Flow cytometer apparatus and method

• US 8,664,006 B2
• Filed: 03/01/2013
• Issued: 03/04/2014
• Est. Priority Date: 03/28/2003
• Status: Active Grant

• Current Assignee: Inguran LLC
• Original Assignee: Inguran LLC
• Inventors: Durack, Gary, Wallace, Jeffrey D., Vandre, Gary P., Westfall, Lon A., Hatcher, Jeremy T., Nayak, Niraj V.
• Primary Examiner(s): Wallenhorst, Maureen
• Application Number:
  US13/782,417
• Publication Number:
  US 20130224786A1
• Time in Patent Office: 368 Days
• Field of Search: 422/67, 422/68.1, 422/73, 422/82.05, 422/501, 422/509, 422/515, 436/63, 436/164, 436/165, 436/43, 436/50, 436/55, 436/180, 435/2, 435/287.1, 435/286.4, 435/288.7, 435/366, 435/29
• US Class Current: 436/180
• CPC Class Codes: A01N 1/0284 Temperature processes, i.e. using a designated change in temperature over time
• CPC Class Codes: C12N 5/06 Animal cells or tissues; Human cells or tissues preservation of living cells or tissues A01N1/02
• CPC Class Codes: C12N 5/061 Sperm cells, spermatogonia
• CPC Class Codes: C12N 5/0612 sorting of gametes, e.g. according to sex or motility
• CPC Class Codes: C12Q 1/02 involving viable microorganisms
• CPC Class Codes: C12Q 3/00 Condition responsive control processes apparatus therefor C12M1/36; controlling or regulating in general G05
• 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 15/1404 Handling flow, e.g. hydrodynamic focusing
• CPC Class Codes: G01N 15/1459 the analysis being performed on a sample stream
• CPC Class Codes: G01N 15/1468 with spatial resolution of the texture or inner structure of the particle
• CPC Class Codes: G01N 15/147 the analysis being performed on a sample stream
• CPC Class Codes: G01N 15/149 specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: G01N 2015/1006 for cytology
• CPC Class Codes: G01N 2015/1406 Control of droplet point
• CPC Class Codes: G01N 2015/1415 Control of particle position
• CPC Class Codes: G01N 2021/6439 with indicators, stains, dyes, tags, labels, marks
• CPC Class Codes: G01N 21/63 optically excited
• 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 33/48 Biological material, e.g. blood, urine G01N33/02, G01N33/26, G01N33/44, G01N33/46 take precedence; Haemocytometers counting blood corpuscules distributed over a surface by scanning the surface G06M11/02
• CPC Class Codes: G01N 33/5005 involving human or animal cells immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025
• CPC Class Codes: Y10T 436/10 : Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
• CPC Class Codes: Y10T 436/115831 : Condition or time responsive
• CPC Class Codes: Y10T 436/12 : Condition responsive control
• CPC Class Codes: Y10T 436/25 : including sample preparation
• CPC Class Codes: Y10T 436/2525 : Stabilizing or preserving
• CPC Class Codes: Y10T 436/25375 : Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
• CPC Class Codes: Y10T 436/255 : including use of a solid sorbent, semipermeable membrane, or liquid extraction
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

A nozzle system for use in a flow cytometer for analyzing particles in a fluid stream and a method of orienting particles in a flow cytometer. The nozzle system may include a nozzle having a longitudinal axis, an interior surface defining a fluid flow path for orienting particles in the nozzle, and a downstream orifice through which a fluid stream exits. The nozzle system may also include a conduit positioned to introduce a core stream containing particles at a location offset from the longitudinal axis and a baffle downstream of the conduit to deflect the fluid stream. The method may include the steps of causing a sheath fluid to flow through the nozzle, introducing a core fluid stream into the sheath fluid at a location offset to the longitudinal axis of the nozzle, and deflecting the core fluid stream with a baffle.

Claims

1. A nozzle system for use in a flow cytometer for analyzing particles in a fluid stream, said fluid stream comprising a sheath stream surrounding a core stream containing said particles, said nozzle system comprising:

a nozzle having a longitudinal axis, an interior surface defining a flow path for said fluid stream, and an orifice at a downstream end of the nozzle for exit of the fluid stream from the nozzle, said interior surface being shaped to define at least one torsional zone comprising an axially tapered region having a generally elliptical cross section for orienting said particles in a desired orientation as the fluid stream flows through the torsional zone toward the orifice;

a conduit from a particle source to the nozzle, said conduit being positioned to introduce said core stream into the nozzle at a location that is offset with respect to the longitudinal axis of the nozzle; and

a baffle located downstream of the conduit and upstream of the orifice to deflect the fluid stream as is moves along the flow path.

2. The nozzle system of claim 1, wherein said location is upstream from the at least one torsional zone.

3. The nozzle system of claim 1, wherein the at least one torsional zone comprises a first torsional zone, the nozzle further comprising a second torsional zone.

4. The nozzle system of claim 3, wherein said location is upstream from the first torsional zone and at least a portion of the second torsional zone.

5. The nozzle system of claim 3, wherein the second torsional zone comprises an axially tapered region of the nozzle having a generally elliptical cross section.

6. The nozzle system of claim 5, wherein a major axis of the generally elliptical cross section of the first torsional zone is oriented in a different direction than the major axis of the generally elliptical cross section of the second torsional zone.

7. The nozzle system of claim 1, wherein the at least one torsional zone comprises a first torsional zone, and a second torsional zone.

8. The nozzle system of claim 7, wherein the first torsional zone comprises a first axially tapered region having a first angle of taper and a second axially tapered region having a second angle of taper.

9. The nozzle system of claim 8, wherein the second torsional zone comprises a third axially tapered region having a third angle of taper.

10. The nozzle system of claim 9, wherein interior surface of the nozzle includes a fourth non-tapered region coaxial with and downstream from the first, second and third axially tapered regions, the fourth non-tapered region being adjacent to the third axially tapered region.

11. A method of orienting particles in a flow cytometer comprising:

causing a sheath fluid to flow through a nozzle having a longitudinal axis and at least one torsional zone comprising an axially tapered region of the nozzle having a generally elliptical cross section;

introducing a core fluid stream containing particles into the sheath fluid stream at a location that is offset from said longitudinal axis for flow of the sheath fluid stream and core fluid stream through the at least one torsional zone;

deflecting the core fluid stream with a baffle as the core fluid stream flows through the nozzle causing the core stream to enter the at least one torsional zone in a manner offset from the longitudinal axis; and

subjecting particles within the core stream to hydrodynamic forces that tend to cause the particles to assume a desired orientation.

12. The method of claim 11, further comprising the step of causing the core stream to flow along a flow path, a portion of said flow path being offset from the longitudinal axis of the nozzle, and subjecting said particles to hydrodynamic orientation forces generated by said torsional zone while said particles are moving along said offset portion of the flow path.

13. The method of claim 11, wherein said at least one torsional zone constitutes a first torsional zone, the method further comprising the step of causing the sheath fluid stream and core fluid stream to flow through a second torsional zone for orienting the particles in a desired orientation.

14. The method of claim 13, wherein said second torsional zone comprises a tapered region in the nozzle having generally elliptical cross sectional area.

15. The method of claim 14, wherein a major axis of the generally elliptical cross sectional area of the first torsional zone is oriented in a different direction than the major axis of the generally elliptical cross sectional area of the second torsional zone.

16. The method of claim 14, wherein a major axis of the generally elliptical cross sectional area of the first torsional zone is perpendicular to the major axis of the generally elliptical cross sectional area of the second torsional zone.