JP2009300454A - Imaging flow site meter - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an imaging flow site meter capable of obtaining a transmitted light image with good contrast and of computing the size of a cell from the transmitted light image at high speed by simple image processing. <P>SOLUTION: The imaging flow site meter is constituted by comprising a sheath flow cell which surrounds cell containing liquid containing the cell by sheath liquid and forms a sample flow, a light source which illuminates the sample flow, a first imaging part which picks up the transmitted image of the rear pin of the cell in the sample flow, a second imaging part which picks up the fluorescence image of the cell in the sample flow, a display part, and a control part which computes the size of the cell based on the transmitted light image of the rear pin of the cell and displays the computed size of the cell and the fluorescence image of the cell on the display part at least. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to an imaging flow cytometer, and more particularly to a flow cytometer having a function of imaging particles flowing in a flow cell.

As a background art of the present invention, a liquid containing test particles is flowed through a flow cell to form a sample flow, the sample flow is illuminated with laser light, and two fluorescent images and two transmitted light images of the illuminated particles are provided. An image pickup unit (image sensor) is known (see, for example, Patent Document 1).

JP 2003-083960 A

However, in such a conventional apparatus, when the test particle is a transparent object such as a cell having a higher refractive index than the surrounding medium, the transmitted light image is almost lost, and a transmitted light image with good contrast is obtained. It was difficult to get. The present invention has been made in consideration of such circumstances, and a transmitted light image with good contrast can be obtained by setting the focus of the imaging system of the transmitted light image appropriately deviated from the accurate focus. is possible, in a simple image processing, the size of the cells from the transmission light images that provide calculable imaging flow cytometer at a high speed.

The present invention includes a first imaging and the sheath flow cell for forming a sample flow wrapped cell-containing liquid containing cells in the sheath fluid, a light source for illuminating the sample stream, the pins of the transmitted light image after cell sample stream One imaging unit, a second imaging unit that captures a fluorescence image of a cell in the sample flow , a display unit, and a cell size is calculated based on a transmitted light image of the back pin of the cell, and at least the calculated cell size a control unit for displaying on the display unit the fluorescence image and cells that provide Bei obtain imaging flow cytometer.

In another aspect, the present invention provides a sheath flow cell that forms a sample flow by wrapping a cell- containing liquid containing cells in a sheath liquid, a light source that illuminates the sample flow, and a transmitted light image of the cells in the sample flow A first imaging unit that images the first imaging system, a first optical system that projects a cell image in the sample flow onto the imaging surface of the first imaging unit , and a sample that has a focal point far from the imaging surface of the first imaging unit. A second imaging unit that captures a fluorescent image of a cell in flow, and a second optical system that has a focal point on the imaging surface of the second imaging unit and projects an image of a cell in the sample stream on the imaging surface of the second imaging unit When a display unit, based on the cells of the transmitted light image to calculate the size of the cell, and a control unit for displaying the fluorescence image size and cells of cells at least calculation on the display unit, an imaging flow cytometer equipped with To provide .

Is preferred flow path cross section of the flat type for sheet Sufuroseru but the cross-section of the flow channel can also be used a circular or rectangular. As the light source, a white lamp , a laser , a strobe flash lamp, a pulse laser, or the like can be used. A CCD camera , a video camera, or the like is suitable for the first and second imaging units. In particular, it is preferable that an optical amplifying element, for example, an image intensifier is attached to the second imaging unit that captures a fluorescent image. . In the present invention, the “rear pin” refers to a state in focus after the subject.

Light source, a first light source for emitting white light, made from the second light source for emitting a laser beam, the first imaging section images the cells illuminated by the first light source, the second imaging unit, a second You may make it image the cell illuminated with the light source. The first light source may be arranged such that the first imaging unit images particles by transmitted illumination, and the first light source is arranged so that the second imaging unit images particles by epi-illumination. The light source may be a pulsed light source.

