JP4956289B2 - Fine particle screening apparatus and fine particle screening method - Google Patents

Description

  The present invention relates to a screening apparatus for fine particles, and in particular, applies light to fine particles (for example, cells) to search for fine particles targeted by using fluorescence emitted from the fine particles as a label. The present invention relates to a fine particle screening apparatus and a fine particle screening method for selectively sucking and collecting.

Search and collection devices that search and collect cells that are target objects include the following. A sample plate on which cells are placed is placed on the sample stage, and the illumination and the objective lens are arranged at positions facing each other across the sample plate. The nozzle for sucking the cells is arranged obliquely downward with respect to the cells so as to be sucked. The sample stage can be moved by an XY stage. In the search and recovery device, one sample plate is placed on the sample stage and illuminated, and the fluorescence of the cells is observed, and the cells that emit fluorescence are collected using a nozzle (see, for example, Patent Document 1).
JP-A-2005-207986

By the way, it is desired that the member on which the sample is placed is accurately positioned with respect to the optical system including the illumination and the objective lens, thereby improving the positioning workability and preventing the defocusing and the deviation of the recognition position.
For example, when measuring with a camera or the like, it is necessary to widen the area that can be measured at a time for high throughput. In order to accurately position this region within the field of view and perform measurement with the observation focus being adjusted over the entire surface, it is necessary to satisfy both the accuracy of the member itself and the positioning accuracy during setting.

  Therefore, in order to solve the above-mentioned problems, the present invention improves positioning workability by preventing positioning from being out of focus and recognizing position by positioning with high accuracy, and can be targeted from a large number of fine particles. It is an object of the present invention to provide a fine particle screening device and a fine particle screening method capable of efficiently searching and selectively recovering particles.

In order to solve the above-described problems, the fine particle screening apparatus of the present invention searches for fine particles using light information emitted from the fine particles as a label, and selects fine particles for selectively acquiring the searched fine particles. A screening device,
A measuring chip provided with a plurality of wells formed of a material that transmits light and having a size capable of accommodating at least one of the fine particles;
A fixture for fixing the measurement chip;
A measurement unit that obtains an optical signal of the fine particles by irradiating light to the fine particles accommodated in the measurement chip and the measurement chip; and
The measuring chip is
A protrusion provided on a side surface of the measurement chip in order to be positioned in a direction parallel to the upper and lower surfaces of the measurement chip by making contact with the abutting portion of the fixture;
And a display unit which is formed on the measurement chip and clearly indicates a direction when the measurement chip is set on the fixture.

The fine particle screening apparatus of the present invention is preferably characterized in that the protrusion is formed so as to protrude from a side surface of the measuring chip.
In the fine particle screening apparatus of the present invention, preferably, the measurement chip has an area in which the well is formed having a width of 10 mm square or more, and a part or all of the area around the well is fixed. The flatness of an arbitrary 2 mm square in the area where the well is formed is 10 μm or less by applying a force of an appropriate size from the lower surface side where the well is not formed. And

The fine particle screening apparatus of the present invention preferably has a thickness variation Δt in an arbitrary area of 2 mm square included in the measurement area where the well of the measurement chip is formed, at 10 / (1-1 / n) [μm] or less (n is the refractive index of the material ratio of the measuring chip).
The fine particle screening apparatus of the present invention preferably has a variation in the inclination angle φ formed by the upper surface and the lower surface in an arbitrary 2 mm square region included in the measurement area in which the well of the measurement chip is formed. , (5 × 10 ⁻⁶ ) / {t (1-1 / n)} [rad] or less (t is the average thickness [mm] of the measurement chip, and n is the refractive index of the material of the measurement chip. ).

  The fine particle screening method of the present invention is a fine particle screening method for searching for fine particles using light information emitted from the fine particles as a label, and selectively acquiring the searched fine particles, for measurement. The chip is formed of a material that transmits light, and a plurality of wells having a size capable of storing at least one of the fine particles are provided. The measurement chip is fixed to a fixture, and the measurement unit However, when the optical signal of the fine particle is obtained by irradiating the fine particle accommodated in the measurement chip and the measurement chip with the light, the protrusion of the measurement chip is connected to the fixing tool. When positioning by abutting against the abutment portion, the positioning device is set with respect to the fixing device based on a display unit that clearly indicates a setting direction with respect to the fixing device. That.

  According to the present invention, by positioning accurately, the positioning workability is improved, the defocusing and the recognition position are prevented from being shifted, and the target fine particles are efficiently searched from a large number of fine particles. It can be selectively and efficiently collected.

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the drawings.
FIG. 1 is a front view showing a preferred embodiment of the fine particle screening apparatus of the present invention. FIG. 2 is a perspective view showing a preferred embodiment of the fine particle screening apparatus of FIG.

  The fine particle screening apparatus 1 shown in FIG. 1 and FIG. 2 uses a fluorescent light emitted from a fine particle as a label by applying light to a plurality of fine particles (for example, living cells) in the measurement chip 90 as a target. This is a device for searching for fine particles and selectively aspirating all fine particles in a well containing fine particles satisfying the collection conditions as a target object and collecting them on a collection plate 80. .

  The fine particle screening apparatus 1 shown in FIG. 1 includes a base 11, a support part 12, a recovery part 13, a measurement part 14, an image analysis part 15, and a moving part 16, as shown in FIG. In addition, each part is covered with a cover 19. The cover 19 prevents light and foreign matter from entering from the outside. The base 11 is a main body frame for holding each element of the screening apparatus 1.

  As shown in FIG. 1, the vertical direction in FIG. 1 is the X direction (first direction), and the left-right direction is the Y direction (second direction). The Z direction is a direction perpendicular to the X direction and the Y direction.

  The base 11 holds the collecting unit 13, the measuring unit 14, and the moving unit 16, and the base 11 has plate members 20, 21, and 22. These plate members 20 and 21 are fixed in parallel by a plurality of vertical members 23, and the plate members 20 and 22 are fixed in parallel by a plurality of members 24. The member 24 is made of a material that adjusts the height and blocks vibration.

  On the uppermost plate member 21, the support portion 12 and the support base 30 are fixed. The support part 12 is vertically arranged on the plate member 21 along the Z direction. The support base 30 has a leg portion 30R and a support plate 30P. The plate members 20, 21, 22 and the support plate 30P are arranged at intervals from each other along the Z direction.

Next, a structural example of the moving unit 16 will be described with reference to FIGS. 1 and 3.
FIG. 3 shows the moving unit 16, the mounting table 4 mounted on the moving unit 16, the recovery plate 80, and the measuring chip 90.

