WO2014092127A1 - がん細胞単離装置及びがん細胞単離方法 - Google Patents
がん細胞単離装置及びがん細胞単離方法 Download PDFInfo
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- WO2014092127A1 WO2014092127A1 PCT/JP2013/083232 JP2013083232W WO2014092127A1 WO 2014092127 A1 WO2014092127 A1 WO 2014092127A1 JP 2013083232 W JP2013083232 W JP 2013083232W WO 2014092127 A1 WO2014092127 A1 WO 2014092127A1
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- filter
- cancer cell
- cancer cells
- cell isolation
- cancer
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/04—Cell isolation or sorting
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/4077—Concentrating samples by other techniques involving separation of suspended solids
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- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
- G06T7/00—Image analysis
- G06T7/0002—Inspection of images, e.g. flaw detection
- G06T7/0012—Biomedical image inspection
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- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
- G06T7/00—Image analysis
- G06T7/10—Segmentation; Edge detection
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- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06V—IMAGE OR VIDEO RECOGNITION OR UNDERSTANDING
- G06V20/00—Scenes; Scene-specific elements
- G06V20/60—Type of objects
- G06V20/69—Microscopic objects, e.g. biological cells or cellular parts
- G06V20/693—Acquisition
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/4077—Concentrating samples by other techniques involving separation of suspended solids
- G01N2001/4088—Concentrating samples by other techniques involving separation of suspended solids filtration
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- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
- G06T2207/00—Indexing scheme for image analysis or image enhancement
- G06T2207/30—Subject of image; Context of image processing
- G06T2207/30004—Biomedical image processing
Definitions
- the present invention relates to a cancer cell isolation device and a cancer cell isolation method.
- CTC circulating tumor cells
- CTC cancer cells
- the present invention has been made in view of the above, and provides a cancer cell isolation device and a cancer cell isolation method that can isolate cancer cells captured on a filter without damaging them. For the purpose.
- a cancer cell isolation device isolates cancer cells captured on a filter having a plurality of through holes and stores them in a storage container.
- An isolation device based on an image picked up by the image pick-up means, an image pick-up means for enlarging and picking up the filter that has captured the cancer cells, a moving means that can move the filter and the storage container, And handling means for carrying the cancer cells one by one while moving the filter and the storage container by the moving means.
- the cancer cell isolation method is a cancer cell isolation method for isolating cancer cells captured on a filter provided with a plurality of through holes and storing them in a storage container.
- the filter and the storage container are moved by the moving means based on the captured image. And moving cancer cells one by one using handling means. Thereby, it can isolate without damaging the cancer cell on a filter, and observation etc. can be performed suitably.
- the handling means includes a suction nozzle, and the cancer cell is adsorbed to the tip of the suction nozzle by adsorbing the cancer cells.
- a mode of transporting cancer cells is mentioned.
- the handling means includes a microtweezer, and the cancer cell is held by holding the cancer cell with the microtweezer.
- the aspect to convey is mentioned.
- the image processing apparatus includes position recognition means for acquiring position information for specifying a position of the cancer cell on the filter based on an image of the filter imaged by the imaging means, and the information acquired by the position recognition means
- the cancer cells on the filter can be transported by the handling means based on the position information of cancer cells.
- the cancer cell is transported using the position information. It can be recognized and transport work can be performed efficiently.
- the filter is provided with a reference mark indicating the arrangement of the through holes, and based on the reference mark, the position recognition unit acquires the position information of the cancer cells, and the moving unit sets the filter. It can also be set as the aspect which moves.
- a cancer cell isolation device and a cancer cell isolation method that can isolate cancer cells captured on a filter without damaging them.
- FIG. 1 shows the configuration of a cancer cell isolation device according to an embodiment of the present invention.
- the cancer cell isolation device 1 is a device that moves the cancer cells X on the filter 100 in which cancer cells are captured one by one to a plurality of storage units 201 provided in the storage container 200.
- the cancer cell isolation device 1 includes a moving table 11 for moving the filter 100, a moving table control unit 20 for controlling the operation of the moving table 11, a camera 30 for magnifying and observing the filter 100, and a camera 30. And an output unit 32 that outputs the image captured in step 1 and a handling unit 40 for moving the cancer cells X from the filter 100 to the storage container 200.