A control section includes a displacement portion for displacing the sheath flow cell along the optical axis of the first imaging unit, and a calculation unit for calculating an average luminance of the cells of the transmitted light image, a predetermined value the average luminance is calculated by the arithmetic unit a drive control unit for driving the displacement portion so that it may be provided with.

Imaging flow cytometer according to the present invention, since the first imaging unit captures the transmitted light image of the pin after cell, cell fraction images appear wrapped in black Becke line. Therefore, it is possible to obtain a transmitted light image of a cell with good contrast by the first imaging unit. Furthermore, the imaging flow cytometer according to the present invention makes it easy to extract the outline of a cell image by using a transmitted light image of a cell with good contrast, and can calculate the cell size at high speed with simple image processing. is there.

BRIEF DESCRIPTION OF THE DRAWINGS It is structure explanatory drawing which shows 1st Example of this invention. It is a timing chart of the signal of 1st Example of this invention. It is a flowchart which shows operation | movement of 1st Example of this invention. It is a structure explanatory drawing which shows 2nd Example of this invention. FIG. 6 is an explanatory diagram showing a configuration of a third embodiment of the present invention.

EXAMPLES Hereinafter, the present invention will be described in detail based on examples shown in the drawings. This does not limit the present invention.
First Embodiment FIG. 1 is an explanatory diagram showing the construction of a first embodiment of the present invention. As shown in the figure, a sheath flow cell (hereinafter referred to as a flow cell) 1 is a cylindrical cell having an elongated rectangular cross section, and is formed of a transparent material such as glass or plastic. A sample liquid and a sheath liquid are introduced into the flow cell 1, and the sample liquid is wrapped in the sheath liquid and converted into a flat sample flow 5 of about 1 μm × 2000 μm.

  In this example, for K562 or HL-60 cells, 200 μL of a sample solution in which 2000 cells / μL of cells are suspended is prepared per sample, and the sample solution is allowed to flow through the flow cell 1 at a flow rate of 2 to 5 cm / sec. I have to. In addition, the cell nucleus is fluorescently stained in advance with Hochest 33342.

The transmission image imaging system includes a first illumination optical system for illuminating cells in the sample stream 5 and a first light receiving optical system (first optical system) for receiving a light image from the cells in the sample stream 5. Consists of. The first illumination optical system includes a first light source 2, a condenser lens 3 and a reflection mirror 4. The first light receiving optical system (first optical system) includes an objective lens 6, a dichroic mirror 7, a fluorescent filter 8, a half mirror 9, a reflection mirror 10, an imaging lens 11, and a first imaging unit 12.
On the other hand, the imaging system of the fluorescence image includes a second illumination optical system for illuminating the cells of the sample stream 5 and a second light receiving optical system (second optical system) for receiving a light image from the cells of the sample stream 5. Become. The second illumination optical system includes a second light source 13, a condenser lens 14, a dichroic mirror 7 and an objective lens 6. The second light receiving optical system (second optical system) includes an objective lens 6, a dichroic mirror 7, a fluorescent filter 8, a half mirror 9, an imaging lens 15, a gated image intensifier 16, and a second imaging unit 17. Is done.

  Here, the first light source 2 is a pulse light source set to perform Keller illumination via the condenser lens 3, and a strobe flash lamp that emits white light (wavelength: 500 nm to 650 nm) is used for this. The second light source 17 is a pulsed light source set to perform critical illumination via the condenser lens 14, and a UV pulse laser that emits ultraviolet light having a center wavelength of 365 nm is used for this. As the dichroic mirror 7, a mirror that transmits light having a wavelength of 400 nm or more and reflects light having a wavelength shorter than 400 nm is used. The fluorescent filter 8 transmits only light having a wavelength of 450 nm ± 50 nm. In addition, a half mirror 9 is used that transmits half the amount of incident light and reflects the rest. The first and second imaging units 12 and 17 are composed of CCD cameras (video cameras).

