Method, system, and compositions for cell counting and analysis

• US 9,097,640 B2
• Filed: 08/20/2012
• Issued: 08/04/2015
• Est. Priority Date: 01/26/2007
• Status: Active Grant

• Current Assignee: Becton, Dickinson & Co
• Original Assignee: Becton, Dickinson & Co
• Inventors: Goldberg, Edward Michael, Brooker, Jeffrey
• Primary Examiner(s): Punnoose, Roy M
• Application Number:
  US13/590,114
• Publication Number:
  US 20130045529A1
• Time in Patent Office: 1,079 Days
• Field of Search: 356/246, 356/417, 356/418, 356/39
• US Class Current: 1/1
• CPC Class Codes: B01L 2200/0684 Venting, avoiding backpressure, avoid gas bubbles
• CPC Class Codes: B01L 2300/0822 Slides
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: G01N 15/1433 using image recognition
• CPC Class Codes: G01N 15/1434 Optical arrangements
• CPC Class Codes: G01N 15/1459 the analysis being performed on a sample stream
• CPC Class Codes: G01N 2015/1006 for cytology
• CPC Class Codes: G01N 2021/0321 One time use cells, e.g. integrally moulded
• CPC Class Codes: G01N 2021/0325 Cells for testing reactions, e.g. containing reagents
• CPC Class Codes: G01N 2021/0346 Capillary cells; Microcells
• CPC Class Codes: G01N 2021/6419 Excitation at two or more wavelengths
• CPC Class Codes: G01N 2021/6441 with two or more labels
• CPC Class Codes: G01N 21/0303 Optical path conditioning in cuvettes, e.g. windows; adapted optical elements or systems; path modifying or adjustment G01N21/031 - G01N21/15 take precedence
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6456 Spatial resolved fluorescence measurements; Imaging
• CPC Class Codes: G01N 33/49 Blood chemical methods for determining blood cell populations G01N33/5094; chemical analysis of blood groups or blood types G01N33/80
• CPC Class Codes: G01N 33/5094 for blood cell populations red blood cells G01N33/80

Abstract

The present invention provides a low cost imaged-based system for detecting, measuring and/or counting labeled features of biological samples, particularly blood specimens. In one aspect, the invention includes a system for imaging multiple features of a specimen that includes one or more light sources capable of successively generating illumination beams each having a distinct wavelength band and a plurality of differentially excitable labels capable of labeling a specimen comprising multiple features, such that each different feature is labeled with a different differentially excitable label.

Claims

1. An apparatus for analyzing in a blood specimen non-red cells labeled with a plurality differentially excitable labels, the apparatus comprising:

a sample chamber having a top wall and a bottom wall capable of containing a blood specimen, wherein the distance between the top wall and the bottom wall is configured to preclude the formation of a light-obstructing layer of red blood cells;

a blood specimen present in the sample chamber, wherein the blood specimen is labelled with a plurality of differentially excitable labels positioned in the sample chamber, such that each different non-red cell component of the blood specimen is labeled with a different differentially excitable label that emits an optical signal within the same wavelength band;

multiple light sources each capable of illuminating the blood specimen with an illumination beam having a distinct wavelength band;

a controller coupled to the multiple light sources for successively directing the illumination beam of each light source onto the specimen;

an optical system capable of collecting such emitted optical signals and forming successive images corresponding thereto on a light-responsive surface to form successive sets of image data; and

enumerating the non-red cells in the blood specimen by analyzing the successive sets of image data.

2. The system of claim 1, wherein the distance between the top surface and bottom surface of the sample chamber is equivalent to the diameter of the non-red blood cells labelled with the differentially excitable labels.

3. The system of claim 2, wherein the distance between the top wall and the bottom wall ranges from 50 μ

m to 100 μ

m.

4. An apparatus for analyzing in a blood specimen non-red cells labeled with a plurality differentially excitable labels, the apparatus comprising:

an observation area configured to receive a sample chamber having a top wall and a bottom wall capable of containing a blood specimen, wherein the distance between the top wall and the bottom wall is configured to preclude the formation of a light-obstructing layer of red blood cells;

multiple light sources each capable of illuminating the blood specimen with an illumination beam having a distinct wavelength band;

a controller coupled to the multiple light sources for successively directing the illumination beam of each light source onto the specimen;

an optical system capable of collecting such emitted optical signals and forming successive images corresponding thereto on a light-responsive surface to form successive sets of image data; and

enumerating the non-red cells in the blood specimen by analyzing the successive sets of image data.