US-20260126372-A1 - FLOW CYTOMETER MODULE AND METHODS OF USE THEREOF

Abstract

A module includes one or more focusing optics, a flowcell configured to receive one or more particles, a collimated laser configured to shine a beam of light through the one or more focusing optics onto the flowcell, wherein the one or more focusing optics is configured to provide a beam of light from the collimated laser that overfills a width of the flowcell a side scatter optical train which includes one or more optics and one or more side scatter detectors, wherein the one or more side scatter detectors are configured to receive light scattered by the one or more particles, a forward scatter optical train which includes a forward scatter mask, one or more optics, and one or more forward scatter detectors, wherein the one or more forward scatter detectors are configured to receive the scattered light.

Inventors

• Robert GRANIER
• Ramin Haghgooie
• Kenneth Thomas Kotz
• Michael David SÁNCHEZ
• Daniel Morris HARTMANN
• Jesse Newton Jones, IV
• Steven M. SCHERR

Assignees

• GENERAL FLUIDICS CORPORATION

Dates

Publication Date

20260507

Application Date

20251107

Claims (20)

1. 1 . A module including: one or more focusing optics; a flowcell configured to receive one or more particles therein; a collimated laser configured to shine a beam of light through the one or more focusing optics onto the flowcell, wherein the one or more focusing optics is configured to provide a beam of light from the collimated laser that overfills a width of the flowcell; a side scatter optical train including one or more optics and one or more side scatter detectors, wherein the one or more side scatter detectors are configured to receive light scattered by the one or more particles in the flowcell; a forward scatter optical train including a forward scatter mask, one or more optics, and one or more forward scatter detectors, wherein the one or more forward scatter detectors are configured to receive the light scattered by particles flowing in the flowcell; and a processor configured to capture data from the one or more side scatter detectors and the one or more forward scatter detectors to determine at least one characteristic of the one or more particles received by the flowcell.
2. 2 . The module of claim 1 , wherein the one or more focusing optics includes a first lens and a second lens, wherein the second lens is a cylindrical lens.
3. 3 . The module of claim 1 , wherein the one or more focusing optics is configured to focus the beam from the collimated laser into an astigmatic beam onto the flowcell.
4. 4 . The module of claim 1 , wherein for the one or more focusing optics configured to provide the beam of light from the collimated laser that overfills the width of the flowcell, the beam of light overlaps one or more channel edges of the flowcell, and the beam of light has a size, uniformity, and intensity at the flowcell such that a full cross-sectional area of a channel within the flowcell is illuminated with the beam of the light that is substantially uniform in intensity.
5. 5 . The module of claim 1 , wherein the forward scatter mask further includes one or more apertures sized and positioned to produce a dual-angle differential scatter Mie map.
6. 6 . The module of claim 4 , wherein particles in a channel are not confined by sheath fluid, but are free to flow throughout the full cross-sectional area of the channel, and where furthermore, at any point in the full cross-sectional area, they will pass through a laser beam with substantially uniform intensity.
7. 7 . The module of claim 1 , wherein the one or more focusing optics is configured to provide a beam from the collimated laser on the forward scatter mask having a spatial extent such that the beam substantially avoids one or more forward scatter mask apertures.
8. 8 . The module of claim 1 , wherein the module also includes an aperture stop and the aperture stop is selected so as to limit a spatial extent of a focused beam on the forward scatter mask, and thereby substantially avoid one or more forward scatter mask apertures.
9. 9 . The module of claim 8 , wherein the aperture stop is asymmetric and is shaped so as to limit the spatial extent of the focused beam on the forward scatter mask, and thereby substantially avoid one or more forward scatter mask apertures, while maintaining as much power in the beam as possible.
10. 10 . A bidirectional cytometry sampling system including: a sample holder; a flowcell connected to the sample holder; a gantry robot including a pipettor configured to pipette a fluid sample into the sample holder; and a pump system configured to pump one or more fluids in one or more directions through the flowcell; wherein the flowcell is fluidically connected to the sample holder and the pump system.
11. 11 . The bidirectional cytometry sampling system of claim 10 , wherein the sample holder further includes: a sample holding cup; a T-junction fluidically connected to the sample holding cup; a waste channel fluidically connected to the T-junction; a valve fluidically connected to the waste channel; a port fluidically connected to the valve; an entry channel fluidically connected to the T-junction; a fluidic fitting fluidically connected to the entry channel; and a fluid conduit fluidically connected to the fluidic fitting and the flowcell.
12. 12 . The bidirectional cytometry sampling system of claim 10 , wherein the pump system further includes: a selector valve fluidically connected to a sample loop, wherein the sample loop is fluidically connected to the flowcell; a cleaning pump fluidically connected to the selector valve; and a pump fluidically connected to the selector valve and a system fluid; wherein the pump is a high-precision pump, a syringe pump, or a piston pump.
13. 13 . The bidirectional cytometry sampling system of claim 10 , wherein the bidirectional cytometry sampling system further includes: a sample loop for a sample which is fluidically connected to the flowcell and the pump system.
14. 14 . The bidirectional cytometry sampling system of claim 10 , wherein the sample holder further includes: a sample cup; a sample tube fluidically connected from the flowcell to the sample cup; and a waste port fluidically connected to the sample cup, wherein the pump system is configured to pump a fluid through the flowcell to the sample cup to clean the flowcell and the sample cup.
15. 15 . The bidirectional cytometry sampling system of claim 10 , wherein the sample holder further includes a sample cup, wherein the sample cup is configured to enable one or more samples to be loaded into the flowcell with an air gap of known size separating a sample from a fluid of a hydraulic system fluidically connected to the flowcell.
16. 16 . The bidirectional cytometry sampling system of claim 10 , wherein the sample holder further includes a sample cup, wherein the sample cup is configured to collect a used sample and a used cleaning fluid after a run of the bidirectional cytometry sampling system.
17. 17 . The bidirectional cytometry sampling system of claim 10 , wherein the sample holder further includes a sample cup, wherein the sample cup is configured to automatically direct a used sample and a used cleaning fluid to a waste and clean the sample cup.
18. 18 . A modular sheathless flow-cytometry system including: a sample holder; a pipettor configured to pipette a fluid sample into the sample holder; a pump system configured to pump one or more fluids in one or more directions through a flowcell, the flowcell being fluidically connected to the sample holder and the pump system; one or more focusing optics; a collimated laser configured to shine a beam of light through the one or more focusing optics onto the flowcell; at least one side scatter detector configured to receive light scattered by one or more particles flowing in the flowcell; at least one forward scatter detector configured to receive the light scattered by one or more particles flowing in the flowcell; a forward-scatter mask with one or more forward-scatter mask apertures to limit a collection of forward scattered light to a set of defined scatter angles; a processor configured to capture data received by the at least one side scatter detector and the data received by the at least one forward scatter detector to determine at least one characteristic of the one or more particles received by the flowcell; and a side-scatter optical path, the side-scatter optical path including: one or more filters; an automated system for inserting and removing the one or more filters; and a controller configured to operate the automated system.
19. 19 . The modular sheathless flow-cytometry system of claim 18 , wherein the flowcell further includes thicker walls configured to shift one or more contaminating particles away from a beam on the flowcell.
20. 20 . The modular sheathless flow-cytometry system of claim 18 , wherein the flowcell further includes an enclosure surrounding the flowcell configured to shift one or more contaminating particles away from a beam on the flowcell.

