JP2019140914A - Heat transfer plate, well plate unit and cell peeling device - Google Patents

Abstract

To collect a cell in a non-invasive condition for a cell.SOLUTION: The invention relates to a heat transfer plate on which, a container having a plurality of wells for storing a cell, is mounted, which comprises a body part having a heat transfer coefficient higher than that of the container, and contacting at least a part of an outside bottom surface of each of the plurality of wells, and a plurality of transparent open parts penetrating the body part on a lower side of at least parts of inside bottom surfaces of the plurality of wells, and relates to a cell peeling device having the heat transfer plate.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a heat transfer plate, a well plate unit, and a cell peeling device.

A method for recovering a target cell by culturing and differentiating universal cells such as iPS cells is known. However, since there is a relatively low probability that iPS cell establishment and differentiation induction after establishment will be successful, it is necessary to select and recover optimally selected cells in terms of differentiation ability and safety from the cultured cells. A method is known in which cells are cultured on a collagen gel containing gold nanoparticles, and the cells are irradiated with light to solderize the collagen gel in the light irradiation region, and the cells contained in the light irradiation region are detached and recovered. (For example, refer to Patent Documents 1 and 2).
Patent Document 1 Japanese Patent Application Laid-Open No. 2012-39947

  However, according to the conventional method, a high light irradiation intensity and / or a long light irradiation time are required, and the cell is highly invasive.

  In the first aspect of the present invention, there is provided a heat transfer plate on which a container having a plurality of wells in which cells are stored is placed, and has a higher thermal conductivity than the container, and is disposed outside each of the plurality of wells. Provided is a heat transfer plate having a main body part contacting at least a part of the bottom surface and a plurality of transparent through parts penetrating the main body part below at least a part of the bottom surface inside each of the plurality of wells.

  In the second aspect of the present invention, a container having a plurality of wells, a heat transfer plate on which the container is placed, a heater disposed in contact with the heat transfer plate, and a target region in the well are irradiated with light. A heat radiating portion, and the heat transfer plate has a higher thermal conductivity than the container, a main body portion in contact with at least a part of an outer bottom surface of each of the plurality of wells, and an inner bottom surface of each of the plurality of wells There is provided a cell detaching device having a plurality of transparent penetrating parts penetrating through a main body part below at least a part of.

  In the third aspect of the present invention, a container having a plurality of wells in which cells are stored, and a heat transfer plate on which the container is placed, the heat transfer plate has higher thermal conductivity than the container, A main body that is in contact with at least a part of the outer bottom surface of each of the plurality of wells; and a plurality of transparent through portions that penetrate the main body part below at least a part of the inner bottom surface of each of the plurality of wells. A well plate unit is provided.

  The summary of the invention does not enumerate all the features of the present invention. In addition, a sub-combination of these feature groups can also be an invention.

An example of the structure of the cell peeling apparatus 10 of this embodiment is shown. An example of the structure of the heat-transfer plate 200 of this embodiment is shown. An example of the structure of the well 110 of this embodiment is shown. An example of the external appearance of the container 100 of this embodiment is shown. An example of the external appearance of the heat-transfer plate 200 of this embodiment corresponding to FIG. 4 is shown. An example of the well plate unit of this embodiment is shown. An example of the processing flow of the cell peeling apparatus 10 of this embodiment is shown. An example of the effect of the cell peeling method of this embodiment is shown.

  Hereinafter, the present invention will be described through embodiments of the invention, but the following embodiments do not limit the invention according to the claims. In addition, not all the combinations of features described in the embodiments are essential for the solving means of the invention.

  FIG. 1 shows an example of the configuration of the cell peeling apparatus 10 of the present embodiment. The cell peeling apparatus 10 selectively peels and collects target cells using light irradiation. The cell peeling device 10 includes a container 100, a heat transfer plate 200, a heater 300, a driving unit 350, a light irradiation unit 400, an optical member 500, a mirror 600, a lens 700, an observation unit 800, and a cell acquisition unit 900.

