Optical particle analyzing apparatus having two types of light source

• US 5,260,764 A
• Filed: 05/29/1990
• Issued: 11/09/1993
• Est. Priority Date: 02/08/1990
• Status: Expired due to Fees

• Current Assignee: Toa Medical Electronics Co., Ltd.
• Original Assignee: Toa Medical Electronics Co., Ltd.
• Inventors: Seshimo, Hiroyuki, Tohori, Hidemichi, Fukuda, Masakazu, Nakamoto, Hiroyuki
• Primary Examiner(s): McGraw, Vincent P.
• Assistant Examiner(s): HANTIS, KAREN
• Application Number:
  US07/530,102
• Time in Patent Office: 1,260 Days
• Field of Search: 356/73, 356/39, 356/72, 356/317, 356/318, 356/417, 356/336, 250/458.1, 250/459.1, 250/461.1, 250/461.2
• US Class Current: 356/73
• CPC Class Codes: G01N 15/14 Optical investigation techniques, e.g. flow cytometry
• CPC Class Codes: G01N 15/1459 the analysis being performed on a sample stream
• CPC Class Codes: G01N 2015/1477 Multiparameters
• CPC Class Codes: G01N 2021/1791 stroboscopic; pulse gated; time range gated
• CPC Class Codes: G01N 2021/6439 with indicators, stains, dyes, tags, labels, marks
• CPC Class Codes: G01N 21/47 Scattering, i.e. diffuse reflection G01N21/25, G01N21/41 take precedence G01N21/55 takes precedence

Abstract

An optical particle analyzing apparatus includes a flow cytometer in which laser light and lamp light having a wavelength shorter than that of the laser light are used simultaneously when particles in a blood specimen or the like are analyzed by the flow cytometer. Scattered light emitted by a particle due to irradiation with the laser light and fluorescence emitted by the particle due to irradiation with the lamp light are detected. A signal indicative of the scattered light is used as a timing signal to detect the fluorescence. This arrangement makes it possible to reduce the size and cost of the apparatus.

Claims

1. An optical particle analyzing apparatus for classifying and enumerating particles, comprising:

a flow cell;

means for passing a fine stream of cells stained with a fluorescent stain through said flow cell;

means for continuously irradiating an irradiating zone in the fine stream of cells in said flow cell with two kinds of light;

means for detecting respective ones of forward scattered light and fluorescence produced by the cells in the irradiating zone;

means for processing and analyzing signals outputted by said detecting means, wherein said two kinds of light which irradiate the irradiating zone are a laser light beam emitted by a semiconductor laser light source and a lamp light beam, which has a wavelength shorter than that of the laser light beam, emitted by a lamp light source, the laser light beam and lamp light beam intersect each other at the irradiating zone to irradiate the same, and the irradiating laser light beam is narrowed to a beam narrower than the lamp light beam in a direction in which the cells flow; and

a signal processor for signal-processing a forward scattered-light signal which results from the laser light beam and is detected in the irradiating zone, and a fluorescence signal which results from the lamp light beam and is detected in the irradiating zone, wherein when the fluorescence signal is signal-processed by said signal processor, the forward scattered-light signal is used as a timing signal for this signal processing so that the value of the fluorescence signal at the moment the scattered-light signal peaks is taken as the intensity of fluorescence.

2. The apparatus according to claim 1, wherein the irradiating light beams composed of the laser light beam and lamp light beam intersect each other at an angle of approximately 90°

, and forward-scattered light and side fluorescence which result from the laser light beam and lamp light beam, respectively, and are emitted in the same direction, are detected upon undergoing wavelength selection.

3. The apparatus according to claim 1, wherein the irradiating light beams composed of the laser light beam and lamp light beam intersect each other at an angle of approximately 90°

, and forward-scattered light which results from the laser light beam and is emitted in a first direction, as well as side-scattered light and back fluorescence which result from the laser light beam and lamp light beam, respectively, and are emitted in a second direction, is detected.

4. An optical analyzer for classifying and enumerating cells, comprising:

a flow cell having an irradiating zone through which a stream of cells stained with a fluorescent stain flows;

a semiconducting laser for generating a laser beam which irradiates said stream of cells at said irradiating zone of said flow cell;

an optical energy source for generating a light beam having a wavelength shorter than the wavelength of said laser beam which continuously irradiates said stream of cells at said irradiating zone of said flow cell and intersects said laser beam at said irradiating zone of said flow cell, whereby said stained cells fluoresce when irradiated by said light beam;

a condensing lens system receiving said laser beam for narrowing said laser beam narrower than said light beam in the direction in which said stream of cells flow;

a signal processor which detects the interaction of said laser and light beams with said stream of cells for generating a first signal corresponding to the irradiation of said stream of cells by said laser and a second signal corresponding to the fluorescence resulting from the irradiation of said stream of cells by said light beam; and

a data processor coupled to said signal processor for processing and analyzing said second signal generated at the same time said first signal is generated.

5. The optical analyzer of claim 4 wherein said laser and light beams intersect at an angle of approximately ninety degrees.

6. The optical analyzer of claim 4 wherein

said irradiation of said stream of cells by said laser beam produces forward scattered light;

said irradiation of said stream of cells by said light source produces side fluorescence emitted in the same direction as said forward scattered light; and

said signal processor uses wavelength detection to differentiate between said forward scattered light and said side fluorescence.

7. An optical analyzer for classifying and enumerating cells, comprising:

a flow cell having an irradiating zone through which a stream of cells stained with a fluorescent stain flows;

a semiconducting laser for generating a laser beam which irradiates said stream of cells at said irradiating zone of said flow cell to produce forward scattered light which is emitted in a first direction and to produce side scattered light which is emitted in a second direction;

an optical energy source for generating a light beam having a wavelength shorter than the wavelength of said laser beam which continuously irradiates said stream of cells at said irradiating zone of said flow cell and intersects said laser beam at said irradiating zone of said flow cell to produce back fluorescence emitted in said second direction;

a condensing lens system receiving said laser beam for narrowing said laser beam narrower than said light beam in the direction in which said stream of cells flow;

a signal processor which detects said forward scattered light, side scattered light, and back fluorescence and generates a first signal corresponding to said forward scattered light, a second signal corresponding to said side scattered light, and a third signal corresponding to said back fluorescence;

a data processor coupled to said signal processor for processing and analyzing said second and third signals generated at the same time said first signal is generated.

8. The optical analyzer of claim 7 wherein said signal processor uses wavelength detection to differentiate between said forward scattered light, side scattered light, and back fluorescence.