- the moving table 11 includes a rotary moving table 12 that rotates and moves the filter 100 along a horizontal plane, and an X linear moving table 14 and a Y linear moving that move the filter 100 and the storage container 200 in two directions perpendicular to each other along the horizontal plane.
- the table 16 is configured to include a Z linearly moving table 18 that moves the filter 100 and the storage container 200 in the vertical direction. These operate based on an instruction from the moving table control unit 20 to move the filter 100 and the storage container 200, and form a situation in which the handling unit 40 can easily operate the cancer cells X.
- the camera 30 is attached so as to magnify and image the main surface of the filter 100 from above.
- the image captured by the camera 30 is output from the output unit 32 connected to the camera 30 so that an operator who operates the cancer cell isolation device 1 can confirm the image. Yes.
- the handling unit 40 includes a handling nozzle 41 (suction nozzle), a negative pressure pump 43 connected to the handling nozzle 41 via a tube 42, and a nozzle moving base 44 for moving the handling nozzle 41.
- the A pressure gauge 45 is attached in the middle of the tube 42 and has a function of monitoring the gas pressure in the tube 42.
- a valve 46 is attached in the vicinity of the end of the tube 42 on the negative pressure pump 43 side.
- the tube 42 is bifurcated on the side of the handling nozzle 41 with respect to the valve 46.
- a valve 49, a regulator 50, and a compressed air tank 51 are attached to the tube 48 connected to the tube 42 in this order from the branch point side. That is, when the valve 46 is closed and the valve 49 is opened, the compressed air in the compressed air tank 51 is output while being regulated by the regulator 50, thereby allowing the compressed air to flow into the tube 42.
- the negative pressure pump 43 is driven with the valve 46 opened and the valve 49 closed, whereby the air in the tube 42 is drawn by the negative pressure pump 43.
- Cancer cells X can be adsorbed to the tip of the handling nozzle 41.
- the inside of the tube 42 becomes negative pressure, so by confirming the value of the pressure gauge 45, it is confirmed that the cancer cell X has adhered to the tip of the handling nozzle 41.
- the handling nozzle 41 in a state where the cancer cells X are adsorbed up to a place (for example, the storage unit 201 of the storage container 200) where the cancer cells X are to be placed by the linked operation of the nozzle movement base 44 and the mobile base 11.
- a place for example, the storage unit 201 of the storage container 200
- the valve 46 is closed and the valve 49 is opened.
- the compressed air is output from the compressed air tank 51 to fill the tube 42 with the compressed air.
- the cell X can be removed from the handling nozzle 41.
- the filter 100 is a plate-like member provided with a plurality of through holes in the thickness direction of the main surface, details of which will be described later.
- a filter after capturing cancer cells using the diameter of the through-holes by passing blood or the like as a test solution through the through-holes of the filter 100 using a filter unit to be described later, It is attached to the release device.
- the storage container 200 is a container for storing the isolated cancer cells X one by one and storing the cancer cells one by one in the partitioned storage unit 201. It is a container that can.
- the filter 100 is attached to the cancer cell isolation device 1. It is assumed that the cancer cell X has already adhered to the filter 100 at this time, and washing / staining treatment or the like is performed as necessary.
- the rotary moving table 12 is used. use.
- the moving directions of the X rectilinear moving table 14 and the Y rectilinear moving table 16 are orthogonal to each other.
- the through holes are arranged along straight lines orthogonal to each other in the filter 100 is described.
- the camera 30 is focused on the main surface of the filter 100, that is, the surface to which the cancer cells X are attached, and the position where the cancer cells X are attached to the main surface of the filter 100 is the camera.
- the camera 30 is focused on the cancer cell X by being moved by the X rectilinear moving table 14, the Y rectilinear moving table 16 and the Z rectilinear moving table 18 so as to be imaged.
- the magnification of the camera 30 at this time is preferably a magnification that allows focusing on both the main surface of the filter 100 and the cancer cells X.
- the tip of the handling nozzle 41 of the handling unit 40 is moved to the vicinity of the cancer cell X using the nozzle moving table 44.
- the distance between the tip of the handling nozzle 41 and the cancer cell X is about 10 ⁇ m.