  The container 100 stores cells to be peeled off. The container 100 has a plurality of wells 110 in which cells are stored, and the cells are stored in each of the wells 110. For example, the plurality of wells 110 are a plurality of indentations provided on the upper plate of the container 100. The container 100 is formed of a transparent material such as resin or glass. The container 100 stores a medium whose structure is changed by heating in each well 110, and holds cells on the medium. As an example, the medium may include a gel that forms a sol by heating and gold fine particles.

  The heat transfer plate 200 has the container 100 placed thereon and transfers heat from the heater 300 to the container 100. The heat transfer plate 200 has a main body part 210 and a plurality of through parts 220, and mainly conducts heat in the main body part 210 and transmits light in the through part 220. The main body 210 may have a higher thermal conductivity than the container 100. The plurality of through portions 220 are respectively provided at positions corresponding to some or all of the plurality of wells 110 in the main body 210. For example, the penetration part 220 may be provided as a power of 2 (2, 4, 8, 16, 32, 64, etc.). Further, the plurality of through portions 220 may be provided at equal intervals in the main surface of the heat transfer plate 200. Thereby, the heat transfer plate 200 enables light irradiation to the cells in the well 110 while ensuring that the main body 210 contacts the container 100 and heat conduction to the container 100.

  The main body 210 may be made of a metal having good thermal conductivity. For example, the main body 210 is made of aluminum (for example, the thermal conductivity as a bulk is 226 to 237 W / mK: the values in parentheses indicate the thermal conductivity as a metal bulk, and the unit is W / mK. ), Gold (318), silver (429), copper (386-402), iron (72-80.4), tungsten (169-176), and alloys containing parts thereof. Further, at least a part of the main body 210 may be formed of a non-metallic material such as graphite (for example, thermal conductivity 200 to 2000 W / mK) having good thermal conductivity. Further, the main body 210 may be made of a resin having a higher thermal conductivity as a bulk than the container 100. For example, acrylonitrile / styrene resin (for example, the thermal conductivity as a bulk is 0.15 to 0.16 W / mK: the values in parentheses indicate the thermal conductivity as the bulk of the resin, and the unit is W / mK. ), Acrylonitrile / butadiene / styrene resin (0.19 to 0.36), epoxy resin (0.3), ethylene tetrafluoride / ethylene copolymer (0.25), ethylene tetrafluoride / hexafluoro Propylene copolymer (0.25), polyamide 6 (0.25-0.43), polyamide 66 (0.24-0.43), polyamide 610 (0.22), polybutylene terephthalate (0.25) ), Polycarbonate (0.19), low density polyethylene (0.33), medium density polyethylene (0.33 to 0.42), high density polyethylene (0.46 to 0.52), Reether ether ketone (0.26), polyether sulfone (0.18 to 0.24), polyethylene terephthalate (0.31), phenol resin (0.13 to 0.25), tetrafluoroethylene perfluoro Vinyl ether copolymer (0.25), polyimide (0.28 to 0.34), polymethyl methacrylate (0.17 to 0.25), polyacetal (0.23), polypropylene (0.125), polyphenylene sulfide (0.29), polyphenylene oxide (0.19), polystyrene (0.10-0.14), polysulfone (0.12-0.16), tetrafluoroethylene resin (0.25), polyvinyl chloride (0.13-0.29), polyvinylidene chloride (0.13), polyvinylidene fluoride (0.13), polytrifluoride Ethylene (0.19-0.29), polytetrafluoroethylene (0.25), polyoxymethylene (0.23-0.29), cellulose acetate (0.20), cellulose acetate butyrate (0 0.17 to 0.33), unsaturated polyester resin (0.17 to 0.21), polyurethane resin (0.21), and silicone resin (0.15 to 0.17), etc. One or more resins having higher thermal conductivity may be selected and used for at least a part of the main body 210. For the container 100, for example, one or more resins having a lower thermal conductivity as a bulk than the main body 210 may be selected and used from the above-described resins. In particular, the main body 210 is preferably formed of aluminum from the viewpoint of thermal conductivity, workability, cost, and the like. When the container 100 is mounted, the main body 210 may be in contact with at least a part of the outer bottom surface of each of the plurality of wells 110. For example, the main body 210 may be disposed so as to cover at least the bottom surfaces of the outer walls of the plurality of wells 110 when the container 100 is mounted.