  3 and 1 is fixed to a support 30 of the base 11 shown in FIG. The moving unit 16 is equipped with a mounting table 40 that is a moving object, and the mounting table 40 can be moved and positioned along the X direction and the Y direction, respectively.

  As shown in FIG. 3, the moving unit 16 includes a first table 51 and a second table 52. The first table 51 is fixed to the support base 30 in FIG. 1, and the second table 52 is mounted so as to be positioned by moving along the X direction. The second table 52 is mounted so that the mounting table 4 can be positioned by moving along the Y direction.

  As shown in FIG. 3, guide rails 53 and 53 and a first motor 55 are provided on the upper surface of the first table 51. On the lower surface of the second table 52, members 54 and 54 having a U-shaped section and a nut 57 are provided. Guide rails 53 and 53 are engaged with members 54 and 54, respectively. The feed screw 56 of the first motor 55 is engaged with the nut 57. Thereby, the control part 100 commands and operates the first motor 55 to rotate the feed screw 56, whereby the second table 52 can be moved and positioned along the X direction.

  As shown in FIG. 3, guide rails 63 and 63 and a second motor 65 are provided on the upper surface of the second table 52. On the lower surface of the mounting table 40, U-shaped members 64, 64 and nuts 67 are provided. The guide rails 63 and 63 are engaged with the members 64 and 64, respectively. The feed screw 66 of the second motor 65 is engaged with the nut 67. Thus, the mounting table 40 can be moved and positioned along the Y direction by the control unit 100 instructing to operate the second motor 65 and rotate the feed screw 66.

  As shown in FIG. 3, the first table 51 has a first opening 59, the second table 52 has a second opening 69, and the mounting table 40 has a third opening 79. have. The first opening 59, the second opening 69, and the third opening 79 are sized so that they always overlap even when the second table 52 moves in the X direction and the mounting table 40 moves in the Y direction. It is an opening for guiding light (irradiation light) L from the objective lens 110 side of the measurement unit 14 to fine particles of the measurement chip 90 on the mounting table 40.

  As a result, even if the second table 52 moves in the X direction and the mounting table 40 moves in the Y direction, the light L from the objective lens 110 side causes the first opening 59, the second opening 69, and the third. The fine particles of the measuring chip 90 on the mounting table 40 are irradiated through the opening 79, and fluorescence can be generated from the fine particles.

  The mounting table 40 of the moving unit 16 shown in FIG. 3 can be moved in the X and Y directions using a motor and a feed screw, but is not limited thereto, and a linear motor or the like may be used.

  FIG. 4 shows the mounting table 40, the recovery plate 80, and the measuring chip 90. As shown in FIG. 4, the mounting table 40 is, for example, a rectangular plate-like member. On the mounting surface 70 of the mounting table 40, a collection plate 80 and a measuring chip 90 are detachable in the Y direction. Can be mounted side by side.

  The collection plate 80 is a plate-like member, and a large number of wells 81 are arranged on the collection plate 80 in a matrix form at equal intervals along the X direction and the Y direction. These wells 81 can collect and store fine particles, such as biological cells, which are sequentially discharged and discharged separately from the suction / discharge capillary 140. It is a storage unit. The well 81 is, for example, a substantially U-shaped recess or a cup-shaped recess when viewed in cross section.

  The measurement chip 90 shown in FIG. 4 is fixed by a fixing tool 120, and the fixing tool 120 is positioned and fixed at a predetermined position of the mounting table 40.

  FIG. 5 is a cross-sectional view showing an example of the shape of the measuring chip 90 and the fixture 120 of the measuring chip 90.

  The fixture 120 is for fixing and holding the measuring chip 90 at the position of the reference surface CL having a certain height with respect to the mounting surface 70 of the mounting table 40. In the example of FIG. 5, the fixing member 120 includes a first case part 121, a second case part 122, and an elastic member 123. The elastic member 123 is, for example, a rectangular ring, and the elastic member 123 is fixed to the flat surface 126 inside the second case portion 122.

  The measurement chip 90 is disposed between the first case part 121 and the second case part 122. When the measurement chip 90 is pushed up in the Z1 direction by the elastic member 123, the upper surface 90S of the measurement chip 90 is The first case portion 121 is abutted against the flat inner surface 124.

  As a result, the upper surface 90S of the measurement chip 90 is always pressed against the reference surface 124 with high positional accuracy, and the upper surface 90S of the measurement chip 90 can be positioned on the reference surface CL. Even if there are variations in thickness and warpage, the effects of thickness variation and warpage can be reduced.

  Therefore, the distance in the Z direction between the upper surface 90S of the measuring chip 90 and the objective lens 110 and the collection plate 80 of the measuring unit 14 can be accurately managed. In other words, the distance between the position of the fine particles M in the well 200 of the measurement chip 90 shown in FIG. 7 and the objective lens 110 and the recovery plate 80 of the measurement unit 14 can be accurately managed.

  The first case portion 121 can be opened and closed with respect to the second case portion 122 using, for example, a hinge mechanism portion, whereby the measurement chip 90 in the fixture 120 is taken out and a new measurement chip is obtained. The structure can be exchanged for 90.

Next, the measurement chip 90 will be described with reference to FIGS. 3, 5, and 7.
An example of the shape of the measurement chip 90 is illustrated in FIG. 7A, and the measurement chip 90 is made of a light-transmitting material such as glass or plastic.

  The measurement chip 90 shown in FIGS. 3 and 5 is made of a material that transmits light, and is, for example, a rectangular plate-like member. A plurality of wells 200 are arranged in the X direction and the Y direction to store a plurality of fine particles, respectively. They are arranged in a matrix at regular intervals along the direction. As shown in FIG. 5, each well 200 has such a size that at least one of the fine particles M can be stored by dispensing or batch-injecting a suspension 500 of fine particles.

  It is desirable that a plurality of recesses shallower than the depth of the well 200 be formed on the upper surface 90S of the measurement chip 90. The shallow concave portions are used as a reference for the measurement unit 14 to autofocus the position of the measurement chip 90 in the Z direction. That is, a recess as an autofocus marking is formed in a portion other than the well on the upper surface of the measurement chip 90, but the depth of the recess is preferably as steep and shallow as possible when performing contrast analysis by image processing. For example, 3 μm or less is preferable. The concave portion may be formed at a fixed position on the upper surface of the measurement chip 90 or may be a random scratch. The formation position of the concave portion should be determined in consideration of the adsorptivity of the measurement target and the like. It is.