- the tip of the handling nozzle 41 and the cancer cell X are brought into contact with each other by moving the filter 100 by the X rectilinear moving table 14, the Y rectilinear moving table 16, and the Z rectilinear moving table 18. Furthermore, by driving the negative pressure pump 43, the cancer cells X are adsorbed to the tip of the handling nozzle 41 by suction. Adsorption of the cancer cells X can be confirmed by the suction pressure measured by the pressure gauge 45.
- the X linear moving table 14 and the Y linear moving table 16 are used to lower the filter 100.
- the filter 100 arranged in the direction is moved, and instead the storage container 200 is moved so that the predetermined storage part 201 of the storage container 200 is disposed.
- the storage unit 201 and the cancer cell X at the tip of the handling nozzle 41 approach each other by moving the storage container 200 upward by the Z linearly moving table 18.
- the inside of the tube 42 is switched from negative pressure to pressurized by switching the opening and closing of the valves 46 and 49 and the supply of compressed air from the compressed air tank 51, and the compressed air moves from the tip of the handling nozzle 41 to the storage portion 201.
- the cancer cell X is moved. Thereby, one of the cancer cells X on the filter 100 is isolated and stored in the storage unit 201 of the storage container 200.
- the storage container 200 and the filter 100 are switched by driving the X linear movement table 14, the Y linear movement table 16, and the Z linear movement table 18, thereby moving the filter 100 below the camera 30 and performing the above operation.
- the isolation operation of the cancer cell X on the filter 100 is performed one by one.
- FIG. 2 is a diagram for explaining the configuration of the filter 100.
- the filter 100 shown in FIG. 2 has a plurality of substantially rectangular through holes 102 formed in the thickness direction of the main surface of the sheet 101.
- a plurality of through holes 102 are arranged in a matrix.
- alignment marks 103 used for alignment are arranged at the four corners of the sheet 101.
- individual identification characters 104 are printed on the sheet 101 in the vicinity thereof in association with each of the through holes 102.
- the alignment mark 103 indicates the arrangement of the through holes 102, that is, the direction of the sheet 101, and the movement direction of the X rectilinear movement table 14 and the Y rectilinear movement table 16. Is used when adjusting using the rotary moving table 12. That is, the alignment marks 103 arranged at the four corners of the filter 100 are used in association with the positional relationship (arrangement, rectangular shape of the through hole 102) of the filter 100. Specifically, for example, when the alignment mark 103 has a “cross” shape as shown in FIG. 2, the direction indicated by the cross is made to coincide with the short axis direction 102X and the long axis direction 102Y of the through hole 102. .
- the filter when the filter is fixed to the cancer cell isolation device 1 in a state where the cross alignment mark 103 is associated with the moving directions of the X rectilinear moving table 14 and the Y rectilinear moving table 16, the X rectilinear moving table 14 and Y Since the moving direction of the rectilinear moving table 16 coincides with the major axis direction and the minor axis direction of the through hole 102, it is easy to move the handling nozzle 41, move the filter 100, and the like more accurately.
- the identification character 104 written in the vicinity of the through hole 102 is a combination of two numbers in parentheses.
- the first number of the two numbers indicates the row of the through holes 102
- the second number indicates the column of the through holes 102.
- the through hole 102 on the surface of the filter 100 can be specified.
- the position of the through hole 102 is specified, so that the through hole 102 of the filter 100 in which the cancer cells X are captured using the identification character 104 of the through hole 102 is defined. It can be easily identified and the isolation operation can be performed efficiently. Note that the information for specifying the position of the through-hole 102 does not need to stick to the above-described two-numbered list.
- FIG. 3A and 3B are diagrams illustrating the configuration of the filter unit 300
- FIG. 3A is a view of the filter unit 300 as viewed from above
- FIG. 3B is a schematic cross-sectional view.
- FIG. 3 illustrates a filter unit 300 having a circular outer shape used when a cancer cell is captured using a filter 100 having a circular sheet outer shape.
- the filter unit 300 is configured by combining an upper nut 301 having a concave cross section and a cylindrical lower screw 302, and the filter 100 includes an upper surface 302a of the lower screw 302 and a bottom surface 301a of a concave portion of the upper nut 301. It is fixed so that it is pinched. On the bottom surface 301a and the top surface 302a, gaps 311 and 312 are formed at positions corresponding to the positions where the through holes 102 of the filter 100 are provided, respectively.