  Each of the plurality of penetrating portions 220 may be an opening provided to transmit light and penetrate the main body portion 210 in the thickness direction. The plurality of through portions 220 may be voids that are not filled with a material. In this case, the through portions 220 are through holes. Instead, the through portion 220 may be filled with a transparent material such as glass or resin having good thermal conductivity (for example, a resin cited as a candidate for the main body portion 210). The penetrating portion 220 transmits at least a part of the wavelength included in the emitted light of the light irradiation unit 400, and preferably transmits visible light. The through portion 220 may be provided below at least a part of the bottom surface inside each of the plurality of wells 110.

  Here, the heat transfer plate 200 may have a shape and a thickness that match the shape of the container 100 so that the container 100 is in close contact with the heat transfer plate 200. For example, when the container 100 has legs that protrude from the bottom surface of the container 100 at the outer edge, the heat transfer plate 200 has a predetermined size (for example, 0.01 to 1 mm) than the length that the legs protrude. It may have a large thickness. Thus, the heat transfer plate 200 can prevent the main body other than the legs of the container 100 from floating from the heat transfer plate 200 while maintaining the heat transfer property. As an example, the heat transfer plate 200 may have a thickness of 1 to 3 mm.

  The heater 300 is disposed in contact with the heat transfer plate 200, generates heat, and transfers the heat to the container 100 through the heat transfer plate 200. The heater 300 also functions as a stage for the container 100 and the heat transfer plate 200. That is, the heater 300 moves the container 100 and the heat transfer plate 200 by moving in the three-dimensional space. In addition, a stage for driving the heater 300 may be separately prepared.

  For example, the rectangular heater 300 may be fitted into a stage whose center is hollowed into a rectangular shape and driven together with the stage. The heater 300 has a light transmission part 310 that transmits light to at least a part of a region where the heat transfer plate 200 is mounted. The heater 300 may have a heat generating device (that is, a transparent heat generating device) in the light transmission part 310, and may transmit the heat generated in the light transmission part 310 to the heat transfer plate 200. Instead, the heater 300 includes an opaque or transparent heat generating device around the light transmitting portion 310, and transfers heat from the heat generating device to the heat transfer plate 200 via the light transmitting portion 310. Also good.

  The driving unit 350 drives the heater 300 and moves in a three-dimensional direction. The driving unit 350 drives the heater 300 (or the stage in which the heater 300 is incorporated) in the main surface direction (XY direction) and the vertical direction (Z direction) of the container 100 and the heater 300 by a power unit such as a motor. You can do it. Moreover, the drive part 350 may rotate the heater 300 within a main surface plane.

  The light irradiation unit 400 irradiates the target region in the well 110 of the container 100 with light. The light irradiation unit 400 irradiates at least one cell to be exfoliated in the well through the penetration part 220 and the light transmission part 310 while heating the container 100 with the heater 300, and thereby the target. The resulting cells may be detached from the medium. The light irradiation part 400 should just irradiate the light which provides the energy which is the extent which the medium fluidized by heating fluidizes, and is a laser light source which radiate | emits a light with a wavelength of 400-1200 nm and an intensity | strength of 0.1-1000 mW. It may be.

  The optical member 500 enters the light emitted from the light irradiation unit 400 and adjusts the optical path, intensity, and / or phase of the incident light beam. For example, the optical member 500 may include a digital mirror device (DMD), a spatial light modulator (SLM), and / or a galvanometer mirror.

