Flow cytometry apparatus with uniform incoherent light excitation

• US 4,600,302 A
• Filed: 03/26/1984
• Issued: 07/15/1986
• Est. Priority Date: 03/26/1984
• Status: Expired due to Fees

• Current Assignee: Becton, Dickinson & Co
• Original Assignee: Becton, Dickinson & Co
• Inventors: Sage, Burton H. Jr.
• Primary Examiner(s): Lusignan, Michael R.
• Application Number:
  US06/593,470
• Time in Patent Office: 841 Days
• Field of Search: 356/39, 356/72, 356/73, 356/317, 356/318, 362/32, 250/461.2
• US Class Current: 356/39
• CPC Class Codes: G01N 15/1434 Optical arrangements
• CPC Class Codes: G01N 15/147 the analysis being performed on a sample stream
• CPC Class Codes: G01N 2015/1497 Particle shape
• CPC Class Codes: G01N 2201/0631 Homogeneising elements

Abstract

A flow cytometry apparatus for determining one or more characteristics of particles or the like flowing in a liquid stream includes a nozzle for generating a liquid flow stream for moving particles therethrough substantially one at a time. An arc lamp provides a beam of incoherent light to illuminate the particles moving in the flow stream. A prism in the light beam modifies non-uniform incoherent light, as provided by said arc lamp, to a more uniform beam of illumination which is directed to the flowing particles. A detector is included for detecting light with respect to each moving particle and for associating the detected light with one or more characteristics of the particles.

Claims

1. A flow cytometry apparatus for determining characteristics of cells or the like flowing in a liquid stream comprising:

means for moving cells, substantially one at a time, in a liquid flow stream;

an excitation light source for providing a beam of incoherent light to illuminate said cells moving in said flow stream;

an elongate, solid, light transmissive prism having its longitudinal axis aligned substantially along said light beam between said source and said flow stream, said prism having internally reflective side surfaces and light transmissive end surfaces so that non-uniform incoherent light from said source directed into said prism through one of its end surfaces exits said prism through the opposite end surface in uniform fashion as the light is directed toward said cells;

means for detecting light associated with each moving cell as it passes through said area of illumination; and

means for using said detected light to determine one or more characteristics of said cells.

2. The apparatus of claim 1 wherein said prism is rectangular in cross-section.

3. The apparatus of claim 1 wherein said prism is positioned in said beam of light adjacent said light source.

4. The apparatus of claim 1 wherein the cross-section of said prism is substantially constant along its longitudinal axis.

5. The apparatus of claim 4 wherein the maximum cross-sectional dimension of said prism is greater than the width of the incident beam of incoherent light adapted to enter one end surface of the prism.

6. The apparatus of claim 1 wherein the end surfaces of said prism are treated with an anti-reflectance agent to improve the efficiency of light transmission therethrough.

7. The apparatus of claim 1 which further includes a lens positioned between said light source and said prism for focusing light into the prism through said one end surface thereof.

8. The apparatus of claim 7 wherein said lens is in contact with said one end surface of said prism.

9. The apparatus of claim 8 wherein said prism and said lens are integrally formed from the same material.

10. The apparatus of claim 1 wherein said prism is made of quartz.

11. The apparatus of claim 9 wherein the integrally formed prism and lens is made of quartz.

12. The apparatus of claim 1 wherein said means for detecting light includes a device for detecting fluorescence emitted by the cells passing through said area of illumination.

13. The apparatus of claim 1 wherein said means for detecting light includes a device for detecting light scattered by the cells passing through said area of illumination.

14. A flow cytometry apparatus for determining characteristics of cells or the like flowing in a liquid stream comprising:

means for moving cells, substantially one at a time, in a liquid flow stream;

an arc lamp for providing a beam of incoherent light to illuminate said cells moving in said flow stream;

an elongate, rectangular cross-section, solid, light transmissive prism having its longitudinal axis aligned substantially along said light beam and positioned between said lamp and said flow stream, said prism having internally reflective side surfaces and light transmissive end surfaces so that non-uniform light from said lamp directed to said prism through a first end surface exists said prism through the opposite, second end surface in uniform fashion toward said cells;

a lens in contact with said first end surface of said prism and positioned adjacent to and facing said lamp for focusing light into the prism through said first end surface thereof;

means for detecting light associated with each moving cell as it passes through said area of illumination; and

means for using said detected light to determine one or more characteristics of said cells.

15. The apparatus of claim 14 wherein said arc lamp is a mercury arc lamp.

16. The apparatus of claim 14 wherein the outside of said side surfaces has a protective coating thereon.