-flowed cooled. Cooling / heating can also by alternating pressing a metal block with tubes or microtiter plates to others different tempered metal blocks respectively. The metal blocks surrounding the block with the samples become according to the temperatures regulated for the biological processes are necessary.

Farther are in this context the cooling / heating of the sample liquid when flowing through of thin Capillaries, tubes or hoses, which are in different tempered environments (liquids, Air, metal blocks etc.).

disadvantage Some of these known devices are, inter alia, insufficient cooling rates, high technical and financial effort in the realization, high energy input and / or non-reproducible cooling rates.

The cooling down of a body passing by cooler Air is generally very sluggish, as by touch with a cooler solid body or liquid Medium. To similar cool quickly to be able to the air must be blown on the body at high speed and as frontally as possible (e.g., compressed air from nozzles).

The technical implementation of compressed air supply in a portable Device at aspired independence Compressed air home networks is associated with much higher cost than if other cooling principles would be used (e.g., Peltier elements).

This especially applies when temperatures below ambient temperature are sought, the compressed air therefore also be cooled would.

• – muss ein dauerhafter Kontakt des Probenraumes mit einem beheizbaren/kühlbaren Körper/Medium während des gesamten Prozesses hergestellt werden, um den zu temperierenden Körper abwechselnd exakt auf die unterschiedlichen benötigten Temperaturen zu heizen/kühlen, oder
• – es müssen mehrere exakt auf die verschiedenen Zieltempera turen gebrachte Körper/Medien abwechselnd mit dem Probenraum in Berührung gebracht werden.

When cooling by contact with a cooler (actively cooled) body (or medium)

• - A permanent contact of the sample chamber with a heatable / coolable body / medium must be made during the entire process in order to heat / cool the body to be tempered alternately exactly to the different temperatures required, or
• - It must be brought exactly to the different target tempera tures body / media alternately with the sample space in contact to be brought.

at The first variant is needed because of the thermal inertia of the heatable / coolable body / medium and the permanent Change between heating and cooling a lot of energy to the desired To achieve dynamics.

At the Heating and cooling with Peltier elements that means, if the desired speed at all because of the extra inertia the Peltiers at Umpolen is reached, an extremely high power consumption.

at Variant 2 is the cooling with a target temperature controlled body at the beginning though quickly, but gets slower, the closer the actual temperature approaches the target temperature, the lower the so Temperature difference of the two bodies is. Until the target temperature So in fact is reached many seconds have passed, which is usually the case only seconds holding time per temperature step and the many repetitions in some biological processes quickly summed up to minutes.

task The present invention is therefore an apparatus and to provide a method whereby on the one hand, the cooling rate significantly improved and a higher Reproducibility and independence of ambient conditions (room air temperature) is achieved, and about that In addition, the controller behavior is optimized, with minimal underrun the target temperatures.

The object is achieved by a cooling stamp device consisting of an insulated metal body ( 12 ) such as copper or aluminum, which is mounted linearly movable relative to the surface to be cooled of the process space, which has a receiving surface for a linear drive and is cooled by a Peltier element with heat sink and fan. 1 shows the cooling stamp device according to the invention.

When Linear drive are, for example stepper motors or servo geared motors with spindle or worm gear, linear stepper motors, piezolinear motors, Motor with pinion and rack, lifting magnet, rotary magnet, voice coil magnet, Motor with cam, etc. suitable. For lifting and rotary magnets If necessary, the high speed of the armature must be damped.

• – a) den eigentlichen Kühlstempel mit zentrischer Bohrung (11) und seitlich wegführender Kühlfläche zum Anbringen des Peltierelementes darstellt und
• – b) einen in der zentrischen Bohrung (13) zum Kühlstempel beweglich angeordneten stiftförmigen Sekundärstößel (14), der von oben durch eine Schraube (15) gehalten wird und mit ihm die gesamte Einheit des Kühlstempels zusammengehalten wird.

