Flow imaging cytometer

• US 5,471,294 A
• Filed: 09/29/1994
• Issued: 11/28/1995
• Est. Priority Date: 10/24/1991
• Status: Expired due to Fees

• Current Assignee: Toa Medical Electronics Co., Ltd.
• Original Assignee: Toa Medical Electronics Co., Ltd.
• Inventors: Ogino, Shinichi
• Primary Examiner(s): NOT, DEFINED
• Assistant Examiner(s): Snider, Theresa T.
• Application Number:
  US08/315,183
• Time in Patent Office: 425 Days
• Field of Search: 356/39, 356/72, 356/73, 356/318, 356/417, 250/461.2, 250/458.1, 435/287, 435/291, 435/808
• US Class Current: 356/73
• CPC Class Codes: G01N 15/1433 using image recognition
• CPC Class Codes: G01N 15/1434 Optical arrangements
• CPC Class Codes: G01N 15/1436 the optical arrangement forming an integrated apparatus with the sample container, e.g. a flow cell
• CPC Class Codes: G01N 15/1459 the analysis being performed on a sample stream
• CPC Class Codes: G01N 2015/144 Imaging characterised by its optical setup
• CPC Class Codes: G01N 2015/1488 Methods for deciding
• CPC Class Codes: G01N 2021/177 Detector of the video camera type
• CPC Class Codes: G01N 21/6456 Spatial resolved fluorescence measurements; Imaging

Abstract

Disclosed is a flow imaging cytometer having a flow cell one side of which is irradiated with excitation light, in which wavelength selecting means for reflecting excitation light and transmitting fluorescence or reflecting means for reflecting excitation light is provided on the side of the flow cell opposite the side irradiated, thereby increasing the intensity of the excitation light so that the intensity of fluorescence emitted from an irradiated cell is increased. It is possible to pick up the fluorescent images at all times. The cytometer includes a light source for emitting the excitation light, dichroic mirrors which construct the wavelength selecting means, a photomultiplier for detecting fluorescence from irradiated cells, and a cell flow-by decision circuit which, based upon a detection signal from the photomultiplier, determines whether a detected cell of interest is suited to pick-up of a fluorescent image and produces a prescribed control signal for controlling an electronic shutter.

Claims

1. A flow imaging cytometer comprising:

a flow cell through which a specimen solution containing cells to be detected is caused to flow in a state that a sheath liquid surrounds said specimen solution said cells including a cell of interest;

a light source for irradiating a flow zone of the specimen solution in said flow cell with light;

image pick-up means for picking up a still image of the cell of interest in the specimen solution; and

image processing means for executing desired data processing based upon a set of image data obtained from said image pick-up means;

wherein fluorescent images of the cells which flow through said flow cell are picked up by said image pick up means and analyzed; and

the irradiating light from said light source is light for exciting fluorescence in the cells;

the flow imaging cytometer further comprising;

reflecting means positioned at an opposite side with respect to a light entrant side of the excitation light for reflecting excitation light transmitted by said flow cell during irradiation of said specimen solution and for transmitting the fluorescence emitted by the cells in order to redirect said transmitted excitation light to said specimen solution again so as to enhance the fluorescent images of the cells by the redirected excitation light;

detecting means for detecting the fluorescence light, which is emitted by the cells in said flow cell, separately of said image pick-up means; and

decision control means for detecting a cell of interest based upon the detection signal from said detecting means, determining whether the cell of interest requires operation of said image pick-up means, and generating a prescribed control signal for pick-up of the image when the cell of interest requires pick-up of the image.

2. The flow imaging cytometer according to claim 1, wherein said reflecting means has a planar surface.

3. The flow imaging cytometer according to claim 1, wherein said reflecting means has a concave surface.

4. The flow imaging cytometer according to claim 1, wherein said reflecting means is provided in intimate contact with a surface of said flow cell.

5. The flow imaging cytometer according to claim 1, wherein said reflecting means comprises a vapor-deposited film formed on a surface of said flow cell.

6. The flow imaging cytometer according to claim 1, further comprising a second reflecting means located on the excitation light incident side of said flow cell, said second reflecting means having a concave surface provided with an aperture through which irradiating excitation light is allowed to pass.

7. A flow imaging cytometer comprising:

a flow cell through which a specimen solution containing cells to be detected is caused to flow in a state that a sheath liquid surrounds said specimen solution said cells including a cell of interest;

a light source for irradiating a flow zone of the specimen solution in said flow cell with light;

image pick-up means for picking up a still image of the cell of interest in the specimen solution; and

image processing means for executing desired data processing based upon image data from said image pick-up means;

the irradiating light from said light source is light for exciting fluorescence in the cells;

wherein fluorescent images of the cells which flow through said flow cell are picked up by said image pickup means and analyzed;

the imaging flow cytometer further comprising;

reflecting means positioned at an opposite side with respect to a light entrant side of the excitation light for reflecting excitation light transmitted by said flow cell during irradiation of said specimen solution and for transmitting the fluorescence emitted by the cells in older to redirect only said transmitted excitation light to said specimen solution again so as to enhance the fluorescent images of the cells by the redirected excitation light;

detecting means for detecting the fluorescence light, which is emitted by the cells in said flow cell, separately of said image pick-up means; and

decision control means for detecting a cell of interest based upon the detection signal from said detecting means, determining whether the cell of interest requires operation of said image pick-up means, and generating a prescribed control signal for pick-up of the image when the cell of interest requires pick-up of the image.

8. The flow imaging cytometer according to claim 7, wherein said reflecting means has a planar surface.

9. The flow imaging cytometer according to claim 7, wherein said reflecting means has a concave surface.

10. The flow imaging cytometer according to claim 7, wherein said reflecting means is provided in intimate contact with a surface of said flow cell.

11. The flow imaging cytometer according to claim 7, wherein said wavelength selecting means comprises a vapor-deposited film formed on a surface of said flow cell.

12. The flow imaging cytometer according to claim 7, wherein a second reflecting means having a concave surface provided with an aperture through which irradiating excitation light is allowed to pass is provided on the fluorescence excitation light incident side of said flow cell.