  The mirror 600 reflects the light emitted from the light irradiation unit 400 and passing through the optical member 500 toward the container 100. The mirror 600 may transmit light other than the light emitted from the light irradiation unit 400 in order to allow the observation unit 800 to observe the cells in the container 100. For example, the mirror 600 may be a dichroic mirror that reflects light having the wavelength of light emitted from the light irradiation unit 400 and transmits light having other wavelengths.

  The lens 700 is disposed on the optical path between the mirror 600 and the heater 300, enters the light reflected by the mirror 600, and emits the incident light toward the light transmission unit 310. The lens 700 functions as an objective lens that enables the observation unit 800 to observe cells in the container 100.

  The observation unit 800 is disposed on the same optical path as the observation target well 110 and the lens 700, and allows the observer to observe the well 110 in an enlarged manner. For example, the observation unit 800 includes an eyepiece lens and functions as a microscope in combination with the lens 700 to allow the observer to observe the enlarged cells. In addition, for example, the observation unit 800 may include an imaging element and output the enlarged image data in the well 110 that has been imaged to a display device or the like.

  The cell acquisition unit 900 acquires cells detached from the medium by the light irradiated from the light irradiation unit 400. For example, the cell acquisition unit 900 may include a capillary that can suck cells by a capillary effect. In addition, the cell acquisition unit 900 may have a mechanism for recovering detached cells by moving the container 100 over the recovery container and inverting it.

  As described above, the cell detachment apparatus 10 transmits the light emitted from the light irradiation unit 400 in the well 110 of the container 100 through the optical member 500, the mirror 600, the lens 700, the light transmission unit 310, and the penetration unit 220. And irradiate the cells to be detached with light. Here, the cell peeling apparatus 10 performs light irradiation on the cells by the light irradiation unit 400 while heating the container 100 with the heater 300.

  Thereby, the cell peeling apparatus 10 can peel a cell from a medium in a shorter time and weak light irradiation. As a result, the cell peeling device 10 can reduce the invasion to the cells and reliably collect the cells with less damage.

  Moreover, the cell peeling apparatus 10 may include a graphite sheet between the heat transfer plate 200 and the container 100. The container 100 may not be in surface contact with the heat transfer plate 200 due to individual differences in the shape of the container 100, the legs of the container 100 being too long, or the surface of the container 100 being uneven. . Even in such a case, at least a part of the space between the container 100 and the heat transfer plate 200 is filled with a graphite sheet having excellent thermal conductivity and flexibility, so that the heat from the heater 300 can be efficiently transferred to the container. 100. In the case where a graphite sheet is provided, an opening having a shape corresponding to the penetrating portion 220 may be provided in the graphite sheet so as not to obstruct observation by the observation unit 800.

  FIG. 2 shows an example of the configuration of the heat transfer plate 200 of the present embodiment. As shown in the figure, the main body 210 of the heat transfer plate 200 may have a multilayer structure, and the through-hole 220 may be a gap. For example, the main body 210 of the heat transfer plate 200 may include a core layer 212, a surface layer 214, and a surface layer 216.

  As an example, the core layer 212 may be an aluminum plate having a thickness, the surface layer 214 and the surface layer 216 may be graphite sheets, and the core layer 212, the surface layer 214, and the surface layer 216 may be bonded with an adhesive or the like. . Thereby, the heat transfer plate 200 can improve thermal conductivity while maintaining strength.

  FIG. 3 shows an example of the configuration of the well 110 of the present embodiment. Here, the figure which expanded the well containing the cell of peeling object among the some wells 110 of the container 100 is shown. As shown in the figure, during the cell detachment process, the well 110 includes a medium 112, a cell 117, and a culture solution 118.