  Next, the control unit 21 includes a signal processing unit 24, a display unit 30, and a displacement unit 29. The displacement unit 29 includes a stepping motor 23 and a linear movement mechanism 22. The linear movement mechanism 22 decelerates the rotational output of the stepping motor 23 and converts it into linear motion, thereby displacing the flow cell 1 in the optical axis direction (arrow A or B direction) of the imaging system. That is, when a driving pulse for one step is applied to the stepping motor 23, the flow cell 1 is displaced by 0.05 μm.

  The signal processing unit 24 includes a calculation unit 25, a storage unit 26, a drive control unit 27, and an imaging control unit 28, and is integrally configured by a microcomputer. The calculation unit 25 processes image data obtained from the first and second imaging units 12 and 17 and performs calculations necessary for analysis. The storage unit 26 stores the image data processed by the calculation unit 25 and the calculated calculation result. The drive control unit 27 applies a drive pulse to the stepping motor 23 based on the calculation result of the calculation unit 25 to drive-control the displacement unit 29. The imaging control unit 28 controls the light emission operation of the first and second light sources 2 and 13, the gate opening / closing operation of the intensifier 16, the imaging operation of the first and second imaging units 12 and 17, and the like.

  The focal point of the first light receiving optical system is adjusted to a position farther than the imaging surface of the first imaging unit 12, so that a transmitted light image surrounded by a black Becke line can be obtained, and between the imaging lens 11 and the imaging unit 12. The optical path length is set. Further, the optical path length between the imaging lens 15 and the second imaging unit 17 is set so that the focal point of the second light receiving optical system matches the imaging surface of the second imaging unit 17, and a fluorescent image with good contrast is obtained. Here, the focal point of the objective lens 6 is set so as to match the central axis of the sample flow 5.

Next, the operation in such a configuration will be described.
2 shows signals output from the imaging control unit 28 of the control unit 1, that is, pulse signals S1 and S3 for turning on the second light source 13 and the first light source 2, respectively, and a pulse signal for opening the gate of the image intensifier 16. It is a timing chart of S3.
First, when the sample flow 5 is formed in the flow cell 1, the second light source 13 emits light having a wavelength of 365 nm by the pulse signal S1 output from the imaging control unit 28. The light emitted from the second light source 13 passes through the condenser lens 14 and is reflected by the dichroic mirror 7 toward the objective lens 6 to illuminate the cells in the sample flow 5 through the objective lens 6. Thereby, fluorescence having a wavelength of 400 to 500 nm is excited from the fluorescence-stained nucleus in the cell, and image in is performed through the imaging lens 6, the dichroic mirror 7, the fluorescent filter 8, the half mirror 9, and the imaging lens 15. Reach the tensifier 16. At this time, since the gate of the image intensifier 16 is opened as shown in FIG. 2 by the pulse signal S2 output from the imaging control unit 28, the fluorescence image of the nucleus is applied to the photocathode of the image intensifier 16. Imaged and amplified. The amplified fluorescence image is captured by the second imaging unit 17 and sent to the signal processing unit 24 of the control unit 21 as fluorescence image data.

  As shown in FIG. 2, when the pulse signal S2 for opening the gate of the image intensifier 16 is turned off and the gate is closed, the pulse signal S3 is then output from the imaging control unit 28, and the first light source 2 is turned on. Emits white light. The outgoing light of the first light source 2 illuminates the cells in the sample flow 5 via the condenser lens 3 and the reflection mirror 4. And the transmitted light which permeate | transmitted the cell injects into the 1st imaging part 12 via the dichroic mirror 7, the fluorescence filter 8, the half mirror 9, the reflective mirror 10, and the imaging lens 11, and is 1st as a transmitted light image of a cell. The image is captured by the imaging unit 12 and transmitted to the signal processing unit 24 of the control unit 21 as transmitted light image data. Such imaging operations by the first and second imaging units 12 and 17 are repeated 30 times per second. As a result, a transmitted light image and a fluorescent image of 30 frames each are obtained per second. That is, 30 sets of transmitted light images and fluorescent images are obtained per second. Note that the time difference T between the pulse signals S1 and S3 shown in FIG. 2 is set to 20 μsec or less so that the positional deviation between the transmitted light image and fluorescent image of the same cell does not increase in each set of images.

