CN-121994684-A - Flow cytometry sample injection needle

Abstract

The present disclosure provides a flow cell comprising a sample injection needle for operatively connecting a sample injection line to a flow cell body. The sample injection needle includes a sample injection needle adapter having a sample tube adapter secured to the needle for transferring sample fluid from a sample injection line to a flow cell body, and a clamp for operatively connecting the sample injection needle to the flow cell body. The disclosure also provides kits comprising the sample injection needle and flow cytometry with the flow chamber, and methods of their use and assembly.

Inventors

• H. J. Rankila
• J. Manzalaga
• K. Dembsky
• C. Jiaqi

Assignees

• 贝克顿·迪金森公司

Dates

Publication Date

20260508

Application Date

20251231

Priority Date

20241106

Claims (20)

1. 1. A flow cytometer, comprising: A flow cell, comprising: A flow cell body for transporting particles in a central flow of a flow stream from a proximal end to a distal end, wherein the flow cell body comprises a flow cell conical structure at the proximal end; A sample injection needle having a passageway therethrough for conveying sample fluid from a sample injection line at a proximal end to a flow chamber body at a distal end to generate the central flow, wherein the sample injection needle comprises: a sample injection needle adapter comprising a sample tube adapter attached to a needle, and A clamp operatively connecting the sample injection needle to the flow chamber body; Wherein in a non-gripping configuration of the flow chamber, the sample injection needle adapter is free to rotate relative to the flow chamber taper and the clamp, and wherein in a gripping configuration of the flow chamber, the sample injection needle adapter is fixed relative to the flow chamber taper and the clamp; A light source configured to illuminate particles in a flow stream in a sample detection region within the flow chamber, and A detector configured to collect light emitted by the irradiated particles.
2. 2. The flow cytometer of claim 1 wherein the needle of the sample injection needle adapter comprises a proximal end attached to the sample tube adapter and a distal end positioned within the flow chamber cone structure.
3. 3. The flow cytometer of claim 2 wherein in the clamped configuration the sample injection needle adapter is secured relative to the flow cell cone structure and the clamp by compression of the clamp.
4. 4. A flow cytometer as described in claim 2 or 3 wherein the clamp is compressed by one or more fastening members.
5. 5. The flow cytometer of claim 4 wherein the clamp is compressed by a plurality of screws.
6. 6. The flow cytometer of claim 5 wherein the clamp comprises a set of holes that receive each screw of the plurality of screws.
7. 7. The flow cytometer of claim 6 wherein the flow cell body comprises a set of holes aligned with a set of clamp holes and receiving each screw of the plurality of screws.
8. 8. The flow cytometer of any of claims 5 to 7 wherein the clamp is configured such that the inclination of the sample injection needle relative to the flow cell body can be adjusted by manipulating the torque of at least one screw of the plurality of screws.
9. 9. The flow cytometer of any of claims 2 to 8 wherein the clamp comprises a distal end in contact with the sample tube adapter and a proximal end fluidly connecting a sample injection line to the sample injection needle.
10. 10. The flow cytometer of claim 9 wherein the distal end of the clamp comprises a recess within which at least a portion of the sample tube adapter is positioned and a surface that is in contact with the proximal end of the flow cell body.
11. 11. The flow cytometer of claim 9 or 10 wherein the proximal end of the clamp comprises a connector configured to minimize dead volume of sample fluid as it flows from the sample injection line to the sample injection needle.
12. 12. The flow cytometer of any of claims 2 to 11 wherein the sample tube adapter comprises a proximal end positioned in a recess of the clamp and a distal end in contact with the proximal end of the flow cell body.
13. 13. The flow cytometer of claim 11 or 12 wherein the sample tube adapter comprises a flange in contact with the proximal end of the flow cell body.
14. 14. The flow cytometer of any of claims 11 to 13 wherein the distal end of the sample tube adapter is pressed against the proximal end of the flow cell body by the clamp such that the distal end of the needle of the sample injection needle adapter is in a fixed position within the flow cell cone structure.
15. 15. The flow cytometer of any of claims 2 to 14 wherein the needle of the sample injection needle adapter tapers at the distal end.
16. 16. The flow cytometer of any of claims 2 to 15 wherein the distal end of the needle of the sample injection needle adapter is positioned within the flow chamber cone structure such that a complete central flow can be maintained when flow conditions change by an order of magnitude or more.
17. 17. The flow cytometer of any of claims 2 to 16 wherein the flow cell body comprises a sheath fluid introduction port for delivering sheath fluid to the flow cell cone structure.
18. 18. The flow cytometer of claim 17 wherein the distal end of the needle of the sample injection needle adapter is separated from the sheath fluid introduction port by a longitudinal distance in the range of 17mm to 26 mm.
19. 19. The flow cytometer of any of claims 2 to 18 wherein the distal end of the flow cell body comprises a cuvette for transporting particles in a central stream through the sample detection zone, wherein the cuvette is positioned at the distal end of the flow cell body by a clamp secured to the flow cell body.
20. 20. The flow cytometer of claim 19 wherein the cuvette is positioned by the flow cell body clamp such that the sample detection region is optimally aligned with the cuvette for optical detection of particles in the central stream.

