Apparatus for counting and determining at least one leucocytic sub-population

• US 5,138,181 A
• Filed: 10/26/1990
• Issued: 08/11/1992
• Est. Priority Date: 10/27/1989
• Status: Expired due to Term

• Current Assignee: ABX SA PARC Euromedecine
• Original Assignee: ABX
• Inventors: Lefevre, Didier, Champseix, Serge, Champseix, Henri
• Primary Examiner(s): Nelms, David C.
• Assistant Examiner(s): Messinger, Michael
• Application Number:
  US07/602,952
• Time in Patent Office: 655 Days
• Field of Search: 356/246, 356/73, 356/39, 250/573, 250/574, 250/576, 250/222.1
• US Class Current: 250/573
• CPC Class Codes: G01N 15/01 specially adapted for biological cells, e.g. blood cells investigating sedimentation of particle suspensions in blood G01N15/05
• CPC Class Codes: G01N 15/12 by observing changes in resistance or impedance across apertures when traversed by individual particles, e.g. by using the Coulter principle
• CPC Class Codes: G01N 2015/016 White blood cells
• CPC Class Codes: G01N 2015/1019 Associating Coulter-counter and optical flow cytometer [OFC]
• CPC Class Codes: G01N 2015/1024 Counting particles by non-optical means
• CPC Class Codes: G01N 2015/1413 Hydrodynamic focussing

Abstract

An apparatus is disclosed for counting and determining at least one leucocytic sub-population, comprising a case for injecting a flow of sample to be analyzed inside a measurement tank through which a light beam passes, comprising internal and external nozzles for sleeving the sample by means of a pressurized liquid. Electrodes for the resistivity measurements of the leucocytic sub-population are component elements of the injection case. Units for collecting and processing the signals delivered by the optical sensor and the electrodes are also provided.

Claims

1. Apparatus for counting and determining at least one leucocytic sub-population by resistivity based on the principle of a cell passing through an electric field where a constant current is maintained delivering a signal providing information about the volume of the cell passing through this field and using an optical method wherein a cell passes through a light ray and light absorbed by the cell is collected in an optical reader to provide information about the absorbance of the cell, said apparatus comprising a case for injecting a flow of sample to be analyzed inside a measurement tank though which passes a light beam collected by a sensor, characterized in that the sample low is sleeved hydrodynamically inside the tank by at least first and second pressurized sleeving liquids injected through the tank through at least two separate feed ducts, the second sleeving liquid flow enveloping the first sleeving liquid flow, wherein an external injection nozzle terminating in a calibrated orifice is mounted inside the case and causes an internal chamber of said nozzle to communicate with the tank, for emitting the first sleeving liquid, and wherein electrodes for measurement of the resistivity of the leucocytic sub-population are component elements of the injection case, circuits for preparing and feeding the sample flow and sleeving liquids are provided upstream of the injection case, and units for collecting and processing the signals delivered by the optical sensor and said electrodes are provided downstream of the injection case, information obtained being processed by a computer and processed information provided to a user.

2. Apparatus according to claim 1, characterized in that the information from the resistivity measurements and optical measurements is provided in a graphic representation of matrix form, for displaying the contours representative of the distribution of the populations.

3. Apparatus according to claim 1, characterized in that the calibrated orifice is offset from the light beam passing through the tank.

4. Apparatus according to claim 1, characterized in that an annular chamber is formed between a conical part of the external injection nozzle and an internal face of the case and said chamber communicates with the measurement tank and is fed with pressurized fluid through a duct for the emission of the second sleeving liquid.

5. Apparatus according to claim 1, characterized in that an internal injection nozzle is mounted inside the external nozzle and has an orifice for the injection of pressurized fluid coming from a duct which opens into a chamber inside said external nozzle in line with the calibrated orifice.

6. Apparatus according to claim 1, using a measurement tank having, on one side, a lamp emitting the light beam and, on the other side, a sensor which receives the light beam emitted by the lamp and focussed by optical elements, characterized in that the measurement tank comprises an internal passage of sufficient width that the doubly sleeved flow remains concentric and stabilized during its travel through the tank.

7. Apparatus according to claim 1, characterized in that the light beam comprises a laser ray and a sensor assembly is positioned, focussed on the measurement tank, at 90°

from a laser ray passing through it, for carrying out a cytofluorescence measurement.

8. Apparatus according to claim 1, characterized in that the resistivity measurement electrodes are formed by an end piece of the discharge duct for the sleeved flow of solution leaving the tank, forming the anode, and by the internal nozzle for injection of the solution to be analyzed, forming the cathode.

9. Apparatus according to claim 1, characterized in that the liquid to be analyzed and the sleeving fluids come from a hydraulic distribution assembly fed by a sample from a mixing and heating unit.

10. Apparatus according to claim 1, characterized in that the information from the resistivity measurements is provided in the form of a curve of distribution of volumes for localizing a main leucocytic population.

11. Apparatus according to claim 1, characterized in that the information from the optical measurements is provided by means of an absorbance distribution curve for localizing the absorbance of the leucocytes.