Next, the control processing operation of the control unit 21 will be described using the flowchart of FIG.
When each of the first and second imaging units 12 and 17 completes imaging of each frame (step S1), first, processing for tracking the outline of the cell in the transmitted light image is performed, and the cell image is cut out (step S2). ). The average luminance Ia of the cut cell image is calculated and stored in the storage unit 26 (step S3), and the area S and the perimeter L of the cell image are calculated (step S4). Further, from the area S and the peripheral length L, the cell size (equivalent circle diameter) and the cell roundness (circularity) are calculated.

  Next, the cell image in the fluorescence image is cut out and stored in the storage unit 26 together with the above S and L (step S5). Further, the size of the cell nucleus (equivalent circle diameter) and the roundness of the cell nucleus (circularity) are calculated from the area S and the peripheral length L. Next, P = K (Is−Ia) is calculated in order to compare the average luminance Ia of the transmitted light image of the cell with the predetermined value Is (step S6). Here, K is a constant.

  Then, it is determined whether or not P is within a predetermined range ε, that is, whether or not | P | ≦ ε (step S7). If | P | ≦ ε, when the time of 1/30 seconds elapses from step S1 (step S11), the next frame is imaged. When | P | ≦ ε is not satisfied in step S7, and P> ε (step S8), a forward rotation drive pulse for one step is output from the drive control unit 27 of FIG. 1 to the stepping motor 23, and the flow cell. 1 moves in the direction of arrow A so as to move away from the objective lens 6. If P> ε is not satisfied in step S8, that is, if P <−ε, the reverse rotation drive pulse for one step is output from the drive control unit 27 to the stepping motor 23, and the flow cell 1 is sent to the objective lens 6. Move in the direction of arrow B to approach.

As described above, since the focus of the first light receiving optical system is adjusted to a position farther from the imaging surface of the first imaging unit 12, a cell image having a high contrast which is a transmitted light image wrapped in a black Becke line is obtained. It is done. By using a cell image having a high contrast, the outline of the cell image can be easily extracted, and the size and roundness of the cell image can be calculated at high speed by simple image processing.
Further, by using the size of the cell nucleus obtained from the fluorescence image of the cell, the effect of the drug can be confirmed by capturing the change in the size of the cell nucleus caused by the drug such as an anticancer drug.
In this embodiment, as described above, the focal point of the fluorescent imaging system is set so as to be aligned with the central axis of the sample flow 5, and the focal point of the transmitted light imaging system is fluorescent so that the transmitted light image of the cell is surrounded by a black Becke line. It is set farther than the focal point of the image pickup system. However, if the cell row flowing in the sample flow 5 is deviated from the central axis of the sample flow 5 for some reason, the position of the imaging target with respect to the focus of the transmitted light image capturing system and the fluorescence image capturing system is shifted, so that the desired image is Cannot get good contrast.
Therefore, in the present invention, as described above, the displacement of the cell row is detected from the average luminance of the cells in the transmitted light image (the density of the cell image), and the flow cell 1 is moved correspondingly, so that the imaging target is always in the normal position. The correction is made in the direction of.
In this way, when imaging for one sample is completed, various data stored in the storage unit 26 are displayed on the display unit 30 as necessary.