  As shown in FIGS. 1 and 5, the measuring chip 90 is fixed to the moving unit 16 using a fixing tool 120. In this fixture 120, at least a surface (hereinafter referred to as a reference surface) 124 that is in contact with the surface on which the well 200 of the measurement chip is formed is formed of a material having higher rigidity than the measurement chip 90. The measurement chip 90 abuts the upper surface 90S on which the well 200 is formed on the reference surface CL of the fixture 120 and presses it with the elastic member 123 having elasticity from the surface opposite to the side on which the well 200 is formed. Further, the seal member 222 seals between the reference surface CL of the fixture 120 and the upper surface 90S of the measuring chip 90. With this fixing structure, the warping of the measuring chip 90 itself is corrected, the positioning with respect to the fixing tool 90 is performed, and furthermore, the sealing 500 is surely sealed so as not to drop the suspension 500 dripping from the upper surface 90S of the measuring chip 90 Can do.

  As illustrated in FIG. 7A, each well 200 is a fine particle collection and storage unit for storing fine particles M such as biological cells. For example, the cross-sectional shape is a substantially U-shaped recess or cup. It is a recess of the mold. Each well 200 stores a plurality of fine particles M, and a film of the suspension 500 is formed on the upper surface 90S of the measuring chip 90. The diameter D of each well 200 is, for example, 10 μm, and 500 wells are arranged in the X direction and the Y direction, respectively.

Next, with reference to FIG. 1 and FIG. 6, the structural example of the collection | recovery part 13 is demonstrated.
The recovery unit 13 shown in FIG. 1 includes a suction / discharge capillary 140 that is disposed on the base 11 and can be positioned by moving in the Z direction orthogonal to the X direction and the Y direction. Thus, the total number of fine particles in the well 200 in which the fine particles M1 satisfying the luminance condition illustrated in FIG. 5 are stored is sucked and discharged to a predetermined position of the collection plate 80 and collected.

  The collection unit 13 has a suction / discharge capillary 140 that can be positioned by moving in the Z direction orthogonal to the X direction and the Y direction. The fine particles satisfying the luminance condition by the suction / discharge capillary 140. The total number M of fine particles in the well in which M1 is stored is sucked and discharged to a predetermined position of the recovery plate 80 for recovery. That is, data for confirming that at least one particle satisfying the luminance condition that can be set by the measurer exists in at least each well, and the collection unit 13 collects all the fine particles in the well. Is.

  As shown in FIG. 6, the collection unit 13 includes an operation unit 130 and a base 131, and the base 131 is fixed to the support unit 12. The operation unit 130 includes a suction pump 133 and a suction / discharge capillary 140. The suction / discharge capillary 140 is a hollow member tapered in the Z2 direction (downward), and the tip 142 of the suction / discharge capillary 140 is a well of the measurement chip 90 as shown in FIGS. 200, and the rear end 143 of the suction / discharge capillary 140 is connected to the suction pump 133.

  As shown in FIG. 6, the feed screw 252 of the motor 251 is engaged with the nut 253 of the operation unit 130, and the control unit 100 operates the motor 251 to rotate the feed screw 252. Can be positioned by moving up and down along the Z direction (Z1 direction and Z2 direction).

  Accordingly, the collection unit 13 in FIG. 6 can be called a Z stage, and the moving unit 16 in FIG. 3 can also be called an XY stage.

Next, the measurement unit 14 and the analysis unit 15 will be described with reference to FIG.
1 irradiates light L to a region including a plurality of wells 200 of the measurement chip 90, thereby generating fluorescence from the fine particles M in the region, and emitting the fluorescence. Receive light. The fluorescence from the received fine particles M is image-analyzed by the image analysis unit 15. The image analysis unit 15 calculates the fluorescence intensity of the fine particles M1 that emit fluorescence of at least the maximum luminance among the plurality of fine particles M in each well 200.

  As a result, the control unit 100 recognizes the position of the well 200 in which the fine particles M1 that emit fluorescence with the maximum luminance satisfying the collection condition are stored in the plane constituted by the X direction and the Y direction. The control unit 100 gives control drive signals to the first motor 55 and the second motor 65 shown in FIG. 3 so that the well 200 of the measuring chip 90 on the moving unit 16 is directly below the suction / discharge capillary 140. Can be located. That is, the suction / discharge capillary 140 can suck fine particles in the well in units of wells. The suction / discharge capillary 140 sucks all the fine particles in the well containing the fine particles satisfying the collection condition. The suction / discharge capillary 140 can suck all fine particles from a selected well of the plurality of wells, that is, a well containing fine particles satisfying the collection condition.

  The measurement unit 14 shown in FIG. 1 irradiates the measurement chip 90 and the fine particles M accommodated in the measurement chip 90 with light guided from at least one light source, thereby allowing the shape of transmitted light or reflected light to be The position information and the luminance information such as fluorescence and chemiluminescence are acquired with a resolution finer than the average size of the individual fine particles, and the shape of the measurement chip itself and the position and size of the well 200 arranged on the measurement chip 90 are obtained. Get information such as.

  The image analysis unit 15 shown in FIG. 1 analyzes the measured shape information and optical information, so that at least one fine particle M1 satisfying a luminance condition that can be set to an arbitrary condition by the measurer is present in each well 200. Data for confirming that it exists is acquired. The fine particle screening apparatus 1 specifies the light information from the fine particles by matching the position recognition information of the well 200 with the transmitted light or reflected light and the light information of the fluorescence / chemiluminescence, and further, the measurement unit 14 has an autofocus function, performs measurement in a focused state, and determines the positional relationship between the tip 142 of the suction / discharge capillary 140 and the upper surface of the measurement chip 90 by performing autofocus on both. Can do.

  As shown in FIG. 1, the measurement unit 14 includes an objective lens 110, and the objective lens 110 guides light to the measurement chip 90. The objective lens 110 is disposed below the measurement chip 90 and the moving unit 16, and the suction / discharge capillary 140 is disposed above the measurement chip 90 and the moving unit 16. As a result, the measuring chip 90 and its moving part 16 can be disposed between the objective lens 110 and the suction / discharge capillary 140.

  In the measurement unit 14 of FIG. 1, for example, a laser light source or a mercury lamp can be adopted as the excitation light source 260. The shutter unit 261 is disposed between the excitation light source 260 and the fluorescence filter unit 262. When the shutter unit 261 does not irradiate the light L to the fine particles M of the measurement chip 90, the excitation light source 260 is generated. The light L to be blocked can be blocked before the fluorescent filter unit 262.

