JP2008067622A - Cell trapping device and temperature control method for the cell trapping device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a cell trapping device which accurately measures the temperature in the vicinity of cells and controls the temperature with low power consumption, and to provide a temperature control method of the cell trapping device. <P>SOLUTION: The cell trapping device is equipped with a cell trapping substrate which sucks a culture solution containing cells and traps the cells into suction holes, heater parts which are arranged on the surface of the substrate and heat the culture solution by control voltage, temperature sensor parts which are arranged on the surface of the substrate and detect the temperature of the culture solution and a control part which controls the control voltage, based on the detected temperature and the target temperature of the culture solution. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a cell trapping device that accurately measures the temperature in the vicinity of a cell and controls it with low power consumption, and a temperature control method for the cell trapping device.

  In recent years, attention has been focused on methods for improving diseases by injecting genes into cells to treat diseases caused by genetic causes. As a result, the role of genes can be elucidated, and tailor-made medicine that performs medications and treatments tailored to individual genetic characteristics has become possible.

  As a method of injecting genes into cells, an electrical method using high-pressure pulses (electroporation method), a chemical method using endocytosis or membrane fusion (lipofection method), or a machine using a fine needle A typical method (microinjection method) is known. However, the electrical method has a large damage to cells, and the chemical method has poor gene injection efficiency. For these reasons, mechanical methods are now considered the safest and most efficient injection method.

  In order to perform the mechanical gene injection, it is necessary to mechanically capture cells.

  FIG. 1 shows an example of a micro semiconductor substrate (hereinafter referred to as a cell capture chip) that performs cell capture using a silicon (hereinafter referred to as Si) material by a conventional mechanical method. FIG. b) shows a cross-sectional view.

  In FIG. 1, 101 is a cell trapping chip, 102 is a surface Si layer, 103 is a cell trapping hole, 104 is a cell, 105 is an insulating layer, 106 is a Si substrate, and 107 is a protective film.

  The cell trapping chip 101 includes a plurality of cell trapping holes 103 that are smaller than the diameter of the cell 104, and a cell culture hole (not shown) containing the cells 104 is sucked from the lower part of the cell trapping hole 103, thereby The cells 104 are adsorbed and fixed at the entrance portion.

  Other means for immobilizing cells include applying a high frequency between the electrodes and immobilizing the cells on the microelectrodes on the cell capture chip using the force of the electric field acting on the cells, or having no affinity for the cells For example, a cell affinity substance is arranged on a cell capture chip, and cells are fixed on the cell affinity substance. By inserting a fine injection needle (capillary) into each of the cells fixed by these means and sequentially injecting the substance, a large amount of injection processing into the cells can be performed.

  In the cell capture and injection process, temperature control near the cells is an important condition.

As a conventional temperature control of a container for culturing cells, there is a method of controlling the temperature by providing heat generating flat plates (hereinafter referred to as plates) on the top and bottom of the container and attaching a temperature sensor to the plate (see Patent Document 1).
JP-A-10-28576

  In FIG. 1, when the cell trapping chip 101 is exposed to room temperature without temperature control, the cells 104 are mixed and injected into the culture solution on the cell trapping chip 101 and then trapped in the cell trapping hole 103, and the substance is collected by the injection needle. Until the cell 104 is completely injected into the cell 104, the periphery of the cell 104 and the cell 104 change to room temperature, and the temperature environment suitable for the cell cannot be kept constant.

  In addition, the temperature cannot be controlled when observing the optimum temperature for injecting the substance into the cell or the reaction to the temperature of the injected cell.

  In order to control the temperature, when the above-described temperature control using the plate is used, a very large heat capacity is required to keep the entire culture vessel at a constant temperature. Furthermore, since there are many heat sources such as a table for controlling the position of the culture vessel and irradiation light for illuminating the cells, it is difficult to control the temperature with low power consumption.

  In addition, since temperature control by the plate is based on temperature measurement at a position away from the cell, there is a possibility that the temperature in the vicinity of the cell may be different, and what effect the temperature in the vicinity of the cell has when the substance is injected into the cell. I could not know exactly.