In a particular embodiment of the cooling stamp, the cooling stamp consists of two mutually displaceable parts a) and b), wherein

• A) the actual cooling die with centric bore ( 11 ) and laterally dissipating cooling surface for attaching the Peltier element represents and
• - b) one in the central bore ( 13 ) to the cooling punch movably arranged pin-shaped secondary tappets ( 14 ) from above through a screw ( 15 ) is held and with him the entire unit of the cooling stamp is held together.

All cooled surfaces, except the lower stamp surface, are ideally filled with a suitable material (e.g., self-adhesive Foam or cellular rubber mat) insulated to condensation in warm avoid damp environment and save energy.

2 shows a sectional view of the cooling stamp. The rear part of the inner plunger is clamped on the back in a recording of a linear drive and moved by this drive back and forth. In the exemplary embodiment, the cooling plunger is an aluminum cooling plunger of 6 × 6 × 5 mm, as already described with laterally led out cooling surface with Peltier element, heat sink and fan. He has a central hole of 3 mm. The aluminum is not anodised or otherwise treated. The Peltier element ( 18 ) is 25 × 25 mm, standard of the company SuperCool. The finger heat sink ( 19 ), in 5 between the Peltier element ( 18 ) and the fan ( 20 ), standard is 40 × 40 × 12 mm (not shown in the picture). The fan ( 20 ) of 12 V is standard 30 × 30 mm. Furthermore, a Peltier controller TC2812 (Minco, CH) with power supply, a PT100 sensor ( 22 ), Class A, and two square pieces of a double-sided heat-conducting adhesive tape having an area corresponding to that of the Peltier element (standard adhesive tape for PC processors). The design of the cooling stamp has been designed in such a way that it can be used in a functional sample of a POC reader (without cover) with a modified plunger rod ( 3 . 21 ) has been integrated in such a way that no changes need to be made to the reader after removal.

About a small spring ( 17 . 2 and 5 ) presses the upper clamped part (adapter, 16 ) against the cooling plunger. This is done with a linear ball bearing (' 3 . 23 ) on the ram ( 21 ) held. In this way, the cooling pusher with Peltier element, heat sink and fan (not shown) is moved linearly toward and away from the AP cartridge. In 4 is still the plunger lever ( 25 ) recognizable. The PT100 sensor ( 22 ) was applied by means of 2-component epoxy thermal adhesive between the cooling plunger and Peltier on the Wärmeleitblech ( 24 Glued (near the linear bearing or Kühlstößelgalgen). It can also be pressed or screwed or connected in a manner known per se or with a suitable connection mechanism. The temperature of the cooling plunger was controlled by Peltier controller to 15 ° C. 3 shows the positioning of the cooling plunger via a linear ball bearing on the plunger yoke according to the embodiment. 4 shows a POC function pattern with a cooling ram stamp (without Peltier, heat sink, etc.).

At the Stepper motor were after a reference travel (initialization, position 0) programmed two positions: one for the position "cooling", theoretically short "behind" the surface, the chilled should be (so that the cooling plunger with Spring force pressed will) and a position between this and the reference position, in short (3-4 tenths of a millimeter) in front of the surface to be cooled.

It was also the speed for the movements between these two positions increased significantly, as the Standard speed in the functional pattern is otherwise very low. The control of the ram motor had to go for this experiment in the POC function pattern in addition to the control of the heating in the chip of the AP cartridge on the firmware side in the algorithm for the temperature control be involved.