With reference to FIG. 1, the measurement part 14 is further demonstrated.
As shown in FIG. 1, the measurement unit 14 includes at least an excitation light source 260 as a light source, an optical filter (excitation filter) 720 for selecting only a desired excitation wavelength band from light emitted from the light source, An optical filter (fluorescence filter) 721 for selecting only a desired wavelength band of optical information from the measurement chip 90 and a dichroic mirror 751 for switching the optical path according to the difference in wavelength band between excitation light and optical information. At least one fluorescent filter unit 262, an objective lens 110 for guiding light emitted from the excitation light source 260 to the measurement chip 90 and collecting optical information obtained from the measurement chip 90, and an objective lens 110. A focus unit 265 having an autofocus function that can be moved in the optical axis direction, and an optical information from a measurement target A light receiving section 266 as a light detection unit for detecting a composed. As shown in FIG. 1, the fluorescent filter unit 262 and the light receiving unit 266 are fixed to the fluorescent incident unit 752.

  Furthermore, the measurement unit 14 includes a half mirror (not shown), and by switching between the half mirror and the fluorescent filter unit 262, the observation target is irradiated with a part of the light from the light source 260 and at the same time. By guiding a part of the reflected light from the light receiving unit 266 that is a light detecting unit, the shape and position information of the upper surface 90S of the measuring chip 90 and the well 200 formed on the upper surface 90S can be measured. The half mirror is an optical component that generally has a function of splitting incident light by 50% intensity, but in this embodiment, the light intensity of the spectrum does not necessarily need to be 50%. However, since the measurement accuracy improves as the light intensity incident on the light receiving unit 266 increases, it is preferable to use a branching mirror having a high light intensity split into the light receiving unit 266.

  In the fluorescence filter unit 262, only the wavelength band component of the excitation light is extracted from the light source 260 by the optical filter 720 (excitation filter), reflected (transmitted) by the dichroic mirror 751, and incident on the objective lens 110 for measurement. The chip 90 is irradiated. The light that returns from the measurement chip 90 as reflected light is reflected (transmitted) by the dichroic mirror 751 again, and therefore returns to the light source 260 side and is not propagated to the light receiving unit 266 side.

On the other hand, since fluorescence has a longer wavelength than the excitation light, only a wavelength band that is transmitted (reflected) through the dichroic mirror 751 and is detected by the optical filter 721 (fluorescence filter) is a detection unit (CCD camera or the like). Propagated to the light receiving unit 266.
When measuring the shape and position of the measurement chip surface, the well, and the tip of the collection unit by reflected light, only the half mirror is used instead of the fluorescent filter unit 262. When a half mirror is used in this way, the half mirror is placed at the location of the dichroic mirror 751 and the other two optical filters 720 and 721 are basically not required, but at a position corresponding to the excitation filter in the fluorescence filter unit. As the filter, it is more preferable to use a filter that selectively transmits a wavelength at which the fluorescence to be measured is less likely to fade.
As described above, measurement of position information by reflected light using a half mirror has an advantage that a light source common to fluorescence measurement can be used. This is not limited to a screening apparatus having a measurement function and a recovery function, and is a common advantage even in an apparatus that only measures.

  The plurality of objective lenses 110 and 264 of the measurement unit 14 shown in FIG. 1 can be rotated at a revolver type, for example, so that an objective lens having a necessary magnification can be positioned at a position below the measurement chip 90. The focus unit 265 moves and positions, for example, the objective lens 110 disposed in the lower position of the measuring chip 90 along the Z direction by operating the motor 265M according to a command from the control unit 100, for example. The focus adjustment of the objective lens 110 with respect to the fine particles M of the measurement chip 90 can be performed.

  As shown in FIG. 1, the measurement unit 14 includes a plurality of light sources 250 above the measurement chip 90 in addition to the light sources 260. The light source 250 irradiates light from two or more directions symmetrical with respect to an axis perpendicular to the upper surface 90S of the measurement chip 90. The shape and position information of the upper surface 90S of the measurement chip 90 and the well 200 formed on the upper surface 90S can be measured by the transmitted light from the measurement chip 90. In this measurement, a fluorescent filter is not required, but by selecting the light source wavelength and the amount of light so that a sufficient amount of light remains even if it passes through the dichroic mirror and the fluorescent filter, this can be done with the fluorescent filter unit placed in the optical path. There is an advantage that observation is possible without removing or switching.

  The measurement unit 14 has an autofocus function using the contrast value of the measurement data as an index, maintains the focus on the upper surface 90S of the measurement chip 90 by the autofocus by transmitted light or reflected light during measurement, and reflects the reflected light. By performing auto-focusing of the tip / end portion 142 of the suction / discharge capillary 140, the position of the tip portion 142 of the suction / discharge capillary 140 in the measurement visual field and the distance from the measurement chip surface can be measured.

  In the fine particle screening apparatus 1, the light information is fluorescence, and the position information of the well 200 of the measurement chip 90 by shape information measurement by reflected light or transmitted light and the fluorescence from the fine particles by optical information measurement by excitation light. Measurement and analysis are performed using both information, and after measuring fluorescence by irradiating excitation light, transmitted light measurement in the same visual field is performed. That is, the fine particle screening apparatus 1 analyzes only the portion of the position of the well 200 specified by the shape information in the fluorescence measurement data. At that time, if the shape information is measured first, the fluorescence of the fine particles to be measured fades, and accurate information cannot be obtained. Therefore, the fluorescence luminance is measured by reflected light or transmitted light. It is effective to measure earlier.

  Further, by irradiating the region including the plurality of wells 200 on the measurement chip 90 with the excitation light, fine particles existing in the region including the plurality of wells 200 are measured in one field of view. At this time, when correcting the fluorescence measurement value from the fine particles using the luminance distribution detected by the light receiving unit 266 that is the light detection unit when the excitation light is irradiated to the substantially autofluorescent homogeneous member, Pixel noise is removed by averaging processing of a plurality of pixels in the luminance distribution or by a frequency filter.

  As a method for acquiring correction data, for example, using a measuring chip 90 as a homogenous member of autofluorescence, a method of acquiring an excitation light intensity distribution in a state where only a liquid used for suspending fine particles is injected, Extracting the luminance distribution of the part other than the well 200 from the measurement result, and complementing this data to obtain the luminance distribution of the entire measurement visual field, and further the luminance of the part of the chip surface where the well is not formed There is a method to acquire information. Furthermore, from the measurement information including the fine particles, the result of reducing the influence of the fine particles on the luminance distribution of the entire measurement visual field by data processing can be used.

  It is also possible to obtain a luminance distribution of a portion where no well is formed by using a measuring chip as a homogenous member of autofluorescence.