  That is, although the temperature control method in the vicinity of the cells using the plate described above can control the average temperature of the entire culture vessel, there is a problem with the temperature control in the cell capture and injection processes.

  Accordingly, one of the objects of the present invention is to provide a cell trapping device and a temperature control method for the cell trapping device that accurately measure and control the temperature in the vicinity of the trapped cells.

  In addition, it is a result derived | led-out by each structure shown in the best form for implementing invention mentioned later not only the said objective, Comprising: The effect which is not acquired by the prior art is also one of the other objectives of this invention. Can be positioned.

(1) In the present invention, a cell trapping substrate that sucks a culture solution containing cells and traps the cells in a suction hole, a heater unit that is disposed on the substrate surface and heats the culture solution by a control voltage, and the substrate A cell comprising: a temperature sensor unit disposed on a surface for detecting the temperature of the culture solution; and a control unit for controlling the control voltage based on the detected temperature and a target temperature of the culture solution. Use a capture device.
(2) In addition, a plurality of sets of the heater unit and the temperature sensor unit are arranged on the surface of the substrate on the side filled with the culture solution, and the temperature is controlled based on different target temperatures. The cell capture device according to claim 1 is used.
(3) In addition, a plurality of sets of the heater unit and the temperature sensor unit are arranged in contact with the culture solution on the back surface of the substrate and on the side for sucking the culture solution, and the temperature is controlled based on different target temperatures. The cell capturing device according to claim 1 is used.
(4) The cell trapping device according to claim 1, wherein the heater part and the temperature sensor part are covered with an insulating protective film.
(5) In the present invention, a step of aspirating a culture solution containing cells to capture the cells in the suction holes of the cell capture substrate, and applying a control voltage to a heater portion disposed on the surface of the substrate, A step of heating; a step of detecting a temperature of the culture medium by a temperature sensor unit disposed on a surface of the substrate; and a step of controlling the control voltage based on the detected temperature and a target temperature of the culture medium And a temperature control method for a cell trapping device.

  INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide a cell capturing device that accurately measures the temperature in the vicinity of a cell and controls it with low power consumption and a temperature control method for the cell capturing device.

Hereinafter, embodiments of the present invention will be described with reference to the drawings.
(Example 1)
In Example 1, a heater and a temperature sensor are arranged on the surface side of the cell trapping chip to control the temperature in the vicinity of the cell.

  FIG. 2 shows the overall configuration of the cell trapping apparatus in Example 1.

  In FIG. 2, 200 is a cell trapping device, 201 is a cell trapping chip, 202 is a temperature sensor wire, 203 is a heater wire, 204 is a control unit, 205 is a support, 206 is an injection tube, 207 is a suction tube, and 208 is a petri dish. 209, a culture solution containing cells, 210 a culture solution, 211 a stage, 212 a fine injection needle, 213 a light source, and 214 a microscope.

  The operation of the cell trapping apparatus 200 using the cell trapping chip 201 will be described.

  As shown in FIG. 2, the cell trapping chip 201 is mounted on a support table 205 in a petri dish 208, and a lower part of the support table 205 is filled with a culture solution 209 containing cells injected into the upper part of the support table 205. The culture solution 210 is sucked by the suction tube 207.

  The cell trapping chip 201 has a structure having fine cell capture holes penetrating vertically, and the cells contained in the upper culture solution 209 are captured at the entrance of the cell capture hole by suction of the culture solution 210. The trapped cells are held in the cell trapping hole by continuous suction with the suction tube 207.

  While observing the microscope 214 by irradiation from the light source 213, the stage 211 is moved back and forth and left and right, the position of the cell 311 is set by adjusting the position of the petri dish 208 on the stage 211, and a predetermined substance is introduced by the fine injection needle 212. Injected.

  FIG. 3 shows the structure of the cell trapping chip 201 in Example 1, (a) of FIG. 3 is a surface view, and (b) is a cross-sectional view.

