Methods for enumerating 3-part white cell differential clusters

• US 4,596,035 A
• Filed: 06/27/1983
• Issued: 06/17/1986
• Est. Priority Date: 06/27/1983
• Status: Expired due to Term

• Current Assignee: Ortho Diagnostic Systems, Inc. (The Carlyle Group LP (Corporate Private Equity))
• Original Assignee: Ortho Diagnostic Systems, Inc. (The Carlyle Group LP (Corporate Private Equity))
• Inventors: Gershman, Russell J., Weiner, Irving L., Daniels, Arthur C.
• Primary Examiner(s): Boudreau, Leo H.
• Application Number:
  US06/508,448
• Time in Patent Office: 1,086 Days
• Field of Search: 382/6, 356/39, 356/337-340, 356/343, 377/10, 377/12, 364/416, 422/68
• US Class Current: 382/134
• CPC Class Codes: G01N 15/1456 without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
• CPC Class Codes: G01N 2021/4707 Forward scatter; Low angle scatter
• CPC Class Codes: G01N 2021/4711 Multiangle measurement

Abstract

Methods useful for differentiating and enumerating subpopulations of particles in a sample and more particularly relates to the enumeration of leukocyte subclasses. Enumeration is accomplished by utilizing two light scatter parameters and converting these parameters to a single parameter by employing a mathematical, nonlinear transformation.

Claims

1. A method for transforming forward light scatter and right angle light scatter parameter measurements into a combined one dimensional parameter for quantifying cellular subclasses of a sample comprising:

(a) passing the cells, to be differentiated into subpopulations and counted, through a zone illuminated by a focused light source, substantially one at a time;

(b) detecting light scattered by the passage of said cells through said zone in a substantially forward angle direction;

(c) further detecting light scattered by the passage of said cells through said zone at a substantially right angle direction whereby each cell may be represented by a forward angle light scatter and a right angle light scatter measurement;

(d) formulating a trajectory for connecting the cells represented by said forward angle and right angle light scatter measurements;

(e) determining a series of lines orthogonal to said trajectory describable by a nonlinear function; and

(f) counting the cells along each of said orthogonal lines whereby cellular subpopulations may be enumerated.

2. A medhod for transforming forward light scatter and right angle light scatter parameter measurements into a combined one dimensional parameter for quantifying cellular subclasses of a sample comprising:

(a) passing the cells, to be differentiated into subpopulations and counted, through a zone illuminated by a focused light source, substantially one at a time;

(b) detecting light scattered by the passage of said cells through said zone in a substantially forward angle direction;

(c) further detecting light scattered by the passage of said cells through said zone at a substantially right angle direction whereby each cell may be represented by a forward angle light scatter and a right angle light scatter measurement;

(d) formulating a trajectory for connecting the cells represented by said forward angle and right angle light scatter measurements;

(e) calculating a series of lines orthogonal to said trajectory pursuant to the nonlinear transformation;

C(x,y)=(1-mX)bY^1/2 +aX² +hX+k where X equals the log of the right angle scatter measurement, Y equals the forward angle scatter measurement, and m, b, a, h, and k are constants chosen to subtantially optimize the orthogonality of these lines to said trajectory; and

(f) counting the cells along each line of constant C whereby cellular subpopulations may be enumerated.

3. The method as provided in claim 2 wherein the sample contains leukocytes and the subpopulations to be counted comprise granulocytes, lymphocytes, and monocytes.