  The target of correction is not only the intensity distribution (non-uniformity) of the excitation light irradiated in the measurement field of view, but also the non-uniformity of detection efficiency due to the transmission efficiency of each optical system such as a lens or filter. . These non-uniformities are highly dependent on the radial distribution of the excitation light itself and the radial distribution of optical components such as lenses that guide the excitation light and fluorescence. It is preferable to significantly remove high frequency components. Even if the original image is obtained from measurement information containing fine particles, the effect of fine particle brightness can be substantially eliminated during the process, so depending on the required accuracy, information for correction can be obtained in advance. Rather than doing so, it is possible to correct simultaneously with the measurement. As a processing method, it is conceivable to use data processing using averaging, frequency analysis, cluster analysis, or the like. These multiple methods should be selected according to the required correction accuracy.

  In FIG. 1, when the shutter of the shutter unit 261 is opened, the light of the excitation light source 260 is transmitted only through the excitation wavelength band by the fluorescence filter 720 of the fluorescence filter unit 262, reflected by the dichroic mirror 751, and transmitted through the objective lens 110. The measurement chip 90 is irradiated. As a result, when the fine particles M in the measurement chip 90 generate fluorescence, for example, the fluorescence passes through the dichroic mirror 751, passes through only the arbitrary wavelength band by the fluorescent filter 721, and is received by the light receiving unit 266. The signal-electrical signal is converted, and the fluorescence is image-analyzed by the image analysis unit 15.

  Next, using the fine particle screening apparatus 1 shown in FIG. 1, the total number of fine particles including the fine particles emitting fluorescence in the well of the measuring chip 90 are selected from the fine particles in the other wells. An example of a screening method for sucking and collecting on the collecting plate 80 will be described with reference to FIGS.

  FIG. 7A is a cross-sectional view showing a part of the measurement chip 90, and fine particles are not yet contained in the well 200 of the measurement chip 90. FIG. 7B shows a state in which a plurality of fine particles M are arranged in each well 200 of the measurement chip 90.

  FIG. 8A shows a case where a plurality of fine particles M in the well 200 are irradiated with light (irradiation light, excitation light) L, and the plurality of fine particles M emit fluorescence. An example in which M1 emits fluorescence is shown. FIG. 8B shows a state in which the suction / discharge capillary 140 approaches the respective wells 200 in order and sucks and collects all the fine particles M including the fine particles M1 emitting fluorescence with the maximum luminance. Show.

  FIG. 9 shows an example in which all the fine particles including the fine particles M1 emitting fluorescence with the maximum luminance are sucked from the well 200 of the measurement plate 90 and collected in the well 81 of the collection plate 80.

  As shown in FIG. 1, on the mounting surface 70 of the mounting table 40, a recovery table 80 and a fixture 120 for the measuring chip 90 are respectively arranged at predetermined positions. The fixture 120 is disposed above the objective lens 11. The suction / discharge capillary 140 is positioned on the measurement chip 90.

  The light from the excitation light source 260 is propagated to the objective lens 11 as described above, and is applied to the measurement chip 90 as light L. As a result, the plurality of fine particles M in the measurement chip 90 shown in FIG. 8A generate fluorescence, and the fluorescence of the plurality of fine particles M passes through the dichroic mirror 751 as described above. Only the wavelength band desired to be detected by the optical filter (fluorescence filter) 721 is received by the light receiving unit 266, converted into an optical signal-electric signal, and the fluorescence is image-analyzed by the image analysis unit 15.

  The image analysis unit 15 analyzes the fluorescence intensity of at least the maximum brightness fine particles M1 among the plurality of fine particles M in each well 200. By analyzing the measured shape information and optical information, data for confirming that at least one particle satisfying an arbitrary luminance condition set by the measurer is present in at least each well 200 is acquired. Based on the analysis result, the measurer collects fine particles satisfying the luminance condition from the well 200.

  Based on the result of the image analysis unit 15, the control unit 100 operates the first motor 55 in response to the command of the control unit 100 shown in FIG. Thus, the second table 52 is moved and positioned along the X direction, and the second motor 65 is operated to move and position the mounting table 40 along the Y direction. Then, when the control unit 100 shown in FIG. 6 instructs and operates the motor 251, the suction / discharge capillary 140 is lowered in the Z2 direction.

  As a result, the suction / discharge capillary 140 can be positioned at any one well by approaching each well 200 of the measurement plate 90 as shown in FIG. 8B.

  Thereafter, as shown in FIG. 9, when the control unit 100 commands and operates the suction pump 133, the suction / discharge capillary 140 causes the total number of fine particles including the fine particles M <b> 1 emitting fluorescence with the maximum luminance. A total number of fine particles M including the fine particles M1 are selectively sucked from the wells 200 having M separately from the fine particles of the other wells 200.

  Furthermore, when the control unit 100 shown in FIG. 6 instructs and operates the motor 251, the suction / discharge capillary 140 is raised in the Z1 direction. When the control unit 100 shown in FIG. 3 instructs and operates the first motor 55, the second table 52 is moved and positioned along the X direction, and the second motor 65 is operated, thereby mounting table. By moving and positioning 40 along the Y direction, the suction / discharge capillary 140 is relatively moved and positioned on an arbitrary well 81 of the recovery plate 80. Then, when the controller 100 shown in FIG. 6 instructs and operates the motor 251, when the suction / discharge capillary 140 is lowered in the Z <b> 2 direction, the distal end portion 142 of the suction / discharge capillary 140 is predetermined on the recovery plate 80. It approaches the well 81 at the position.

  The well 81 is preliminarily filled with a liquid LD such as a culture solution, and the tip 142 of the suction / discharge capillary 140 enters the liquid LD.

  By operating the suction pump 133 in reverse, the suction / discharge capillary 140 can discharge all the fine particles M including the fine particles M1 into the liquid in the well 81 at a predetermined position of the collection plate 80. As a result, the total number of fine particles M including the fine particles M 1 emitting fluorescence with the maximum luminance can be collected in the well 81 of the collection plate 80.

  The above-described suction and collection operations for all the fine particles including the fine particles M1 emitting fluorescence with the maximum luminance are performed in order for each well 200 of the measurement chip 90 in which the fine particles satisfying the collection conditions are stored. Do.

  In this way, the total number of fine particles including the fine particles M 1 emitting fluorescence in each well 200 is moved by the moving unit 16 in the X and Y directions and sucked by the collecting unit 13. By the operation of moving the discharge capillary 140 up and down in the Z direction, it is possible to recover from the well 81 at a predetermined position of the recovery plate 80 reliably and efficiently. That is, it is possible to efficiently search for and selectively recover target fine particles from a large number of fine particles.