  The cell trapping chip 201 has a structure using an SOI (Silicon on Insulator) substrate made of Si.

In FIG. 3, the cell trapping chip 201 is numbered the same as in FIG. 2, 302 is a surface Si layer, 303 is a heater (platinum: Pt), 304 is a heater (platinum: Pt), 305 is a temperature sensor (platinum). : Pt), 306 is a temperature sensor (platinum: Pt), 307 is a cell trapping hole, 308 is a protective film (silicon nitride: Si ₃ N ₄ ), 309 is an insulating layer (silicon oxide: SiO ₂ ), and 310 is a Si substrate. Reference numeral 311 denotes a cell.

  The shapes of the heaters 303 and 304 and the temperature sensors 305 and 306 are not limited to the shapes shown in FIG. 3, and the arrangement order of the heaters 303 and 304 and the temperature sensors 305 and 306 is not limited to that shown in FIG. However, the object of the present invention can be achieved as long as it is in the vicinity of the cell trapping hole 307.

  4 shows a temperature profile of the cell trapping chip, FIG. 4A shows a temperature profile, and FIG. 4B shows a cross-sectional view of the corresponding cell trapping chip.

4, the same components as those in FIG. 3 are denoted by the same reference numerals, T ₁ indicates a high temperature (for example, + 40 ° C.), and T ₂ indicates a low temperature (for example, + 30 ° C.).

  The operation of the cell trapping chip 201 will be described with reference to FIG. 2, FIG. 3, and FIG.

  The culture solution 209 containing the cells 311 is injected onto the surface Si layer 302 of the cell trapping chip 201 by the injection tube 206. The culture solution 210 under the cell capture hole 307 is sucked by the suction tube 207, and the cell 311 is captured at the entrance of the cell capture hole 307.

  The temperature control of the culture solution 209 containing the cells 311 will be described.

Control unit 204, when measured target temperature above T ₁ by the temperature sensor 305, performs feedback control so as to stop the heating of the heater 303 and measures the target temperature T ₁ of less subjected to feedback control so as to perform the heating of the heater 303, near the temperature sensor 305 performs temperature control by the on / off such that the temperature T _1.

  Moreover, high-precision continuous temperature control by feedback control is performed as follows, for example.

The resistance value of platinum Pt of the temperature sensor 305 increases with temperature. The control unit 204 internally connects a stable resistor Rs maintained at a predetermined temperature in series with the temperature sensor 305, and applies a stable constant voltage V ₀ across the resistor Rs and the temperature sensor 305.

The temperature sensor 305 can detect a voltage change at both ends of the platinum Pt because the resistance value changes due to a temperature change in the vicinity and the voltage dividing ratio of the constant voltage V ₀ changes.

That is, the voltage between the terminals of the temperature sensor 305 is obtained as the measurement voltage V _m corresponding to the temperature T _m of the temperature sensor 305. The control unit 204 obtains a voltage difference e _x = (V ₁ −V _m ) between the reference voltage V ₁ set in advance corresponding to the target temperature T ₁ and the measured voltage V _m . When e _x > 0, since the temperature T _m of the temperature sensor 305 is lower than the target temperature, the control unit 204 amplifies the voltage difference e _x with an appropriate amplification degree, and uses the heater 303 as a heating voltage corresponding to the temperature difference. To supply. When e _x ≦ 0, since the temperature T _m of the temperature sensor 305 is equal to or higher than the target temperature, the heating voltage to the heater 303 is set to zero. As a result, the temperature T _m of the temperature sensor 305 is controlled to be accurately the target temperature T ₁ .

The control unit 204 when measuring the target temperature T ₂ or more by the temperature sensor 306, performs feedback control so as to stop the heating of the heater 304 and measures the target temperature T ₂ below provides feedback control to perform the heating of the heater 304 , near the temperature sensor 306 is temperature controlled to a temperature T _2.

Also, the control of the temperature T _2, wherein a can be carried out in to accurate continuous temperature control of by the feedback control as well.