• – Es wurde ein mit einem PT100-Sensor im Probenraum ausgestatteter, mit Wasser gefüllter AP in das Funktionsmuster eingelegt und kontaktiert. Danach wurde der Kühlstempel auf 15°C geregelt und die für den verwendeten Siliziumchip gültigen Kalibrierwerte in der Software des Funktionsmusters eingegeben (damit der AP auf die korrekten Temperaturen geheizt werden kann). In einer in der Fumu-Software lauffähigen Scriptdatei wurde ein Dauertest programmiert (eigene Makrosprache), mit einem Temperaturverlauf, wie er für viele biologische Testsprozeduren typisch ist, zunächst mit 500 Zyklen.
• – Der interne Sensor (Probenraumtemperatur) wurde über einen PT100-Messverstärker (Greisinger Electronic) und ein USB-DAQ-Modul (digitales USB-Datenerfassungsmodul, Meilhaus Electronic) an einen PC angeschlossen und des sen Temperaturverlauf mit einer selbsterstellten Software (LabView^TM – Applikation) in einer Logdatei mitgeschrieben. Der geregelte Temperaturverlauf am Si-Chip und die Solltemperatur wurden von der Funktionsmuster-Software mitgeloggt.
• – Das Script wurde gestartet. Nach 500 erfolgreichen Zyklen wurde die AP-Kartusche neu befüllt, es hatten sich Luftblasen gebildet (die übliche Zyklenzahl eines biologisch relevanten Tests liegt bei 30-40). Die Zykluszahl wurde auf 1000 erhöht und der Dauertest erneut gestartet. Nach weiteren 770 Zyklen mit dem gleichen AP und Si-Chip wurde der Dauertest abgebrochen.

The experimental procedure of the embodiment, whose main objective was to find out how well an aluminum stamp can cool (standard equipment-relevant material), and whether it can short-circuit or damage the sensitive heating and sensor structures applied to the silicon chips of the AP cartridges when pressed, is further described as follows:

• - A water-filled AP equipped with a PT100 sensor in the sample room was placed in the functional pattern and contacted. Thereafter, the cooling stamp was adjusted to 15 ° C and the calibration values valid for the silicon chip used were entered in the software of the function pattern (so that the AP can be heated to the correct temperatures). In a run in the Fumu software script file a duration test was programmed (own macro language), with a temperature history, as it is typical for many biological test procedures, first with 500 cycles.
• - The internal sensor (sample chamber temperature) was connected to a PC via a PT100 measuring amplifier (Greisinger Electronic) and a USB-DAQ module (digital USB data acquisition module, Meilhaus Electronic) and its temperature profile with a self-created software (LabView ^TM application ) in a log file. The controlled temperature profile on the Si chip and the setpoint temperature were logged by the function pattern software.
• - The script has been started. After 500 successful cycles, the AP cartridge was refilled, air bubbles had formed (the usual number of cycles of a biologically relevant test is 30-40). The cycle number has been increased to 1000 and the endurance test restarted. After another 770 cycles with the same AP and Si chip, the endurance test was aborted.

Out 6 you can read the result of the endurance test. All curves are almost congruent, that is, there were no impairment of the heating or sensor structures by pressing the aluminum ram over a total of 1270 cooling and heating processes of a single AP cartridge or silicon chips. 6 shows temperatures on the silicon chip, 3 cycles at the beginning and at the end of the two endurance tests with the aluminum cooling plunger in the functional sample (always the same AP or chip). Figure 7 shows the temperatures in the sample chamber (Pt100 sensor) during the endurance test.

In another laboratory test (not in the functional sample) were similarly good cooling rates achieved with a copper stamp.

The The invention also relates to a method for controlling a temperature profile in a process room. The control of the temperature profile of a process room, such as the reaction chamber of an AP cartridge by the method according to the invention according to the features of claim 10 and the dependent subclaims. Of the Process or sample space of an AP cartridge is indicated by a Heated heating with sensor element heated. The control of the faster and targeted cooling takes place according to the invention by a cooled with Peltier Cooling piston, the significantly colder as the target temperature of the process space is, with a defined Pressure by a linear punch drive to the contact surface of the Process room pressed is and upon reaching the target temperature of the contact surface again is moved away. Advantageously, this is a fast-reacting Drive used in addition defined can be pressed with adjustable pressure. The way of the rest position of the cooling stamp until cooled body or the contact area, can be very small, because the body to be cooled is actively heated and also over a very small air gap hardly transfers heat. It can be a cheaper one Drive (e.g., solenoid).

In an exemplary embodiment, the stamping temperature of the metal body is at 20 ° C to cool at an actual temperature of the process chamber of 90 ° C this to the desired target temperature of 50 ° C. It is important to ensure that advantageously the cooling stamp is not cooled too deep, so that the water from the ambient air can not condense. The measurement of the temperature via a suitable temperature sensor, which is located, for example, with the heating element in the wall to the process room. This also causes in a conventional manner, the control mechanisms for moving the Kühlstempels on the punch drive. The heating in the wall of the process chamber and the drive of the cooling stamp are controlled by the same regulator. The Peltier element and thus the temperature of the cooling plunger is kept constant by a separate controller.