- the upper nut 301 is provided with an upper nut nozzle 321 that communicates with the gap 311 and supplies blood from the outside.
- the lower screw 302 is also provided with a lower screw nozzle 322 that communicates with the gap 312 and supplies blood from the outside.
- the test liquid such as blood is supplied from the end of the upper nut nozzle 321 after the filter 100 is sandwiched and fixed by the upper nut 301 and the lower screw 302.
- the test liquid that has reached the gap 311 from the upper nut nozzle 321 passes through the through hole 102 of the filter 100 and is discharged from the gap 312 to the outside via the lower screw nozzle 322.
- cancer cells having a diameter larger than that of the through hole 102 are captured by the through hole 102 of the filter 100.
- the cancer cells on the filter 100 are washed and stained by sequentially supplying treatment liquids for observing cancer cells such as a washing liquid and a staining liquid from the upper nut nozzle 321.
- the filter unit 300 can be taken out by separating the upper nut 301 and the lower screw 302 by performing the operation of loosening the screws of the upper nut 301 and the lower screw 302.
- the screw loosening operation may be performed by fixing the lower screw 302 to another fixing base (not shown) so as not to apply vibration to the filter 100.
- the cancer cell X captured on the filter 100 is imaged by the camera 30 serving as an imaging unit, and the captured image is output to the output unit. 32, while the filter 100 and the storage container 200 are moved by the X rectilinear moving table 14, the Y rectilinear moving table 16, and the Z rectilinear moving table 18, which are moving means, cancer cells using the handling unit 40 are output. Carries one X at a time. Thereby, it can isolate without damaging the cancer cell on a filter, and observation etc. can be performed suitably.
- FIG. 4 is a diagram for explaining a first modification of the cancer cell isolation device.
- the cancer cell isolation device 2 shown in FIG. 4 is different from the cancer cell isolation device 1 shown in FIG. 1 in the following points. That is, in place of the handling unit 40, in the cancer cell isolation device 2, the micro tweezers 55 is used as the handling means.
- the micro tweezers 55 is attached to a tweezer moving table 56, and the tip of the micro tweezers 55 is movable in the imaging region of the camera 30.
- the micro tweezers 55 is used as the handling means, as in the case where the handling nozzle 41 is used, after the camera 30 is focused on the cancer cells X on the main surface of the filter 100, In addition, after the tip of the microtweezers 55 is moved to the vicinity of the cancer cell X, the cancer cell X is gripped by the tip of the microtweezers 55, and then the filter 100 and the storage container 200 are moved, thereby causing cancer. The cancer cells X can be transported to the storage part 201 of the storage container 200.
- the cancer cell isolation device 3 shown in FIG. 5 is different from the cancer cell isolation device 1 shown in FIG. 1 in the following points. That is, information indicating the position of the cancer cell X on the filter 100 is acquired using an image captured by the camera 30, and based on this information, the moving table controller 20 rotates the moving table 12, the X linear moving table 14, The difference is that the Y linear moving table 16 and the Z linear moving table 18 are moved.
- an image captured by the camera 30 is sent to a recognition unit 62 (position recognition unit) connected to the output unit 32. Then, in the recognition unit 62, the through-hole 102 on which the cancer cell X is captured is specified.
- position information that specifies the position of the cancer cell X is acquired by the recognition unit 62.
- the positional information used here for example, the identification character 104 or the like given to each of the through holes 102 of the filter 100 can be cited, and the identification character 104 for identifying the through hole 102 where the cancer cell X is captured is used.
- Such information is acquired in the recognition unit 62 as position information of the cancer cell X.
- the positional information of the cancer cell X is sent from the recognition unit 62 to the control unit 60.
- an instruction related to the movement of the filter 100 and the storage container 200 for transporting the cancer cell X is performed. Specifically, an instruction relating to the movement of the moving base 11 for moving cancer cells X at a specific position of the filter 100 specified by the recognition unit 62 to the storage unit 201 of the storage container 200 is given.