Description

CROSS-REFERENCE TO RELATED APPLICATION(S) This patent application claims the benefit of priority to U.S. Provisional Application No. 63/717,782, filed on Nov. 7, 2024, the entirety of which is incorporated herein by reference. TECHNICAL FIELD Various embodiments of the present disclosure relate generally to flow cytometers and, more particularly, to flow cytometer modules and methods of use. BACKGROUND Most hematology analyzers are flow cytometers that use sheath fluid to hydrodynamically focus cells in a flowcell for interrogation. Sheath fluid confines the sample of interest to a narrow stream, so that it can be interrogated with a small, focused beam of light. However, the use of sheath fluid requires additional hardware to pump the fluid through the flowcell, requires regular replacement, and greatly increases the volume of biohazardous waste that is generated. These factors serve to increase the cost-of-ownership of such systems. To address these deficiencies, some commercially available cytometers do not utilize sheath-fluid, but instead use a micro-capillary to directly pull fluid from a sample cup into an interrogation region. This simplifies the fluidics and reduces the amount of waste generated, making the system lower-cost and more portable. However, because the laser in some commercially available cytometer systems is focused down to a small spot in the flowcell, some cells in the sample may not interact with the laser beam and are therefore not counted as they flow by. As such, the precision and absolute accuracy of cell counts made with certain cytometers are typically not as good as those made by cytometers that use sheath fluid. The present disclosure is directed to overcoming one or more of these above-referenced challenges. SUMMARY OF THE DISCLOSURE Aspects of the present disclosure include a flow cytometry module and related hardware for performing hematology measurements. Aspects of the present disclosure include a modular, sheathless flow cytometer and associated hardware. In some aspects, the techniques described herein relate to a module including: one or more focusing optics; a flowcell configured to receive one or more particles therein; a collimated laser configured to shine a beam of light through the one or more focusing optics onto the flowcell; a side scatter optical train including one or more optics and one or more side scatter detectors, wherein the one or more side scatter detectors are configured to receive light scattered by the one or more particles in the flowcell; a forward scatter optical train including a forward scatter mask, one or more optics, and one or more forward scatter detectors, wherein the one or more forward scatter detectors are configured to receive the light scattered by particles flowing in the flowcell; and a processor configured to capture data from the one or more side scatter detectors and the one or more forward scatter detectors to determine at least one characteristic of the one or more particles received by the flowcell. In some aspects, the techniques described herein relate to a module, wherein the one or more focusing optics includes a first lens and a second lens, wherein the second lens is a cylindrical lens. In some aspects, the techniques described herein relate to a module, wherein the one or more focusing optics is configured to focus the beam from the collimated laser into an astigmatic beam onto the flowcell. In some aspects, the techniques described herein relate to a module, wherein the one or more focusing optics is configured to provide a beam of light from the collimated laser that overfills a width of the flowcell, overlapping one or more channel edges of the flowcell, the beam of light having a size, uniformity, and intensity at the flowcell such that a full cross-sectional area of a channel within the flowcell is illuminated with the beam of the light that is substantially uniform in intensity. In some aspects, the techniques described herein relate to a module, wherein the forward scatter mask further includes one or more apertures sized and positioned to produce a dual-angle differential scatter Mie map. In some aspects, the techniques described herein relate to a module, wherein particles in a channel are not confined by sheath fluid, but are free to flow throughout the full cross-sectional area of the channel, and where furthermore, at any point in the full cross-sectional area, they will pass through a laser beam with substantially uniform intensity. In some aspects, the techniques described herein relate to a module, wherein the one or more focusing optics is configured to provide a beam from the collimated laser on the forward scatter mask having a spatial extent such that the beam substantially avoids one or more forward scatter mask apertures. In some aspects, the techniques described herein relate to a module, wherein the module also includes an aperture stop and the aperture stop is selected so as to limit a spati