Second Embodiment FIG. 4 is an explanatory diagram showing the construction of a second embodiment of the present invention. In the figure, the same reference numerals are assigned to the same elements as those in the first embodiment (FIG. 1).
In FIG. 4, the transmission image capturing system includes a first illumination optical system for illuminating cells in the sample stream 5 and a first light receiving optical system (first array) for receiving a light image from the cells in the sample stream 5. 1 optical system). The first illumination optical system includes a first light source 2, a condenser lens 3 and a reflection mirror 4. The first light receiving optical system (first optical system) includes an objective lens 6, a dichroic mirror 7 a, an imaging lens 11, and a first imaging unit 12.
On the other hand, the imaging system of the fluorescence image includes a second illumination optical system for illuminating the cells of the sample stream 5 and a second light receiving optical system (second optical system) for receiving a light image from the cells of the sample stream 5. Become. The second illumination optical system includes a second light source 13, a beam expander 18, dichroic mirrors 7 and 7 a, and an objective lens 6. The second light receiving optical system (second optical system) includes the objective lens 6, the dichroic mirrors 7 a and 7, the fluorescent filter 8, the half mirror 9, the imaging lens 15, the gated image intensifier 16, and the second imaging unit 17. Consists of Here, the dichroic mirror 7a reflects light having a wavelength of 500 nm or less and transmits light having a wavelength greater than 500 nm.

  The focal point of the first light receiving optical system is adjusted to a position farther than the imaging surface of the first imaging unit 12, so that a transmitted light image surrounded by a black Becke line can be obtained, and between the imaging lens 11 and the imaging unit 12. The optical path length is set. Further, the optical path length between the imaging lens 15 and the second imaging unit 17 is set so that the focal point of the second light receiving optical system matches the imaging surface of the second imaging unit 17, and a fluorescent image with good contrast is obtained. Here, the focal point of the objective lens 6 is set so as to match the central axis of the sample flow 5.

  Next, the operation in such a configuration will be described. FIG. 2 is a timing chart of signals output from the imaging control unit 21, that is, pulse signals S1 and S3 for turning on the second light source 13 and the first light source 2, respectively, and a pulse signal S3 for opening the gate of the image intensifier 16. It is.

  First, when the sample flow 5 is formed in the flow cell 1, the second light source 13 emits light having a wavelength of 365 nm by the pulse signal S1 output from the control unit 21. The light emitted from the second light source 13 passes through the beam expander 18 and is reflected by the dichroic mirror 7 toward the dichroic mirror 7a. Illuminate. Thereby, fluorescence having a wavelength of 400 to 500 nm is excited from the fluorescence-stained nucleus in the cell, and image-in is performed through the imaging lens 6, the dichroic mirror 7 a, the dichroic mirror 7, the fluorescence filter 8, and the imaging lens 15. Reach the tensifier 16. At this time, the gate of the image intensifier 16 is opened as shown in FIG. 2 by the pulse signal S2 output from the imaging control unit 21, so that the fluorescence image of the nucleus is applied to the photocathode of the image intensifier 16. Imaged and amplified. The amplified fluorescence image is captured by the second imaging unit 17 and sent to the control unit 21 as fluorescence image data.

  As shown in FIG. 2, when the pulse signal S2 for opening the gate of the image intensifier 16 is turned off and the gate is closed, the control unit 21 outputs a pulse signal S3, and the first light source 2 emits light. To do. The outgoing light of the first light source 2 illuminates the cells in the sample flow 5 via the condenser lens 3 and the reflection mirror 4. The transmitted light that has passed through the cell is incident on the first imaging unit 12 via the objective lens 6, the dichroic mirror 7 a, and the imaging lens 11. Thereby, the transmitted light image of the cell is captured by the first imaging unit 12 and transmitted to the control unit 21 as transmitted light image data. Such imaging operations by the first and second imaging units 12 and 17 are repeated 30 times per second. As a result, a transmitted light image and a fluorescent image of 30 frames each are obtained per second. That is, 30 sets of transmitted light images and fluorescent images are obtained per second. The obtained fluorescence image data and transmitted light image data are processed as shown in FIG. 3 by the control unit 21 as in the first embodiment, and the position of the flow cell 1 is similarly driven and controlled.