  The medium 112 includes a gel 114 and gold fine particles 116. The gel 114 may be a gel material whose structure changes when the temperature is equal to or higher than a predetermined temperature (for example, 40 ° C.), and may be, for example, a collagen gel or a gelatin gel. The gold fine particles 116 generate heat by absorbing light having a wavelength emitted from the light irradiation unit 400.

  As a result, the structure of only the gel 114 is changed by the heat generation of the gold fine particles 116 in the vicinity of the region where the light from the light irradiation unit 400 (indicated by the dotted arrow in the drawing) is incident. As a result, the gel 114 is included in the region irradiated with light. Only the cells 117 that are lost lose the scaffold and are detached from the medium 112. The observer may irradiate the target cell 117 with light while observing the inside of the well 110 with the observation unit 800. As a result, the cell acquiring unit 900 can selectively acquire only the target cell 117 by peeling it out of the large number of cells 117 stored in the observation well 110.

  FIG. 4 shows an example of the appearance of the container 100 of the present embodiment. As illustrated, the container 100 may be a microplate in which a plurality of wells 110 are arranged in a matrix. The part other than the part where the well 110 is formed in the container 100 may be filled with the forming material of the container 100. Alternatively, at least a part may be a void.

  FIG. 5 shows an example of the appearance of the heat transfer plate 200 of the present embodiment corresponding to FIG. As shown in the drawing, the heat transfer plate 200 may be a plate provided with a plurality of through portions 220 provided in a matrix corresponding to the wells 110 of FIG. FIG. 5 shows a form in which each of the plurality of openings is provided at equal intervals in the row direction and / or column direction of the matrix and is larger than the bottom surface of the corresponding well.

  This facilitates the alignment of the well 110 and the through portion 220. Alternatively, each of the plurality of openings may be smaller than the bottom surface of the corresponding well 110. In this case, since the area of the main body 210 of the heat transfer plate 200 is increased, the thermal conductivity with respect to the well 110 is improved.

  FIG. 6 shows an example of the well plate unit of the present embodiment. In the present embodiment, the container 100 and the heat transfer plate 200 may be combined to form a well plate unit. The well plate unit may have a form in which the containers 100 are stacked as they are on the heat transfer plate 200 as shown in FIG. 1, but as shown in FIG. The heat transfer plate 200 may be incorporated on the side.

  In this case, the container 100 includes a portion for forming the well 110, an upper plate portion, and a side wall portion. That is, the container 100 has an upper plate provided with a recess for forming the well 110 on the upper side of FIG. 6, but has no plate on the lower side and is a gap, and the through-hole 220 of the heat transfer plate 200 is also provided. It becomes a void.

  Thus, each of the plurality of wells 110 is fitted into the plurality of openings of the corresponding through portion 220. In the present embodiment, each of the plurality of openings is larger than the bottom surface of the corresponding well 110. According to this embodiment, since the heat transfer plate 200 is also heated from the side portion of the container 100, heat can be transferred to the container 100 more efficiently.

  FIG. 7 shows an example of a processing flow of the cell peeling apparatus 10 of the present embodiment. In the present embodiment, the cell peeling device 10 performs the processes of S110 to S190.

  First, in S110, a medium 112 that is fluidized by heating is supplied to a container 100 including a plurality of wells 110 as a cell culture substrate. The medium 112 may include a gel and gold fine particles that are fluidized by heating. For example, as described in Patent Documents 1 and 2, an acetylated collagen peptide-modified dendrimer solution is produced from an acetylated collagen peptide, mixed with a tetrachloroauric (III) acid solution, and then gold ions are reduced. A collagen gel containing gold fine particles is generated, and this may be used as the medium 112 in this embodiment.

  In S110, the medium 112 may be supplied to all or a part of the plurality of wells 110 of the container 100. Further, after the medium 112 is supplied, phosphate buffered saline (PBS (−)) is supplied, and the solution is stored in the well 110 at a predetermined temperature (for example, 37 ° C.) for a predetermined time (for example, 6 hours). By holding this, the medium 112 may be cleaned.