By the above operation, the temperature of the culture solution 209 containing cells is maintained at a temperature T ₁ (for example, + 40 ° C.) in the vicinity of the temperature sensor 305 and a temperature T ₂ (for example, + 30 ° C.) in the vicinity of the temperature sensor 306 as shown in FIG. To be kept.

  Since the temperature sensor 305 is disposed close to the heater 303 and the temperature sensor 306 is disposed close to the heater 304, the culture solution 209 near the heater 303 and the culture solution near the heater 304 in FIG. A temperature gradient is generated between 209 and the cells trapped in the cell trapping hole 307 can introduce substances at the culture solution temperature at each position.

  Since the heaters 303 and 304 and the temperature sensors 305 and 306 are provided in the vicinity of the cell trapping hole 307, the temperature of the culture solution 209 in the vicinity of the trapped cell 311 can be controlled extremely accurately.

  Thereby, since the temperature of the cell during the substance injection process is kept constant, the change of the reaction due to the temperature change can be prevented, and the substance injection and observation can be performed in a constant environment. Furthermore, since a temperature gradient can be applied to the culture solution 209 containing the cells 311, different temperatures can be given to the cell group by a single treatment.

  Next, a manufacturing process of the cell trapping chip 201 in the first embodiment will be described with reference to FIG.

5, the same components as those in FIG. 3 are denoted by the same reference numerals, 500 is an SOI substrate, 501 is a photoresist, 502 is a protective film (silicon nitride: Si ₃ N ₄ ), and 503 is a protective film (silicon nitride: Si ₃ N ₄₎ are shown, respectively, process (a) the process of forming the cell capturing hole 307, the process (b) a protective film in the portion to remain in the etching (silicon _nitride: the process of forming the Si ₃ N 4) 502, The process (c) is a process for forming the heaters 303 and 304 and the temperature sensors 305 and 306, the process (d) is a process for removing the back side of the cell trapping hole 307 of the Si substrate 310, and the process (e) is a protective film ( Processes for removing unnecessary portions and 503 of silicon nitride: Si ₃ N ₄ ) 502 are shown.

  With reference to FIG. 5, the process of producing the structure of the cell trapping chip 201 in Example 1 will be described in detail.

  In the process (a) of FIG. 5, in order to form a plurality of cell trapping holes 307 in the surface Si layer 302 of the SOI substrate 500, the SOI substrate 500 is first cleaned, and then the photoresist 501 is applied to the entire surface of the surface Si layer 302. Apply. The material and thickness of the photoresist 501 are not particularly limited.

  The photoresist 501 is exposed using a photomask and a mask aligner, and the resist is removed from the exposed portion by development to form a pattern of the resist 501 from which the cell trapping hole 307 has been removed. Using the pattern of the resist 501 as an etching mask, a through hole 307 is formed using a dry etching apparatus.

In the process (b), a protective film (silicon nitride: for forming the heaters 303 and 304 and the temperature sensors 305 and 306 on the surface of the surface Si layer 302 and for performing selective etching of the Si substrate 310 in a later process. Si ₃ N ₄ ) 502 and 503 are coated. The protective films (silicon nitride: Si ₃ N ₄ ) 502 and 503 may be silicon oxide (SiO ₂ ) films, and are formed by a CVD (Chemical Vapor Deposition) method in order to protect the side surfaces in the cell trapping holes 307. Is desirable.

A pattern of a protective film (silicon nitride: Si ₃ N ₄ ) 503 is formed on the back surface of the Si substrate 310 by using a photoresist and exposure and etching as a mask for performing selective etching of the Si substrate 310 in a later process. .

In the process (c), platinum (hereinafter referred to as Pt) is formed on the surface of the surface Si layer 302 as a material for the heaters 303 and 304 and the temperature sensors 305 and 306. In order to improve the adhesion to the protective film (silicon nitride: Si ₃ N ₄ ) 502, titanium (hereinafter referred to as Ti) is formed before the platinum film is formed (not shown). The film forming method is performed using a vapor deposition method or a sputtering method.

