Flow cytometry

• US 4,818,103 A
• Filed: 01/20/1987
• Issued: 04/04/1989
• Est. Priority Date: 05/15/1981
• Status: Expired due to Term

• Current Assignee: Ratcom
• Original Assignee: Ratcom
• Inventors: Thomas, Richard A., Thornthwaite, Jerry T., Eggelston, Ross W.
• Primary Examiner(s): McGraw, Vincent P.
• Application Number:
  US07/005,280
• Time in Patent Office: 805 Days
• Field of Search: 356/39, 356/72, 356/73, 356/246, 648/356, 324/71.4
• US Class Current: 356/72
• CPC Class Codes: G01N 15/13 Details pertaining to apertures
• CPC Class Codes: G01N 15/1404 Handling flow, e.g. hydrodynamic focusing
• CPC Class Codes: G01N 2015/1019 Associating Coulter-counter and optical flow cytometer [OFC]
• CPC Class Codes: G01N 2015/1415 Control of particle position
• CPC Class Codes: G01N 2015/1477 Multiparameters

Abstract

A flow cytometer has an aperture in which an inlet chamber and/or an outlet chamber have predetermined geometric relationships with respect to the aperture for decreasing electronic edge effects and increasing the sensitivity of electronic particle volume measurements. In one embodiment, the aperture is triangular and is formed by assembling a plurality of truncated pyramids with their truncated surfaces defining aperture walls. At least one of the elements forming the aperture walls is either part of a lens systems or a transparent cover plate so that simultaneous optical and electronic cell volume measurements can be performed. In one embodiment, a piezoelectric transducer is provided to sonically clear any aperture clogging.

Claims

1. A flow transducer comprising means defining an aperture having an axis, said aperture having at least one flat side, means defining an inlet chamber and an outlet chamber immediately adjacent the aperture along its axis, at least one of said inlet and outlet chamber having walls disposed at an angle of at least 5°

relative to the plane of the aperture, said at least one of said inlet and outlet chambers at a distance from the aperture of twice the width of the aperture in a plane through its axis having a cross-section area at least 10 times the cross-sectional area of said aperture.

2. A transducer for a flow cytometer in accordance with claim 1 wherein said transducer is formed of a plurality of solid polygons joined such that adjacent flat surfaces of said polygons define walls of the aperture for the flow cytometer, and others of the surfaces of said polygons define said at least one of said inlet and outlet chambers.

3. A flow cytometer for making simultaneous electronic and optical measurements on a particle flowing through a sensing zone thereof, comprising a transducer formed of a plurality of solid polygons joined such that adjacent flat surfaces of said polygons define a polygonal aperture at the sensing zone and other surfaces of said polygons define inlet and outlet chambers to said aperture, at least one of said inlet and outlet chambers having a predefined geometric relation to said aperture, said inlet and outlet chambers defining an arch shaped fluid passageway with the aperture at the arch vertex, means for establishing a flow of electrolyte through said aperture, said inlet and outlet chambers being configured to establish laminar flow of said electrolyte through said aperture, injector means for injecting samples of particles into said laminar flow of electrolyte, one or more electrodes coupled to said inlet chamber and one or more electrodes coupled to said outlet chamber, means for establishing a current flow through said aperture between said electrodes, monitoring means for monitoring the electrical current flow through said aperture, at least one of the solid polygons defining said aperture being an element in an optical measurement system, said system including means for introducing exciting light through said at least one solid polygon into said aperture, and means for collecting light from a sample particle in said aperture.

4. A flow cytometer in accordance with claim 3 wherein the predetermined geometric relation between said at least one of the inlet and outlet chambers and the aperture is such that walls of said at least one of the inlet and outlet chambers join the aperture at an angle equal to at least 5°

with respect to the plane of the aperture, and in which at a distance in the said at least one of inlet and outlet chambers from the aperture equal to twice the width of the aperture along any plane through its axis, the cross-sectional area of said at least one of the inlet and outlet chambers being at least ten times the cross-sectional area of the aperture.

5. A flow cytometer for making simultaneous electronic and optical measurements on a particle flowing through a sensing zone thereof, comprising a transducer formed of a plurality of solid polygons joined such that adjacent surfaces of said polygons define an aperture at the sensing zone and other surfaces of said polygons define inlet and outlet chambers to said aperture at least one of said inlet and outlet chambers having a predefined geometric relation to said aperture, said inlet and outlet chambers defining an arch shaped fluid passageway with the aperture at the arch vertex, means for establishing a flow of electrolyte through said aperture, said inlet and outlet chambers being configured to establish laminar flow of said electrolyte through said aperture, injector means for injecting samples of particles into said laminar flow of electrolyte, one or more electrodes coupled to said inlet chamber and one or more electrodes coupled to said outlet chamber, means for establishing a current flow through said aperture between said electrodes, monitoring means for monitoring the electrical current flow through said aperture, at least one of the solid polygons defining said aperture being an element in an optical measurement system, said system including means for introducing exciting light through at least one solid polygon into said aperture, and means for collecting light from a sample particle in said aperture.

6. A flow transducer comprising means defining an aperture having an axis, means defining an inlet chamber and an outlet chamber immediately adjacent the aperture along its axis at least one of said inlet and outlet chambers having walls disposed at an angle of at least 5°

relative to the plane of the aperture, said at least one of said inlet and outlet chambers at a distance from the aperture of twice the width of the aperture in a plane through its axis having a cross-sectional area which is rotationally asymmetric with respect to the axis of the aperture and which is at least 10 times the cross-sectional area of said aperture.