  In the fine particle screening apparatus 1 of the present invention, the suction / discharge capillary 140 moves in the vertical direction (Z direction) with respect to the measurement chip 90 and approaches each well 200 of the measurement chip 90. . As a result, the tip 142 of the suction / discharge capillary 140 approaches each well 200 with certainty, and the total number of fine particles including the target fine particles M1 from the well 200 to the fine particles in the other wells. It can be collected by aspiration separately.

  FIG. 10 shows an example of a reference surface for height adjustment of the tip 142 of the suction / discharge capillary 140. This height matching reference surface 890 is set on the fixture 121, and the height from the surface of the measurement chip 90 is known. The reference surface 890 for height adjustment coincides with the inner surface 124 of the fixture 121. The objective lens 110 shown in FIG. 1 focuses on the reference surface 890 for height adjustment, and with the objective lens 110 held at that position, the collection unit 13 reaches a position where the tip 142 of the suction / discharge capillary 140 is in focus. Move. Thus, the distance from the upper surface 90S of the measurement chip 90 is determined, and the tip 142 of the suction / discharge capillary 140 is moved from the upper surface 90S of the measurement chip 90 by relative movement from the position coordinates of the recovery unit 13 at this time. The desired distance V is lowered to the position 891 along the Z2 direction to approach the upper surface 90S. As a result, the tip 142 of the suction / discharge capillary 140 can be reliably aligned along the Z direction using the height matching reference surface 890.

  FIG. 11 shows the recovery success rate when the fine particles are recovered by the syringe method using the suction / discharge capillary 140. The definition of the recovery success rate refers to the probability that all the fine particles in the well to be recovered are recovered by the suction / discharge capillary 140, and no single fine particle is recovered from the adjacent well.

  The number of trials is 10 under each condition, and suction is by stroke control. The amount of suction was set such that the recovery rate was maximized under each condition. The suction speed was constant. As the suction target, immobilized yeast cells in PBS (Phosphate buffered saline) were used. The well diameter is about 10 μm and the well pitch is about 30 μm. The approach distance of the suction / discharge capillary 140 to the measurement chip is four conditions of 10, 20, 30, 40 μm, and the inner diameter of the tip 142 of the suction / discharge capillary 140 is four conditions of 10, 20, 30, 40 μm. It was.

  Referring to FIG. 11, the recovery success rate is higher as the approach distance (μm) to the measurement chip is smaller, and the recovery success rate is higher as the inner diameter (μm) of the tip 142 of the suction / discharge capillary 140 is smaller.

  That is, the inner diameter of the tip 142 of the suction / discharge capillary 140 is preferably not more than twice the diameter of the well, and the approach distance to the measuring chip is preferably not more than the well pitch. By doing so, the suction / discharge capillary 140 can more reliably collect fine particles from the target well by the syringe method.

  The suction / discharge capillary 140 is recorded by recording the tip 142 of the suction / discharge capillary 140 in the measurement field of view and aligning the well side to be collected of the measurement chip 90 with the tip of the capillary by the moving unit 16. Align the center of the tip 142 with the center of the well. Thereby, the total number of fine particles in the well can be recovered more reliably.

  When the amount of suction is controlled when the fine particles are sucked by the suction / discharge capillary 140, the suction speed is constant or the speed is decreased stepwise or continuously during the suction with reference to the suction speed at the beginning of the suction. . That is, depending on the type of fine particles, the fine particles may be adsorbed on the wall surface of the well, and a suction force that exceeds this adsorption force is required. No suction is required. On the contrary, after that, it is necessary to reduce the suction force in order to prevent erroneous suction from the adjacent well. Thus, when sucking, the fine particles can be reliably sucked from the well by controlling the suction force.

  As shown in FIG. 9, the tip portion 142 of the suction / discharge capillary 140 can be prefilled with a suspension 500 that is an example of a liquid. Thus, if the liquid is not put in the tip part 142 in advance, the liquid flows from the tip part 142 of the suction / discharge capillary 140 by capillary action unless the airtightness of the recovery part 13 is perfect. The speed of the inflow of the liquid depends on the degree of airtightness of the recovery unit 13, but the final inflow amount depends only on the physical properties of the suction / discharge capillary 140 and the liquid and the diameter of the suction / discharge capillary 140 and flows in Enters the position of the weight and balance of the liquid. For this reason, unnecessary fine particles also flow in with the liquid. The liquid to be put in is preferably used for actually suspending the measurement fine particles, but is not limited thereto.

  The filling amount is preferably a balance position by capillary action, which is determined by the physical properties of the solution (viscosity / density), the affinity between the suction / discharge capillary 140 and the solution, and the inner diameter of the capillary, but there is an error in the filling amount. Even in this case, the inflow speed can be reduced, and the accuracy of the suction / discharge operation can be increased.

FIG. 12 shows a preferred shape example of the measuring chip 90.
The measurement chip 90 shown in FIG. 12 is a rectangular flat plate as described above and has the well region 200R, and is, for example, a resin molded product. For example, two positioning protrusions 720 protrude from one long side portion 710 of the measuring chip 90, and one positioning protrusion 721 protrudes from one short side portion 711, for example. Is formed. The protrusions 720 and 721 have, for example, a rectangular shape, the protrusion 720 has an abutting surface 720B, and the protrusion 721 has an abutting surface 721B. That is, the protrusions 720 and 721 are formed so as to protrude from the two side surfaces of the measuring chip 90, respectively.

The abutting surface 720B of the protrusion 720 abuts against the positioning portion 120F inside the fixture 120, and the abutting surface 721B of the projection 721 abuts against the positioning portion 120G inside the fixture 120.
As a result, the measuring chip 90 can be reliably positioned by abutting along the abutting direction GH against the positioning portions 120F and 120G inside the fixture 120, so that the positioning can be performed with good reproducibility. Improves. On the other hand, when no projection is used, the entire corresponding side surface of the measuring chip 90 is brought into contact for positioning by abutment with the fixture, but accuracy is ensured over the entire side surface. It is very difficult.

  Further, the display unit 750 of the measurement chip 90 is formed on the upper surface 90S side of the measurement chip 90. The display unit 50 has an arrow shape, for example, and faces the abutting direction GH. The display unit 50 may be provided on the upper surface 90S, the lower surface, or the outer peripheral portion of the measurement chip 90, and is a mark such as a pattern or notch that clearly indicates the orientation (front and back and direction) when the measurement chip 90 is set. .