  As a film formation method of the heaters 303 and 304 and the temperature sensors 305 and 306, first, a Pt film is formed on the entire surface of the Ti film on the surface Si layer 302, and then the heaters 303 and 304 and the temperature are formed by photoresist, exposure and etching. First, a resist is formed on the entire surface of the Ti film on the surface Si layer 302, the resist is removed by drawing with an electron beam, and a Pt film is formed on the removed portion. Either lift-off method may be used.

  The material of the heater and the temperature sensor is Pt, but is not particularly limited.

  In the process (d), the Si substrate 310 is etched.

  The surface shown in FIG. 5D is produced by protecting the surface of the surface Si layer 302 and anisotropically etching the Si substrate 310 with an alkaline aqueous solution or the like from the back surface. Note that dry etching may be used for silicon etching of the Si substrate 310.

In the process (e), an unnecessary portion of the protective film (silicon nitride: Si ₃ N ₄ ) 502 on the surface of the surface Si layer 302 and a protective film (silicon nitride: Si ₃ N ₄ ) 503 on the back surface of the Si substrate 310, The insulating layer 309 in the vicinity of the cell trapping hole 307 on the back surface of the surface Si layer 302 is removed, and the shape shown in FIG.
(Example 2)
In Example 2, a heater and a temperature sensor are arranged on the back side of the cell trapping chip to control the temperature in the vicinity of the cell.

 In the first embodiment, since the heaters 303 and 304 and the temperature sensors 305 and 306 directly touch the culture solution 209 containing the cells 311, electrical stimulation or thermal stimulation may be directly transmitted to the cells and affected.

  In order to alleviate these stimuli, in Example 2, a heater and a temperature sensor are arranged on the back side of the cell trapping chip 201 that is the side for sucking the culture solution 209 containing the cells 311.

  FIG. 6 shows the structure of the cell trapping chip according to Example 2.

  FIG. 6A shows a back view and FIG. 6B shows a cross-sectional view.

  In Example 2, as in Example 1, the cell trapping chip has a structure using an SOI (Silicon on Insulator) substrate made of Si.

In FIG. 6, the same components as those in FIG. 3 are denoted by the same reference numerals, 601 is a cell trapping chip, 602 is a protective film (silicon nitride: Si ₃ N ₄ ), 603 is a heater (platinum: Pt), and 604 is A heater (platinum: Pt), 605 indicates a temperature sensor (platinum: Pt), and 606 indicates a temperature sensor (platinum: Pt).

  The shapes of the heaters 603 and 604 and the temperature sensors 605 and 606 are not limited to the shapes shown in FIG. 3, and the arrangement order of the heaters 603 and 604 and the temperature sensors 605 and 606 is not limited to that shown in FIG. However, the object of the present invention can be achieved as long as it is in the vicinity of the cell trapping hole 307.

  In the second embodiment, the operation of the cell trapping chip 601 for capturing the cells 311 and the temperature control operation are the same as the operation of the cell trapping chip 201 in the first embodiment.

  The overall configuration of the cell trapping apparatus in the second embodiment is the same as that of the second embodiment except that the cell trapping chip 201 in FIG.

  The temperature profile of the cell trapping chip 601 in Example 2 is the same as that in FIG.

  Further, regarding the process of manufacturing the cell trapping chip 601, it can be manufactured using the same process as that of FIG. 5 showing the process of manufacturing the cell trapping chip 201 in the first embodiment.

  The second embodiment is different from the first embodiment in that the heaters 603 and 604 and the temperature sensors 605 and 606 do not directly touch the culture solution 209 containing the cells 311, and thus the electrical and thermal direct by the heater and the temperature sensor are not used. It is not to be stimulated.

  Moreover, another example which arrange | positions a heater and a temperature sensor in the back surface of a cell capture chip | tip is shown in FIG.

  FIG. 7A is a back view of the cell trapping chip, and FIG. 7B is a cross-sectional view of the cell trapping chip.