- the control unit 60 may instruct the handling unit 40 to drive the handling nozzle 41.
- the moving table control unit 20 is driven according to an instruction from the control unit 60 and the handling nozzle 41 is operated, whereby the cancer cells X on the filter 100 can be moved from the filter 100 to the storage container 200.
- the acquisition of position information related to the position of the cancer cell X on the filter 100 and the isolation operation of the cancer cell are performed on the cancer cell isolation device 3.
- it was performed with the filter 100 fixed it is also possible to remove the filter 100 once after acquiring the position information related to the position of the cancer cell X on the filter 100.
- the cancer cell isolation device 3 holds the positional information of the cancer cells X on the filter 100.
- the isolation operation can be quickly performed by accurately attaching the filter 100 to the apparatus.
- the alignment mark 103 (see FIG. 2) provided on the filter 100 is useful.
- the filter 100A shown in FIG. 6 differs from the filter 100 shown in FIG. 2 in the following points. That is, straight lines 105X and 105Y extending in two directions orthogonal to each other are added so as to connect adjacent through holes 102.
- the straight line 105 ⁇ / b> X is a straight line that extends in the short side direction of the through hole 102 and connects the midpoints of the adjacent through holes 102.
- the straight line 105 ⁇ / b> Y is a straight line that extends in the long side direction of the through hole 102 and connects the midpoints of the adjacent through holes 102.
- the straight lines 105X and 105Y connecting the through holes 102 are described in the sheet 101, so that not only the alignment marks 103 provided at the four corners of the sheet 101 but also the straight lines 105X and 105Y are referred to.
- the filter 100 can be more accurately aligned, and the filter according to the position of the through hole 102 where the cancer cell X is captured. 100 movements can be performed more suitably.
- the shape of the through hole 106 on the surface of the sheet 101 is a wave shape.
- the corrugated through-hole is formed by connecting a plurality of perforations having a rectangular or rounded rectangular shape on the surface of the sheet 101 at a predetermined crossing angle between the ends.
- the corrugated shape of the through hole 106 is formed along the X-axis direction.
- FIG. 8 is a cross-sectional view showing a modification of the filter unit, and corresponds to FIG.
- the filter unit 300A shown in FIG. 8 differs from the filter unit 300 of FIG. 3 in the following points.
- the filter 100 is fixed between the upper nut 301 and the lower screw 302 in a state where the upper and lower edges of the filter 100 are sandwiched between the upper frame plate 331 and the lower frame plate 332.
- the upper frame plate 331 and the lower frame plate 332 are joined to the main surface (upper surface and lower surface) of the filter 100 so as to sandwich the filter 100.
- the filter 100, the upper frame plate 331, and the lower frame plate 332 are integrated.
- the upper frame plate 331 and the lower frame plate 332 are preferably members that do not cause a gap when the filter 100 is attached between the upper nut 301 and the lower screw 302.
- silicon rubber or the like can be used. .
- the upper frame plate 331 and the lower frame plate 332 fix the filter 100 in a state in which the filter 100 does not sag, it is possible to prevent the filter 100 from being deformed. Further, when the filter 100 is taken out from the filter unit 300A, it is taken out with the upper frame plate 331 and the lower frame plate 332 attached. For this reason, even when the filter 100 alone is moved, wrinkles can be prevented from being generated on the filter surface, and damage to the cancer cells X captured on the filter 100 can be prevented. When attaching to the cancer cell isolation device and also in the subsequent operation, it is possible to prevent a decrease in workability due to deformation of the filter 100.
- the filter unit 300A shown in FIG. 8 differs from the filter unit 300 of FIG. 3 in the following points. That is, the difference is that the filter 100 is fixed between the upper nut 301 and the lower screw 302 in a state where the filter 100 is sandwiched between the upper plate 341 and the lower plate 342 forming the gaps 311 and 312. An upper nut nozzle 321 connected to the gap 311 is attached to the upper plate 341, and a lower screw nozzle 322 connected to the gap 312 is attached to the lower plate 342. In such a configuration, the upper nut 301 and the lower screw 302 can be used a plurality of times because contact with the test solution is suppressed.
- a highly durable metal material can be used for the upper nut 301 and the lower screw 302.