Next, FIG. 13 shows that the warping of the measurement chip 90 is eliminated when the measurement chip 90 is fixed to the fixture 120.
FIG. 13A is an embodiment of the present invention, and FIG. 13B is a comparative example.
In FIG. 13A, the objective lens 110, the light receiving portion 266, and the fixture 120 are shown. The upper and lower surfaces of the measuring chip 90 are examples in which a low-order aspect is formed as a shape as typified by, for example, warping due to a temperature change of a material after release in molding.

  The upper surface 90S of the measuring chip 90 is applied to the inner surface 124 of the fixture 120, and the warping of the measuring chip 90 is corrected by applying a force FC from the lower surface 90D side of the measuring chip 90. That is, the region including the measurement area on the upper surface 90S side where the well is formed is pressed against the inner surface 124 which is a flat rigid plane of the fixture 120 by applying a force from the lower surface 90D which is the opposite surface. Correct 90 warpage. The parallelism of an arbitrary 2 mm square region of the measurement area with respect to the inner surface 124 that is a flat rigid plane is 10 μm or less (excluding minute undulations and protrusions such as the inclination of the well itself). Thereby, the positional accuracy of the well with respect to the optical system is improved, and focusing can be ensured in the entire region within the observation field.

  On the other hand, in the comparative example shown in FIG. 13B, there are small irregularities on the upper surface 90SN and the lower surface 90DN of the measuring chip 90N, and even if the upper surface 90SN of the measuring chip 90N is the fixing tool 120. Even if a force FC is applied from the lower surface 90DN side of the measuring chip 90N against the inner surface 124, the warping of the measuring chip 90 cannot be corrected, and within one observation field (for example, 2 mm square). Focus cannot be ensured throughout.

  Next, FIG. 14 shows the shift of the observation focus caused by the thickness variation of the measuring chip 90. FIG.

  14A to 14C, the distance between the upper surface of the measurement chip (the surface on which the well is formed) and the optical system are all the same, but the focal position varies depending on the thickness of the chip. Show. That is, in FIG. 14B, the focal point MP of the optical system 976 is accurately positioned on the upper surface 90S, whereas in FIG. 14A, the focal point MP is located above the upper surface 90S, and FIG. ) Are located on the lower side, and both are out of focus from the measurement target position. If the amount of deviation exceeds the depth of field, normal measurement is impossible. In the case of a general measurement optical system for securing an observation field of view of about 2 mm square, it is necessary to suppress the focus shift within a range of 10 μm, and an allowable thickness fluctuation amount is calculated as follows.

In FIG. 14, t is the thickness [mm] of the measuring chip 90, Δt is the thickness variation [mm] of the measuring chip 90, and L ₀ and L are focal lengths [mm] of the lens 110 </ b> B of the objective lens 110. , And ΔL is a focus shift [μm].

Here, the focal length L has the relationship of Formula 1.
L = L ₀ + t × (1-1 / n) (1)
From Equation 1,
ΔL = Δt × (1-1 / n)... Equation 2
Is obtained.
In order to make the focus shift within ± 5 μm (the focus shift width within one field of view is within 10 μm),
Δt ≦ 10 / (1-1 / n)... Formula 3
It becomes.

  When the measuring chip 90 is made of polystyrene (refractive index n = 1.59), Δt is within ± 13.5 μm and the thickness variation Δt is in order to make the focal shift ΔL within ± 5 μm. It is within 27 μm.

  As a result, even if the thickness of the measuring chip 90 varies within this range, it is possible to suppress the focal shift within an area of any 2 mm square within ± 5 μm. Can be prevented.

  FIG. 15 shows an error in the observation position due to a parallel shift between the upper surface 90S and the lower surface 90D of the measuring chip 90. FIG. In this fine particle screening device, the position of the well and the suction / discharge capillary is grasped and corrected based on the observation image, and the position recognition accuracy by the image requires ± 5 μm. Therefore, the distortion of the observation image is below this level. It is necessary to keep it down.

FIG. 15A illustrates an example in which the upper surface 90S and the lower surface 90D are parallel, and FIG. 15B illustrates an example in which the upper surface 90S and the lower surface 90D are not parallel. The observation target MJ is a well or the tip of a suction / discharge capillary. On the upper surface 90S on which at least the well of the measurement chip 90 is formed, the upper surface 90S and the lower surface 90D are used so that the position recognition accuracy within an arbitrary 2 mm square region included in the measurement area of the optical system is within ± 5 μm. The inclination angle (displacement from parallel) φ is less than (5 × 10 ⁻⁶ ) / {t (1-1 / n)} [rad] (t is the average thickness of the measuring chip [ mm] and n indicate that the refractive index of the material of the measurement chip is required.

  Δ indicates the deviation of the observation position of the observation object MJ, t indicates the average thickness of the measurement chip 90, n indicates the refractive index of the material of the measurement chip 90, and φ indicates the inclination angle of the lower surface 90D.

The deviation Δ of the observation position of the observation object MJ can be expressed by Equation 4.
Δ = t · tan [φ−sin ⁻¹ {(sin φ) / n}] ≈t · φ · (1-1 / n) Equation 4
In order to make Δ ≦ 5 μm,
φ ≦ (5 × 10 ⁻⁶ ) / {t (1-1 / n)} [rad] Expression 5
It becomes.

  Thereby, the position recognition accuracy can be within ± 5 μm at least in the entire observation field (for example, 2 mm square), and the observation target in the entire observation field (for example, 2 mm square) can be reduced. The position recognition accuracy is improved.

  FIG. 16 shows an example of the relationship between the inclination angle φ for satisfying the position recognition accuracy within ± 5 μm and the deviation Δ of the observation position, the chip thickness t is 1 mm, and the refractive index n corresponds to polystyrene. This is the case of 1.59. Since the deviation Δ (μm) of the observation position is within 5 μm, the variation in the tilt angle is 0.8 degrees or less.

  In the embodiment of the present invention, the measurement chip 90 has an area in which a well is formed having a width of 10 mm square or more, and an appropriate part or all of the surrounding area not used for measurement is the fixture 120. Is provided. An appropriate size from the lower surface side where the well of the measuring chip 90 is not formed in a direction to be pressed against a reference surface (corresponding to the inner surface 124) that can be regarded as a substantially rigid body included in one plane of the fixture 120. By applying the force FC, the flatness of an arbitrary 2 mm square region included in the entire region where the well is formed with respect to the reference surface (corresponding to the inner surface 124) can be 10 μm or less. For this reason, positioning with high accuracy can improve positioning workability and prevent defocusing and deviation of the recognition position.