Should cooled after heating be turned off, the heating current and at the same time the punch pressed against the wall (e.g., 10-20 N). This pulls out the heat very quickly the wall and the process room behind it.

In the moment when the temperature of the wall reaches the target temperature the stamp is moved away again and the heating starts again, to correct the temperature. Should, as in some biological tests necessary to set a temperature below room temperature in the reaction space be, the stamp is timely brought to this temperature and then pressed permanently. In special execution is to a limited extent (up to the temperature difference of about 40-50 ° C) against the pressed 15 ° C cold cooling piston heated.

In In another particular preferred embodiment, the linear drive presses in the second position only the cooling plunger over a suitable spring against the chamber wall of a process space (e.g. the Si-chip of the AP cartridge) until the target temperature is reached or falls below.

In a third position just behind the second one is in the central hole ( 13 ) arranged pin-shaped secondary tappets ( 14 ) moved so far in the direction of the process room until it presses with significantly more force than the cooling punch centric and punctiform against the chamber wall. This punctiform applied large pressure force is required, a biochip glued on the other side of the chamber wall in the sample space ( 1 ) to move in the same direction against the force of a seal until this biochip rests against a transparent cover glass on the opposite side of the sample chamber and a sharp image of it can be taken with a camera from the other side.

The Chamber wall of the sample chamber is in a further embodiment instead of the previous expensive silicon chip with elaborately manufactured separate Heating and sensor structures of a specially designed flexible Printed circuit board formed, the subject of another invention of the Applicant is. As usual, this consists of a very thin plastic film (e.g., 0.1 mm thick, polyimide, or the like) with one or two superimposed Copper layers. In the copper sample chamber facing away from the sample space a meandering structure from very thin Conductor tracks etched, the second layer on the sample chamber side is a continuous copper layer for better heat distribution (this may also be omitted).

By A specially developed electronics will meander the meander as well as heating also used as a sensor and targeted after a calibration heated certain temperatures.

The flexible circuit board is much cheaper than the Si chip, With it you can achieve even better cooling rates and you can against the pressed Cooling plunger still Continue to heat up at the same time less heating of the cooling plunger behind the pressure surface.

In addition, will significantly less power for the pestle to Image acquisition needed, whereby the drive can be made smaller and cheaper.

Claims (19)