- the upper plate 341 and the lower plate 342, which are considered to be in contact with the test solution are manufactured and used as, for example, resin consumables, and compared with the case where the filter unit 300 of FIG. 3 is used.
- the cost associated with cell isolation can also be reduced.
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Abstract
Description
図1に本発明の実施形態に係るがん細胞単離装置の構成を示す。がん細胞単離装置1は、がん細胞が捕捉されたフィルタ100上のがん細胞Xを1個ずつ収納容器200に設けられた複数の収納部201に移動させる装置である。がん細胞単離装置1は、フィルタ100を移動させる移動台11と、移動台11の動作を制御する移動台制御部20と、フィルタ100を拡大して観察するためのカメラ30と、カメラ30で撮像された画像を出力する出力部32と、がん細胞Xをフィルタ100から収納容器200へ移動させるためのハンドリング部40と、を含んで構成される。
ここで、本実施形態に係るがん細胞単離装置1を用いたがん細胞単離方法を説明する。まず、がん細胞単離装置1にフィルタ100を取り付ける。このときのフィルタ100には、既にがん細胞Xが付着しているものとし、必要に応じて、洗浄・染色処理等が行われているものとする。フィルタ100をがん細胞単離装置1に取り付ける際には、X直進移動台14及びY直進移動台16の移動方向とフィルタの貫通孔の並び方向とを合致させるために、回転移動台12を使用する。なお、X直進移動台14及びY直進移動台16の移動方向は互いに直交する方向となっている。なお、ここでは、フィルタ100において互いに直交する直線に沿って貫通孔が並んでいる場合について説明している。
ここで、図2を用いて、がん細胞の単離に用いられるフィルタ100について説明する。図2は、フィルタ100の構成を説明する図である。図2に示すフィルタ100は、シート101主面の厚さ方向に複数の略矩形の貫通孔102が形成されたものである。図2のフィルタ100では、貫通孔102が複数個マトリックス状に配置されている。また、位置合わせに用いられるアライメントマーク103がシート101の四隅に配置されている。また、貫通孔102それぞれに対応付けて、その近傍のシート101上に個別の識別文字104(基準マーク)が印字されている。
次に、図3を参照しながら上記のフィルタ100により血管中のがん細胞を捕捉するために用いられるフィルタユニットについて説明する。図3は、フィルタユニット300の構成を説明する図であり、図3(A)はフィルタユニット300を上方から見た図であり、図3(B)は概略断面図である。図3では、シートの外形が円形のフィルタ100を用いてがん細胞を捕捉する場合に用いられる外形が円形状のフィルタユニット300について説明する。フィルタユニット300は、断面が凹型形状の上ナット301と、円柱形状の下ネジ302とを組み合わせることで構成され、フィルタ100は、下ネジ302の上面302aと上ナット301の凹部の底面301aとに挟まれるようにして固定される。底面301aと上面302aとには、それぞれフィルタ100の貫通孔102が設けられる位置に対応する位置に空隙311,312が形成されている。また、上ナット301には、空隙311に対して連通して外方から血液を供給するための上ナットノズル321が設けられている。同様に、下ネジ302においても、空隙312に対して連通して外方から血液を供給するための下ネジノズル322が設けられている。
以下、上記のがん細胞単離装置及びがん細胞単離装置に用いられるフィルタに係る変形例を説明する。
図4は、がん細胞単離装置の第1の変形例を説明する図である。図4に示すがん細胞単離装置2は、図1に示すがん細胞単離装置1と比較して、以下の点が相違する。すなわち、ハンドリング部40に代えて、がん細胞単離装置2では、ハンドリング手段としてマイクロピンセット55が用いられている。マイクロピンセット55は、ピンセット移動台56に取り付けられ、その先端がカメラ30の撮像領域を移動可能とされている。このように、ハンドリング手段としてマイクロピンセット55を用いた場合でも、ハンドリングノズル41を用いた場合と同様に、フィルタ100の主面のがん細胞Xに対してカメラ30の焦点を合わせた後に、次に、マイクロピンセット55の先端をがん細胞Xの近傍まで移動させた後に、マイクロピンセット55の先端にてがん細胞Xを把持し、その後フィルタ100及び収納容器200を移動させることとでがん細胞Xを収納容器200の収納部201に対してがん細胞Xを運搬することができる。