Third Embodiment FIG. 5 is an explanatory diagram showing the configuration of a third embodiment of the present invention. In the figure, the same reference numerals are assigned to the same elements as those in the first embodiment (FIG. 1). In the third embodiment, the first light source 2 in the first embodiment is omitted.
The transmission image imaging system includes a first illumination optical system for illuminating cells in the sample stream 5 and a first light receiving optical system (first optical system) for receiving a light image from the cells in the sample stream 5. Consists of. The first illumination optical system includes the second light source 13, the condenser lens 14, and the reflection mirror 4. The first light receiving optical system (first optical system) includes an objective lens 6, a dichroic mirror 7, a fluorescent filter 8, a reflection mirror 10, an imaging lens 11, and a first imaging unit 12.
On the other hand, the imaging system for the fluorescence image includes a first illumination optical system for illuminating the cells in the sample stream 5 and a second light receiving optical system (second optical system) for receiving a light image from the cells in the sample stream 5. Become. The first illumination optical system includes the second light source 13, the condenser lens 14, and the reflection mirror 4. The second light receiving optical system (second optical system) includes an objective lens 6, a dichroic mirror 7, a fluorescent filter 8, an imaging lens 15, an image intensifier 16 with a gate, and a second imaging unit 17.

  The focal point of the first light receiving optical system is adjusted to a position farther than the imaging surface of the first imaging unit 12, so that a transmitted light image surrounded by a black Becke line can be obtained, and between the imaging lens 11 and the imaging unit 12. The optical path length is set. Further, the optical path length between the imaging lens 15 and the second imaging unit 17 is set so that the focal point of the second light receiving optical system matches the imaging surface of the second imaging unit 17, and a fluorescent image with good contrast is obtained. Here, the focal point of the objective lens 6 is set so as to match the central axis of the sample flow 5.

Next, the operation in such a configuration will be described.
First, when the sample flow 5 is formed in the flow cell 1, the second light source 13 emits light having a wavelength of 365 nm by the pulse signal output from the control unit 21. The light emitted from the second light source 13 passes through the condenser lens 14 and is reflected by the reflection mirror 4 to transmit and illuminate the cells in the sample flow 5. Thereby, fluorescence having a wavelength of 400 to 500 nm is excited from the fluorescence-stained nucleus in the cell, and the excited fluorescence passes through the imaging lens 6, the dichroic mirror 7, the fluorescence filter 8, and the imaging lens 15. The image intensifier 16 is reached. At this time, the gate of the image intensifier 16 is opened by a pulse signal output from the control unit 21, and a nuclear fluorescence image is formed on the photocathode of the image intensifier 16 and amplified. The amplified fluorescence image is captured by the second imaging unit 17 and sent to the control unit 21 as fluorescence image data.

  On the other hand, the transmitted light that has passed through the cell is reflected by the dichroic mirror 7 and enters the first imaging unit 12 via the reflecting mirror 10 and the imaging lens 11, and a transmitted light image of the cell is captured by the first imaging unit 12. Then, it is sent to the control unit 21 as transmitted light image data. Such imaging operations by the first and second imaging units 12 and 17 are repeated 30 times per second. As a result, a transmitted light image and a fluorescent image of 30 frames each are obtained per second. That is, 30 sets of transmitted light images and fluorescent images are obtained per second. The obtained fluorescence image data and transmitted light image data are processed as shown in FIG. 3 by the control unit 21 as in the first embodiment, and the position of the flow cell 1 is similarly driven and controlled.

  As described above, in the first to third embodiments, the focal point of the first light receiving optical system (first optical system) is set to be far from the imaging surface of the first imaging unit 12, but the present invention is based on this. It is not limited. For example, the focal point of the first light receiving optical system (first optical system) is set on the imaging surface of the first imaging unit 12, and the focal point of the objective lens 6 is set so as to be far from the central axis of the sample flow 5. You may do it. In this case, the second light receiving optical system has a configuration including a second objective lens different from the objective lens 6 of the first light receiving optical system, and the focus of the second objective lens is the sample stream 5. The center axis is preferred.