  7, the same components as those in FIG. 6 are denoted by the same reference numerals, 701 is a cell trapping chip, 703 is a heater (platinum: Pt), 704 is a heater (platinum: Pt), and 705 is a temperature sensor (platinum: Pt) and 706 indicate temperature sensors (platinum: Pt), respectively.

  The cell trapping and temperature control operations of the second embodiment shown in FIG. 7 are the same as the operations in FIG. The cell capture chip 701 in FIG. 7 is different from the cell capture chip 601 in FIG. 6 because the sensors 705 and 706 and the heaters 703 and 704 in FIG. 7 are arranged on the back surface of the surface Si layer 302, compared to FIG. Since temperature measurement and heating can be performed in the vicinity of the captured cell 311, temperature control with higher accuracy and quick response is possible.

Also in the structure in FIG. 7, since the heaters 703 and 704 and the temperature sensors 705 and 706 do not directly touch the culture solution 209 containing the cells 311, they are not subjected to direct electrical and thermal stimulation by the heater or the temperature sensor. There is.
(Example 3)
In Example 3, the heater and the temperature sensor arranged on the side of the cell trapping chip are covered with a protective film (silicon nitride: Si ₃ N ₄ ) to control the temperature in the vicinity of the cell.

 In the first embodiment, since the heaters 303 and 304 and the temperature sensors 305 and 306 directly touch the culture solution 209 containing the cells 311, electrical stimulation or thermal stimulation may be directly transmitted to the cells 311 and affected.

In order to alleviate these stimuli, in Example 3, the heater and the temperature sensor are covered with a protective film (silicon nitride: Si ₃ N ₄ ) so that the cells do not directly touch the heater and the sensor.

  FIG. 8 shows the structure of the cell trapping chip according to Example 3.

  FIG. 8A shows a surface view and FIG. 8B shows a cross-sectional view.

  In Example 3, as in Example 1, the cell trapping chip 801 has a structure using an SOI (Silicon on Insulator) substrate made of Si.

In FIG. 8, the same components as those in FIG. 3 are denoted by the same reference numerals, 801 is a cell trapping chip, 802 is a protective film (silicon nitride: Si ₃ N ₄ ), 803 is a heater (platinum: Pt), and 803a is Heater pad, 803b heater pad, 804 heater (platinum: Pt), 804a heater pad, 804b heater pad, 805 temperature sensor (platinum: Pt), 805a temperature sensor pad, 805b temperature sensor pad, 806 Is a temperature sensor (platinum: Pt), 806a is a temperature sensor pad, and 806b is a temperature sensor pad.

  The shapes of the heaters 803 and 804 and the temperature sensors 805 and 806 are not limited to the shapes shown in FIG. 8, and the arrangement order of the heaters 803 and 804 and the temperature sensors 805 and 806 is not limited to that shown in FIG. However, the object of the present invention can be achieved as long as it is in the vicinity of the cell trapping hole 307.

  In the third embodiment, the operation of the cell trapping chip 801 for capturing the cells 311 and the temperature control operation are the same as the operation of the cell trapping chip 201 in the first embodiment.

  The overall configuration of the cell trapping apparatus in the third embodiment is the same as that of the third embodiment except that the cell trapping chip 201 in FIG.

  The temperature profile of the cell trapping chip 601 in Example 2 is the same as that in FIG.

  Moreover, regarding the manufacturing process of the cell trapping chip 801, it can be manufactured by using the same process as FIG. 5 showing the manufacturing process of the cell trapping chip 201 in the first embodiment.

However, the heaters 803 and 804 and the temperature sensors 805 and 806 arranged on the surface side of the cell trapping chip 801 are covered with a protective film (silicon nitride: Si ₃ N ₄ ) 802 and the heater pads (external connection electrodes) ) 803a, 803b, 804a, 804b and temperature sensor pads 805a, 805b, 806a, 806b are subjected to a process not covering the protective film.

  The third embodiment is different from the first embodiment in that the heaters 803 and 804 and the temperature sensors 805 and 806 do not directly touch the culture solution 209 containing the cells 311, so that the electrical and thermal direct by the heater and the temperature sensor are not. It is not to be stimulated.