Next, in S120, the cells 117 are fixed to the medium 112. For example, 8.0 × 10 ³ Hela cells may be seeded in the medium 112. Further, in S120, a culture solution 118 such as DMEM may be supplied to the fixed cells 117.

  Next, in S130, the fixed cells 117 are cultured. For example, the container 100 in which the cells 117 are fixed is held at a predetermined temperature (for example, 37 ° C.) for a predetermined time (for example, 24 hours), and incubation is performed.

  Next, in S140, the inside of the well 110 of the container 100 is washed. For example, the culture solution 118 in the well 110 is removed and washed with a washing solution such as PBS (−) at least once (for example, three times). Accordingly, the inside of the well 110 is washed before the cells 117 are detached from the medium 112, the separation work is easily performed, and impurities collected together with the cells are reduced. Note that S140 may be omitted.

  Next, in S150, at least a part of the solution in the well 110 is removed. For example, when the cleaning of S140 is executed, at least a part of the cleaning liquid remaining in the well 110 is removed. From the viewpoint of protecting the cells 117, at least a part of the washing solution may remain in the well 110. Alternatively, the culture medium 118 may be newly supplied after all the cleaning liquid in the well 110 is removed.

  When S140 is omitted and the culture medium 118 supplied in S120 remains in the well 110, a part of the culture medium 118 may be removed. By removing a part of the culture solution 118, the heat capacity of the contents of the well 110 is reduced, and the temperature of the medium 112 can be raised in a shorter time. Further, if the culture medium 118 is partially removed without being removed, the cells 117 in the well 110 can be protected.

  Next, in S <b> 160, the container 100 is heated by the heater 300. For example, the heat transfer plate 200 is mounted on the light transmission part 310 of the heater 300, and the container 100 is placed so that the positions of the plurality of through parts 220 of the heat transfer plate 200 correspond to the positions of the plurality of wells 110. Arranged above, the heater 300 generates heat. Thereby, the heat generated from the heater 300 is transmitted to the container 100 through the heat transfer plate 200.

  The heating of S160 may be continued until a predetermined temperature or heating time is achieved. For example, the process may proceed to S170 when the solution such as the medium 112 or the culture medium 118 in the container has reached a predetermined temperature (for example, 36 ° C.).

  Next, in S <b> 170, while heating the container 100 via the heat transfer plate 200, the cells 117 to be peeled in the well 110 are irradiated with light from the light irradiation unit 400, and the cells 117 are removed from the medium 112. Peel off. For example, while observing the inside of the well 110 with the observation unit 800 being magnified, the cells 117 to be peeled are specified, and laser light is emitted from the light irradiation unit 400 toward the specified cells 117 to be peeled.

  Light irradiation from the light irradiation unit 400 may be continued until a predetermined temperature or irradiation time is achieved. As an example, a laser beam having a wavelength of 532 nm is irradiated for 10 seconds at an intensity of 3 mW to the cells 117 to be peeled.

  The light irradiation unit 400 causes the emitted light to enter the heater 300 through the optical member 500, the mirror 600, and the lens 700, and is transmitted through the light transmission unit 310 of the heater 300 and the penetration unit 220 of the heat transfer plate 200. Then, the cells 117 to be peeled in the well 110 are irradiated with light. The medium 112 in the vicinity of the cell 117 irradiated with light is heated and fluidized by light irradiation from the light irradiation unit 400 in addition to heat from the heater 300. As a result, the light irradiation unit 400 peels the cells 117 to be peeled from the medium 112.

  Here, before / between light irradiation, the cell peeling apparatus 10 may adjust the position and focus of irradiation light. For example, the driving unit 350 may adjust the light irradiation position of the light irradiation unit 400 so as to coincide with the position of the cell 117 to be peeled by moving the heater 300 functioning as a stage in the main surface direction.