Cooling stamp device for controlling a temperature profile in a process chamber, in particular for reaction chambers of an assay processor cartridge, consisting of - a cooling stamp ( 12 ), a substantially insulated metal body, which is mounted linearly movable with respect to the surface to be cooled of the process space, - a cooling piston surface for receiving a linear drive for the purpose of a controllable forward and backward movement and - a to the cooling stamp ( 12 ) arranged Peltier element ( 18 ) with heat sink ( 19 ) and fans ( 20 ). Cooling stamp device for controlling a temperature profile in a process chamber according to claim 1, characterized in that the cooling stamp ( 12 ) consists of two mutually displaceable parts a) and b), wherein a) a cooling die with a central bore ( 11 ) and laterally dissipating cooling surface for attaching the Peltier element is and b) one in the central bore ( 13 ) to the cooling punch movably arranged pin-shaped secondary tappets ( 14 ), which by a screw ( 15 ) is represented. Cooling stamp device for controlling a temperature profile in a process chamber according to claims 1 and / or 2, characterized in that an adapter connected to the linear drive ( 16 ) is arranged so that it has a suitable spring ( 17 ) against the cooling stamp ( 12 ) can exert a controllable pressure. Cooling stamp device for controlling a Temperature profile in a process chamber according to at least one of the preceding claims, characterized in that suitable as a linear drive suitable stepper motors or servo geared motors with spindle or worm gear, linear stepper motors, piezolinear motors, motor with pinion and rack, solenoid, rotary magnet, voice coil magnet or motor Cam can be used. Cooling piston device for controlling a temperature profile in a process room at least one of the preceding claims, characterized that the cooled surfaces of the cooling stamp are surrounded with a suitable insulating material. Cooling piston device for controlling a temperature profile in a process room at least one of the preceding claims, characterized that the cooling stamp made of aluminum or copper or another good heat-conducting Material exists. Cooling piston device for controlling a temperature profile in a process room Claim 6, characterized in that the cooling stamp an aluminum cooling die made of non-anodized or anodized aluminum. Cooling stamp device for controlling a temperature profile in a process chamber according to at least one of the preceding claims, characterized in that for temperature control at least one suitable sensor device ( 22 ) and heat conducting elements ( 24 ) are arranged between the cooling punch and Peltier element. Cooling stamp device for controlling a temperature profile in a process chamber according to claim 8, characterized in that the sensor device ( 22 ) is connected by a thermal adhesive or pressed or screwed with a suitable connection mechanism. Method for controlling a temperature profile in a process space, in particular for reaction chambers of an assay processor cartridge, characterized in that a cooling stamp ( 12 ), consisting of a substantially insulated metal body, equipped with two mutually displaceable parts a) and b), wherein a) a cooling die with a central bore ( 11 ) and laterally dissipating cooling surface for attaching a Peltier elementes and b) one in the central bore ( 13 ) to the cooling punch movably arranged pin-shaped secondary tappets ( 14 ), which is replaced by a screw ( 15 ), contains, via a laterally disposed cooling surface by a thereto with heat sink ( 19 ) and fans ( 20 ) equipped Peltier element ( 18 ), is controlled by means of Peltier controller to a desired temperature, linearly moved by a linear drive for the purpose of a controllable back and forth movement to the AP cartridge and thereby positioned so that the target temperature of the process chamber is achieved by a defined cooling punch drive and defined Kühlstempelandruck. Method for controlling a temperature profile in a process space, in particular for reaction chambers of an assay processor cartridge Claim 10, characterized in that the measurement of the actual temperature the process space is carried out by a suitable temperature sensor and via a suitable control mechanism causes the movement of the cooling punch on the punch drive. Method for controlling a temperature profile in a process space, in particular for reaction chambers of an assay processor cartridge the claims 10 to 11, characterized in that the heating of the process chamber and the drive of the cooling stamp be controlled by the same control element. Method for controlling a temperature profile in a process space, in particular for reaction chambers of an assay processor cartridge the claims 10 to 12, characterized in that the temperature of the cooling stamp constant is held. Method for controlling a temperature profile in a process space, in particular for reaction chambers of an assay processor cartridge the claims 10 to 13, characterized in that the linear drive in the second position only the cooling plunger over a suitable spring positioned against the chamber wall of the process space, until the target temperature is reached. Method for controlling a temperature profile in a process space, in particular for reaction chambers of an assay processor cartridge, according to claims 10 to 14, characterized in that in a third position, the center hole ( 13 ) arranged pin-shaped secondary tappets ( 14 ) moved so far in the direction of the process space until it is positioned with much more force than the cooling die centric and / or punctiform against the chamber wall. Method for controlling a temperature profile in a process space, in particular for reaction chambers of an assay processor cartridge, according to claims 10 to 15, characterized gekenn records that the linear drive is preferably carried out by suitable stepper motors or servo geared motors with spindle or worm gear, linear stepper motors, piezolinear motors, motor with pinion and rack, solenoid, rotary magnet, voice coil magnet or motor with cam. Method for controlling a temperature profile in a process chamber according to claim 16, characterized in that For lifting and rotary magnets, the speed of the armature is damped. Method for controlling a temperature profile in a process space, in particular for reaction chambers of an assay processor cartridge at least one of the claims 10 to 17, characterized in that the tasks of heating and Sensor structures of the process room through a flexible printed circuit board he follows. Method for controlling a temperature profile in a process space, in particular for reaction chambers of an assay processor cartridge at least one of the claims 10 to 17, characterized in that the linear drive after a Reference measurement A is programmed so that it has a cooling position B and can assume a position C that lies between B and A.