It is a figure which shows a cell capture chip | tip (conventional example). It is a figure which shows a cell capture apparatus structure (Example 1). It is a figure which shows the structure (Example 1) of a cell trapping chip. It is a figure which shows the temperature profile (Example 1) of a cell capture chip | tip. It is a figure which shows the preparation process (Example 1) of a cell capture chip | tip. It is a figure which shows the structure (Example 2) of a cell trapping chip. It is a figure which shows the structure (another example of Example 2) of a cell capture chip | tip. It is a figure which shows the structure (Example 3) of a cell trapping chip.

Explanation of symbols

DESCRIPTION OF SYMBOLS 101 Cell capture chip 102 Surface Si layer 103 Cell capture hole 104 Cell 105 Insulating layer 106 Si substrate 107 Protective film 200 Cell capture device 201 Cell capture chip 202 Temperature sensor line 203 Heater line 204 Control part 205 Support stand 206 Injection tube 207 Suction tube 208 Petri dish 209 Culture solution containing cells 210 Culture solution 211 Stage 212 Fine injection needle 213 Light source 214 Microscope 302 Surface Si layer 303 Heater (platinum: Pt)
304 Heater (Platinum: Pt)
305 Temperature sensor (Platinum: Pt)
306 Temperature sensor (Platinum: Pt)
307 Cell trapping hole 308 Protective film (silicon nitride: Si ₃ N ₄ )
309 Insulating layer (silicon oxide: SiO ₂ )
310 Si substrate 311 Cell 501 Photoresist 502 Protective film (silicon nitride: Si ₃ N ₄ )
503 Protective film (silicon nitride: Si ₃ N ₄ )
601 Cell trapping chip 602 Protective film (silicon nitride: Si ₃ N ₄ )
603 Heater (Platinum: Pt)
604 Heater (Platinum: Pt)
605 Temperature sensor (Platinum: Pt)
606 Temperature sensor (Platinum: Pt)
701 Cell capture chip 703 Heater (Platinum: Pt)
704 Heater (Platinum: Pt)
705 Temperature sensor (Platinum: Pt)
706 Temperature sensor (Platinum: Pt)
801 Cell trapping chip 802 Protective film (silicon nitride: Si ₃ N ₄ )
803 Heater (Platinum: Pt)
803a Heater pad 803b Heater pad 804 Heater (platinum: Pt)
804a Heater pad 804b Heater pad 805 Temperature sensor (platinum: Pt)
805a Temperature sensor pad 805b Temperature sensor pad 806 Temperature sensor (platinum: Pt)
806a Temperature sensor pad 806b Temperature sensor pad

Claims (5)

A cell-capturing substrate for sucking a culture solution containing cells and capturing the cells in the suction holes;
A heater unit arranged on the surface of the substrate for heating the culture solution by a control voltage;
A temperature sensor unit disposed on the substrate surface for detecting the temperature of the culture solution;
A control unit for controlling the control voltage based on the detected temperature and a target temperature of the culture medium;
A cell capturing device comprising:   The said heater part and the said temperature sensor part are arrange | positioned in multiple numbers on the surface of the said board | substrate of the side filled with the said culture solution, and are temperature-controlled based on different target temperature, respectively. Cell capture device.   The heater unit and the temperature sensor unit are arranged in contact with the culture medium on the back surface of the substrate and sucking the culture medium, and the temperature is controlled based on different target temperatures. The cell capturing device according to claim 1.   The cell capture device according to claim 1, wherein the heater unit and the temperature sensor unit are covered with a protective film made of an insulating material. Aspirating a culture solution containing cells to capture the cells in the suction holes of the cell capture substrate;
Heating the culture medium by applying a control voltage to each of the plurality of heaters disposed on the surface of the substrate;
Detecting the temperature of the culture solution by a plurality of temperature sensor units arranged on the surface of the substrate;
Controlling each of the control voltages based on the plurality of detected temperatures and a plurality of target temperatures of the culture medium;
A temperature control method for a cell trapping device comprising:

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