  Further, the position of light irradiation of the light irradiation unit 400 may be changed by operating the galvanometer mirror of the optical member 500. Moreover, the cell peeling apparatus 10 may adjust optical conditions, such as a focus of a laser beam, by adjusting the optical member 500, the lens 700, and / or other adjustment apparatuses.

  In S170, instead of performing light irradiation on one cell 117 to be peeled, light irradiation may be collectively performed on a plurality of cells 117 to be peeled. For example, the light irradiation pattern data corresponding to the cell 117 to be peeled is created in advance, the light irradiation pattern is realized by the DMD or SLM of the optical member 500, and the light irradiation unit 400 passes through the light irradiation pattern. The well 110 may be irradiated with light. Thereby, the light irradiation part 400 can peel all the cells 117 of the peeling object in the well 110 by one light irradiation.

  Next, in S180, it is determined whether or not the light irradiation to all the cells 117 to be detached has been completed. If completed, the process proceeds to S200, and if not, the process proceeds to S190.

  In S190, the light irradiation target by the light irradiation unit 400 is changed. For example, the driving unit 350 may change the light irradiation position of the light irradiation unit 400 to another undetached cell 117 to be peeled by moving the heater 300 in the main surface direction. Further, the optical member 500 may operate the galvanometer mirror to change the light irradiation position of the light irradiation unit 400 to another undetached cell 117 to be peeled.

  In S200, the cell acquisition unit 900 acquires the detached cell 117. For example, while confirming the position of the cell 117 peeled by the observation unit 800, the cell acquisition unit 900 approaches the peeled cell 117 at the tip of the capillary, sucks the cell 117 with the capillary, and then sucks the cell 117. The cells 117 may be obtained by discharging the cells.

  The cell acquisition unit 900 may acquire a plurality of detached cells 117 in one or a plurality of wells 110 at a time instead of collecting the cells 117 one by one. For example, all the detached cells 117 in the container 100 may be collectively collected in the collection container by moving the container 100 onto the collection container and covering the container 100.

  In this way, the cell detachment method according to the present embodiment is realized by executing the processes of S110 to S200 using the cell detachment apparatus 10. According to the cell detachment method of the present embodiment, the medium 112 is rapidly heated by light irradiation while the medium 112 that is fluidized by heating through the heat transfer plate 200 is gently heated, so that the medium 112 is fluidized. The time of light irradiation with high invasiveness to the cells 117 can be made shorter than before, and the target cells 117 can be obtained more reliably.

  FIG. 8 shows an example of the effect of the cell peeling method of the present embodiment. The horizontal axis of the graph in FIG. 8 indicates the time (seconds) from the start of heating by the heater 300, and the vertical axis indicates the measurement result (° C.) of the surface temperature of the medium 112. The measurement was performed under the condition that only the medium 112 was accommodated in the well 110 and the culture solution 118 was not filled.

  Moreover, from the light irradiation part 400, the laser beam of 532 nm was continuously irradiated with the intensity of 5 mW from the time of about 180 seconds. The dotted line graph in FIG. 8 shows the measurement result obtained by placing the container 100 directly on the heater 300 without the heat transfer plate 200, and the straight line graph shows the aluminum heat transfer plate 200 between the heater 300 and the container 100. The result of having been measured between them is shown.

  As shown in the graph, under the condition where there is no heat transfer plate 200, the temperature rise until the light irradiation is slow, and the fluidization temperature of the medium 112 reaches 40 ° C. even in the vicinity of 250 seconds when the light irradiation is continued for a while. Absent. Therefore, in the absence of the heat transfer plate 200, it is necessary to increase the light irradiation time and increase the light intensity, and the invasiveness to the cells 117 is increased.

  On the other hand, when the heat transfer plate 200 was used, the temperature was raised relatively quickly before the light irradiation, and reached 40 ° C. in about several tens of seconds after the light irradiation. When the heat transfer plate 200 is used in this way, the light irradiation time can be made relatively short.