By the way, this invention is not limited to the said embodiment, A various modified example is employable.
For example, in the embodiment of the present invention, biological cells are exemplified as target fine particles, but the present invention is not limited to this, and other types of fine particles may be used.

  Further, as shown in FIG. 3, one collection plate 80 is arranged on the mounting plate 40, but a plurality of collection plates may be arranged side by side.

  As the drive system for linear movement of the moving unit 16, a rotary electric motor, a feed screw, and a guide rail are used, but a linear motor or the like can be used instead of the rotary electric motor.

  The cross-sectional shape of the well 81 of the recovery plate 80 and the cross-sectional shape of the well 200 of the measuring chip 90 are not limited to the illustrated example, and other shapes such as a hemispherical shape can also be adopted.

  The fine particle screening apparatus of the present invention is a field that requires testing, analysis and analysis of biopolymers of genes, immune systems, proteins, amino acids, and sugars, such as engineering, food, agriculture, fishery processing, etc. It can be applied to various fields such as pharmaceutical field, hygiene, health, immunity, plague, genetic field such as heredity, and scientific field such as chemistry or biology.

It is a front view which shows preferable embodiment of the screening apparatus of the fine particle of this invention. It is a perspective view which shows the screening apparatus of the fine particle of FIG. It is a perspective view which shows the mounting table, collection | recovery plate, and measurement chip | tip mounted on the moving part and the moving part. It is a figure which shows the mounting table, the collection | recovery plate, and the chip | tip for a measurement. It is a figure which shows the example of a shape of the measuring chip and the holding member of this measuring chip. It is a perspective view which shows the structural example of a collection | recovery part. FIG. 7A is a cross-sectional view showing a part of the measurement chip, in which fine particles are not yet contained in the well of the measurement chip. FIG. 7B is a diagram showing a state where fine particles M are inserted into the wells of the measurement chip. FIG. 8A is a diagram showing an example in which the light L is irradiated to the fine particles M in the well and a part of the fine particles M emits fluorescence. FIG. 8B is a diagram illustrating a state where the suction / discharge capillary approaches the selected well and sucks and collects all the fine particles. FIG. 9 is a diagram showing an example in which all the fine particles including the fluorescent fine particles M1 are sucked from the wells of the measurement plate and collected in the wells of the collection plate. It is a figure which shows the example of the reference surface for capillary height adjustment. The collection success rate when collecting fine particles with a capillary is shown. It is a figure which shows the preferable example of a shape of the chip | tip for a measurement. It is a figure which shows that the curvature of a measurement chip is eliminated when the measurement chip is fixed to a fixture. It is a figure shown about the focus shift by the fluctuation | variation of the thickness of the chip | tip for a measurement. It is a figure about the error of the observation position by the parallel shift of the upper surface and lower surface of a measurement chip. It is a figure which shows the example of relationship between inclination-angle (phi) and the observation position shift (DELTA) for satisfy | filling the position recognition accuracy within +/- 5micrometer.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Fine particle screening apparatus 11 Base 12 Support part 13 Collection | recovery part 14 Measurement part 15 Image analysis part 16 Moving part 19 Cover 30 Support stand 40 Mounting table 51 1st table 52 2nd table 55 1st motor 56 Feed screw 57 Nut 65 Second motor 66 Feed screw 67 Nut 70 Mounting surface 80 Recovery plate 81 Well of recovery plate 90 Measurement chip 90S Upper surface 90D Lower surface 100 Control unit 110 Objective lens 120 Fixing tool for measurement chip 130 Operation unit 133 Suction pump 140 Suction / discharge Capillary 142 Suction / discharge capillary tip 200 Measuring chip well 260 Excitation light source 251 Motor 252 Feed screw 253 Nut 261 Shutter unit 262 Fluorescence filter unit 264 Objective lens 266 Receiving Light part 500 Suspension 720, 721 Protrusion 750 Display part LD Liquid L Light (excitation light, irradiation light)
X 1st direction Y 2nd direction Z 3rd direction (vertical direction)
CL Reference surface on top of measurement chip

Claims (6)

A fine particle screening device for performing a search for fine particles using light information emitted from the fine particles as a label, and selectively acquiring the searched fine particles,
A measuring chip provided with a plurality of wells formed of a material that transmits light and having a size capable of accommodating at least one of the fine particles;
A fixture for fixing the measurement chip;
A measurement unit that obtains an optical signal of the fine particles by irradiating light to the fine particles accommodated in the measurement chip and the measurement chip; and
The measuring chip is
A protrusion provided on a side surface of the measurement chip in order to be positioned in a direction parallel to the upper and lower surfaces of the measurement chip by making contact with the abutting portion of the fixture;
A fine particle screening apparatus, comprising: a display unit that is formed on the measurement chip and clearly indicates a direction when the measurement chip is set on the fixture.   2. The fine particle screening apparatus according to claim 1, wherein the protrusion is formed so as to protrude from a side surface of the measurement chip.   In the measuring chip, the area where the well is formed is 10 mm square or more, and a part or all of the area around the well is fixed by the fixture, and the well is not formed. 3. The flatness of an arbitrary 2 mm square in the area where the well is formed is set to 10 μm or less by applying a force having an appropriate magnitude from the lower surface side. Fine particle screening device.   The thickness variation Δt in an arbitrary 2 mm square region included in the measurement area in which the well of the measurement chip is formed is 10 / (1-1 / n) [μm] or less (n is the measurement chip) The screening apparatus for fine particles according to claim 3, wherein the refractive index is a refractive index of the material ratio. The fluctuation of the inclination angle φ formed by the upper surface and the lower surface in an arbitrary 2 mm square region included in the measurement area where the well of the measurement chip is formed is (5 × 10 ⁻⁶ ) / {t ( 5/1 / n)} [rad] or less (t is an average thickness [mm] of the measurement chip, and n is a refractive index of a material of the measurement chip). Fine particle screening device. A fine particle screening method for searching for fine particles using light information emitted from the fine particles as a label, and selectively acquiring the searched fine particles, wherein the measuring chip is formed of a material that transmits light A plurality of wells having a size capable of storing at least one of the fine particles, the measurement chip is fixed to a fixture, and a measurement unit includes the measurement chip and the measurement When acquiring the optical signal of the fine particles by irradiating the fine particles contained in the chip with light,
When positioning the projection of the measuring chip by bringing it into contact with the abutting portion of the fixture, the projection is set with respect to the fixture based on a display section that clearly indicates the orientation to be set with respect to the fixture. A screening method for fine particles, characterized by:

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