This application is a patent application subject to Article 19 of the Industrial Technology Strengthening Act .

DESCRIPTION OF SYMBOLS 1 Flow cell 2 1st light source 3 Condenser lens 4 Reflection mirror 5 Sample flow 6 Objective lens 7 Dichroic mirror 8 Fluorescence filter 9 Half mirror 10 Reflection mirror 11 Imaging lens 12 1st imaging part 13 2nd light source 14 Condenser lens 15 Imaging lens DESCRIPTION OF SYMBOLS 16 Image intensifier with a gate 17 2nd image pick-up part 18 Beam expander 21 Control part 22 Linear movement mechanism 23 Stepping motor 24 Signal processing part 25 Calculation part 26 Storage part 27 Drive control part 28 Imaging control part 29 Displacement part 30 Display part

Claims (11)

A sheath flow cell for forming a sample flow wrapped cell-containing liquid containing cells in the sheath fluid,
A light source for illuminating the sample stream;
A first imaging unit that images a transmitted light image of a back pin of a cell in a sample flow;
A second imaging unit that captures a fluorescence image of cells in the sample stream ;
A display unit;
Based on the transmitted light image of the pin after the cells were calculated size of the cell, at least the calculated cell size and cell fluorescence image Bei obtain imaging flow cytometer and a control unit for displaying on the display unit of the. A sheath flow cell for forming a sample flow wrapped cell-containing liquid containing cells in the sheath fluid,
A light source for illuminating the sample stream;
A first imaging unit that images a transmitted light image of a cell in a sample flow;
Focus is in a position farther than the imaging surface of the first imaging unit, a first optical system you project an image of cells in the sample stream to an imaging plane of the first imaging unit,
A second imaging unit that captures a fluorescence image of cells in the sample stream;
Focus is on the imaging surface of the second imaging unit, a second optical system you project an image of cells in the sample stream to an imaging plane of the second imaging unit,
A display unit;
Based on the cell of the transmitted light image to calculate the size of the cell, at least the calculated cell size and cell fluorescence image Bei obtain imaging flow cytometer and a control unit for displaying on the display unit of the. The imaging flow cytometer according to claim 1, wherein the control unit includes a storage unit that stores a transmitted light image of the cell and a fluorescent image of the cell. The control unit cuts out a cell image from the transmitted light image, calculates an area and a perimeter of the cut out cell image, and calculates a cell size based on the calculated area and perimeter. The imaging flow cytometer according to one. The cell nucleus of the cell is stained with a fluorescent dye, and the control unit cuts out the cell image from the fluorescence image, calculates the area and perimeter of the cut out cell image, and based on the calculated area and perimeter, the cell nucleus of the cell The imaging flow cytometer according to claim 1, which calculates a size. Light source, a first light source for emitting white light, made from the second light source for emitting a laser beam,
The first imaging unit images the cells illuminated by the first light source,
The imaging flow cytometer according to any one of claims 1 to 5 , wherein the second imaging unit images the cells illuminated by the second light source. The imaging flow site according to claim 6 , wherein the first light source is arranged such that the first imaging unit images a cell by transmitted illumination, and the second light source is arranged so that the second imaging unit images a cell by epi-illumination. Meter. Imaging flow cytometer according to any one of claims 1 to 7 in which the light source is a pulse light source. Control unit-out based on the average luminance of the cells of the transmitted light image, an imaging according to any one of 請 Motomeko 1-8 which Ru is displaced along a position of the sheath flow cell with the optical axis of the first imaging unit Flow cytometer. Control unit includes a displacement portion for displacing the sheath flow cell along the optical axis of the first imaging unit, and a calculation unit for calculating an average luminance of the cells of the transmitted light image, the average luminance calculated by the calculation unit becomes a predetermined value The imaging flow cytometer according to claim 9 , further comprising: a drive control unit that drives the displacement unit. The imaging flow cytometer according to claim 10, wherein the storage unit stores the average luminance calculated by the calculation unit.

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