Description

Flow cytometry sample injection needle Technical Field Characterization of analytes in biological fluids has become an important component of biological research, medical diagnosis, and assessment of overall patient health and wellbeing. Detection of analytes in biological fluids, such as human blood or blood-derived products, may provide results that may be useful in determining treatment regimens for patients suffering from a variety of disease states. Background Flow cytometry is a technique for characterizing and often sorting biological materials, such as cells in a blood sample or other types of biological or chemical samples, of particles of interest. Flow cytometers typically include a sample reservoir for receiving a fluid sample, such as a blood sample, and a sheath fluid reservoir containing a sheath fluid. The flow cytometer delivers particles (including cells) in the fluid sample as a stream of cells to the flow cell while also directing sheath fluid to the flow cell. To characterize the composition of the flow stream, the flow stream is irradiated with light. Changes in the material in the flow stream (e.g., morphology or the presence of fluorescent markers) may result in changes in the observed light, and these changes can be used for characterization and separation. In order to characterize the components in the flow stream, light must be directed onto the flow stream and collected. The light source of the flow cytometer may vary and may include one or more of a broad spectrum lamp, a light emitting diode, and a single wavelength laser. The light source is aligned with the flow stream and the optical response from the illuminated particles is collected and quantified. The separation of biological particles can be achieved by adding a sorting or collecting function to the flow cytometer. In the separated stream, particles detected to have one or more desired characteristics are separated from the sample stream by mechanical or electrical removal. One common flow sorting technique employs droplet sorting, in which a stream containing linearly separated particles is broken up into droplets. Droplets containing the particles of interest are charged and deflected into a collection tube when passing through an electric field. Typically, particles that are linearly separated in the stream are characterized as they pass through a viewpoint located directly below the tip of the nozzle. Once a particle is identified that meets one or more desired criteria, its time to reach the point of droplet break and separate into droplets from the stream can be predicted. Ideally, the convection current is briefly charged immediately before the drop containing the selected particle breaks away from the fluid stream, and then grounded immediately after the drop breaks away. The drop to be sorted remains charged while it is detached from the fluid flow, while all other drops remain uncharged. Some flow cell systems employ pressure-driven fluidic techniques to simultaneously deliver sample fluid and sheath fluid to a flow cell. In these systems, the sample fluid and sheath fluid are delivered to a flow chamber containing a detection zone (i.e., the zone where the particles are illuminated by a light source) at a pressure above ambient. By varying the pressure in the sample tube and/or sheath fluid reservoir that is sent to the flow cell, a change in the flow rate through the flow cell of the pressure-driven fluidic system can be achieved. The ratio of sample fluid and sheath fluid flowing through the flow chamber is controlled by both the pressure levels in the sample tube and sheath fluid reservoir and the ratio of the resistances of the sample fluid path and sheath fluid path. Alternatively, the flow cell system is implemented using vacuum driven fluidic techniques, wherein a vacuum pump draws a vacuum downstream of the flow chamber, and the sample fluid and sheath fluid are maintained at ambient pressure. For a vacuum driven fluidic system, by varying the vacuum level drawn by the vacuum pump, a change in the flow rate through the flow chamber can be achieved, and the ratio of sample fluid to sheath fluid flowing through the flow chamber is controlled by the ratio of the resistances of the sample fluid path and the sheath fluid path. To achieve single particle level characterization and separation of biological materials, some flow cytometers are equipped with an injection needle for introducing a sample fluid into the flow cell. With the sample injection needle, the sample fluid may be mixed with the sheath fluid in the flow chamber in a form sufficient to form a focused particle core stream containing the sample fluid. The central stream may then convey the particles in a single train through a detection zone and/or sorting device. This technique is known as hydrodynamic focusing. Disclosure of Invention The present inventors have recognized that imaging particles using a flow cytometer requires precise control