  For example, according to the cell detachment method of this embodiment, the light irradiation intensity can be reduced to 1/12 and the light irradiation time can be reduced to 1/6 as compared with the method not using the heat transfer plate 200. I was able to. Moreover, it has confirmed with respect to the Hela cell and MDCK cell that the cell has manufactured even after the cell exfoliation by applying the cell exfoliation method of this embodiment.

  As mentioned above, although this invention was demonstrated using embodiment, the technical scope of this invention is not limited to the range as described in the said embodiment. It will be apparent to those skilled in the art that various modifications or improvements can be added to the above-described embodiment. It is apparent from the scope of the claims that the embodiments added with such changes or improvements can be included in the technical scope of the present invention.

  The order of execution of each process such as operations, procedures, steps, and stages in the apparatus, system, program, and method shown in the claims, the description, and the drawings is particularly “before” or “prior to”. It should be noted that the output can be realized in any order unless the output of the previous process is used in the subsequent process. Regarding the operation flow in the claims, the description, and the drawings, even if it is described using “first”, “next”, etc. for convenience, it means that it is essential to carry out in this order. It is not a thing.

  10 cell peeling device, 100 container, 110 well, 112 medium, 114 gel, 116 gold fine particle, 117 cell, 118 culture solution, 200 heat transfer plate, 210 main body part, 212 core layer, 214 surface layer, 216 surface layer, 220 Penetration part, 300 heater, 310 light transmission part, 350 drive part, 400 light irradiation part, 500 optical member, 600 mirror, 700 lens, 800 observation part, 900 cell acquisition part

Claims (12)

A heat transfer plate on which a container having a plurality of wells in which cells are stored is placed,
The thermal conductivity as the bulk of the material constituting the heat transfer plate is higher than the thermal conductivity as the bulk of the material constituting the container,
A main body that contacts at least a portion of the outer bottom surface of each of the plurality of wells;
A heat transfer plate having a plurality of transparent penetrating parts configured to penetrate the main body part below at least a part of an inner bottom surface of each of the plurality of wells.   The heat transfer plate according to claim 1, wherein each of the plurality of through portions is an opening provided so as to penetrate the main body portion.   The heat transfer plate according to claim 2, wherein the plurality of through portions are filled with a transparent material.   The heat transfer plate according to any one of claims 1 to 3, wherein the main body portion covers at least the bottom surfaces of the outer walls of the plurality of wells. A container having a plurality of wells;
A heat transfer plate on which the container is placed;
A heater disposed in contact with the heat transfer plate;
A light irradiator for irradiating a target region in the well;
With
The thermal conductivity as a bulk of the material used for the heat transfer plate is:
A body part that is higher in thermal conductivity as a bulk of the material used for the container and is in contact with at least a part of the outer bottom surface of each of the plurality of wells;
A cell detaching apparatus comprising: a plurality of transparent penetrating parts penetrating the main body part below at least a part of an inner bottom surface of each of the plurality of wells. A cell acquisition unit for acquiring cells;
The cell peeling apparatus according to claim 5. The heater has a light transmission part that transmits light from the light irradiation part,
The cell peeling apparatus according to claim 5 or 6. An observation unit for magnifying the inside of the well;
The cell peeling apparatus according to any one of claims 5 to 7. A container having a plurality of wells for storing cells;
A heat transfer plate on which the container is placed;
The heat transfer plate is
A body portion having a higher thermal conductivity than the container and contacting at least a part of an outer bottom surface of each of the plurality of wells;
A well plate unit having a plurality of transparent through portions penetrating through the main body portion below at least a part of the bottom surface inside each of the plurality of wells.   The well plate unit according to claim 9, wherein each of the plurality of through portions is an opening provided so as to penetrate the main body portion.   The well plate unit according to claim 10, wherein the opening is filled with a transparent material.   The well plate unit according to any one of claims 9 to 11, wherein the main body covers at least the bottom surfaces of the outer walls of the plurality of wells.

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