次に、フィルタの変形例について、図6を用いて説明する。図6に示すフィルタ100Aは、図2に示すフィルタ100と比較すると以下の点が相違する。すなわち、隣接する貫通孔102を結ぶように、互いに直交する2方向に伸びる直線105Xと105Yとを追加した点である。直線105Xは、貫通孔102の短辺方向に伸びて隣接する貫通孔102の中点同士を結ぶような直線である。また、直線105Yは、貫通孔102の長辺方向に伸びて隣接する貫通孔102の中点同士を結ぶような直線である。このように、フィルタ100Aでは、貫通孔102を結ぶ直線105X,105Yをシート101に記載することで、シート101の四隅に設けられたアライメントマーク103だけでなく直線105X,105Yを参照しながら、X直進移動台14、Y直進移動台16との位置合わせを行うことで、フィルタ100をより正確に位置合わせすることができると共に、がん細胞Xが捕捉された貫通孔102の位置に応じたフィルタ100の移動をより好適に行うことができる。
次に、フィルタユニットの変形例について、図8を用いて説明する。図8はフィルタユニットの変形例を示す断面図であり図3(B)に対応する図である。図8に示すフィルタユニット300Aでは、図3のフィルタユニット300と比較して、以下の点が相違する。すなわち、フィルタ100の周縁部の上下が上枠板331と下枠板332とで挟まれた状態で、上ナット301と下ネジ302との間に固定されている点が相違する。図8のフィルタユニット300Aにおいて、上枠板331と下枠板332とは、フィルタ100を挟むように、フィルタ100の主面(上面と下面と)に対してそれぞれ接合されている。すなわち、フィルタ100、上枠板331及び下枠板332は、一体化されている。上枠板331及び下枠板332は、上ナット301と下ネジ302との間にフィルタ100を取り付けた際に隙間が生じない部材であることが好ましく、例えば、シリコンゴム等を用いることができる。
Claims (6)
- 貫通孔が複数設けられたフィルタ上において捕捉されたがん細胞を単離して収納容器に収納するがん細胞単離装置であって、
前記がん細胞を捕捉したフィルタを拡大して撮像する撮像手段と、
前記フィルタ及び前記収納容器を移動可能な移動手段と、
前記撮像手段により撮像された画像に基づいて、前記移動手段により前記フィルタ及び前記収納容器を移動させながら前記がん細胞を1個ずつ運搬するハンドリング手段と、
を備えることを特徴とするがん細胞単離装置。 - 前記ハンドリング手段は、吸引ノズルを含んで構成され、
前記吸引ノズルの先端に対して前記がん細胞を吸着させることで前記がん細胞を運搬する
ことを特徴とする請求項1記載のがん細胞単離装置。 - 前記ハンドリング手段は、マイクロピンセットを含んで構成され、
前記マイクロピンセットにより前記がん細胞を把持することで前記がん細胞を運搬する
ことを特徴とする請求項1記載のがん細胞単離装置。 - 前記撮像手段により撮像された前記フィルタの画像に基づいて前記フィルタ上の前記がん細胞の位置を特定する位置情報を取得する位置認識手段を備え、
前記位置認識手段により取得された前記がん細胞の位置情報に基づいて前記ハンドリング手段により前記フィルタ上の前記がん細胞を運搬する
ことを特徴とする請求項1~3のいずれか一項に記載のがん細胞単離装置。 - 前記フィルタには前記貫通孔の配列を示す基準マークが設けられ、
前記基準マークに基づいて、前記位置認識手段によって前記がん細胞の位置情報を取得すると共に、前記移動手段によって前記フィルタを移動させる
ことを特徴とする請求項4記載のがん細胞単離装置。 - 貫通孔が複数設けられたフィルタ上において捕捉されたがん細胞を単離して収納容器に収納するがん細胞単離方法であって、
前記がん細胞を捕捉したフィルタを拡大して撮像する撮像ステップと、
前記撮像ステップにおいて撮像された画像に基づいて、前記フィルタ及び前記収納容器を移動させながら前記がん細胞を1個ずつ運搬するステップと、
を含むことを特徴とするがん細胞単離方法。
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