Detection and fluidic system of a flow cytometer

• US 8,303,894 B2
• Filed: 08/16/2010
• Issued: 11/06/2012
• Est. Priority Date: 10/13/2005
• Status: Active Grant

• Current Assignee: Accuri Cytometers, Inc. (Becton Dickinson & Co)
• Original Assignee: Accuri Cytometers, Inc. (Becton Dickinson & Co)
• Inventors: Rich, Collin A.
• Primary Examiner(s): Turk, Neil N
• Application Number:
  US12/857,290
• Publication Number:
  US 20100319469A1
• Time in Patent Office: 813 Days
• Field of Search: None
• US Class Current: 422/67
• CPC Class Codes: G01N 1/14 Suction devices, e.g. pumps; Ejector devices
• CPC Class Codes: G01N 15/06 Investigating concentration of particle suspensions by weighing G01N5/00; investigating sedimentation of particle suspensions G01N15/04; investigating individual particles G01N15/10
• CPC Class Codes: G01N 15/1404 Handling flow, e.g. hydrodynamic focusing
• CPC Class Codes: G01N 2015/1413 Hydrodynamic focussing
• CPC Class Codes: G05D 7/0682 using a plurality of flow sources
• CPC Class Codes: Y10T 137/0402 Cleaning, repairing, or assembling
• CPC Class Codes: Y10T 137/85978 With pump
• CPC Class Codes: Y10T 137/85986 Pumped fluid control
• CPC Class Codes: Y10T 436/115831 Condition or time responsive
• CPC Class Codes: Y10T 436/117497 with a continuously flowing sample or carrier stream

Abstract

The fluidic system including a sheath pump that pumps sheath fluid from a sheath container into an interrogation zone, a waste pump that pumps waste fluid from the interrogation zone to a waste container, in which the flow rate of the sheath fluid is different from the flow rate of the waste fluid thereby drawing a sample fluid from a sample container into the interrogation zone, a detection system that provides a data set of input signals from the sample fluid, an analysis engine that recognizes aggregate particle events in the data set, and a controller that automatically adjusts the flow rate of the sample fluid into the interrogation zone based on the recognition of aggregate particle events, by controlling at least one of the flow rates of the sheath fluid and the waste fluid.

Claims

1. A fluidic system for drawing sample fluid from a sample container, into an interrogation zone of a flow cytometer, comprising:

a sheath pump that pumps sheath fluid at a sheath flow rate from a sheath container through a flow cell into the interrogation zone of the flow cytometer, wherein the flow cell is fluidically coupled to the sample container;

a waste pump that pumps waste fluid at a waste flow rate from the interrogation zone into a waste container;

wherein the sheath flow rate is different from the waste flow rate, thereby creating a pressure differential, and

wherein the pressure differential draws the sample fluid from the sample container, through the flow cell with the sheath fluid into the interrogation zone at a sample flow rate;

a detection system coupled to the interrogation zone that provides a data set of input signals from the sample fluid;

an analysis engine that recognizes aggregate particle events in the data set; and

a controller that automatically adjusts the sample flow rate based on the recognition of aggregate particle events by controlling at least one of the sheath flow rate and the waste flow rate to adjust the pressure differential.

2. The fluidic system of claim 1, wherein the controller automatically adjusts a core stream diameter of the sample fluid in response to the recognition of an aggregate particle event.

3. The fluidic system of claim 2, wherein the controller decreases the core stream diameter when the analysis engine recognizes an aggregate particle event in the data set.

4. The fluidic system of claim 3, wherein the controller decreases the core stream diameter to approximately the diameter of a single sample particle in response to the recognition of an aggregate particle event, such that sample particles in the sample fluid flow in single file into the interrogation zone.

5. The fluidic system of claim 3, wherein the analysis engine further recognizes non-aggregate particles events, wherein the controller increases the core stream diameter when the analysis engine recognizes a non-aggregate particle event in the data set.

6. The fluidic system of claim 2, wherein the controller increases the core stream diameter when the analysis engine recognizes an aggregate particle event in the data set.

7. The fluidic system of claim 1, wherein the controller automatically adjusts the sample flow rate after a predetermined number of aggregate particle events is recognized.

8. The fluidic system of claim 1, wherein the analysis engine further recognizes non-aggregate particle events, wherein the controller automatically adjusts the sample flow rate after a predetermined ratio of aggregate particle events to non-aggregate particle events is recognized.

9. The fluidic system of claim 1, wherein the analysis engine recognizes a “

peak-trough-peak”

waveform produced by aggregate particle events.

10. The fluidic system of claim 9, wherein the detection system includes a detector that receives photonic inputs from the interrogation zone and has a wide dynamic range defined as greater than or equal to 100 dB.

11. A method for drawing sample fluid from a sample container into an interrogation zone of a flow cytometer, comprising:

simultaneously pumping sheath fluid at a sheath flow rate from a sheath container through a flow cell, fluidicially coupled to the sample container, into the interrogation zone of the flow cytometer and pumping waste fluid at a waste flow rate from the interrogation zone into a waste container,

wherein the sheath flow rate is different from the waste flow rate, thereby creating a pressure differential, and

wherein the pressure differential draws the sample fluid from the sample container, through the flow cell with the sheath fluid into the interrogation zone at a sample flow rate;

collecting a data set of input signals from the sample fluid;

recognizing aggregate particle events in the data set with use of an algorithm; and

automatically adjusting the sample flow rate based on the recognition of aggregate particle events in the data set, wherein adjusting the sample flow rate includes controlling at least one of the sheath flow rate and waste flow rate to adjust the pressure differential.

12. The method of claim 11, wherein adjusting the sample flow rate further includes adjusting a core stream diameter of the sample fluid.

13. The method of claim 12, wherein adjusting the core stream diameter of the sample fluid includes decreasing the core stream diameter after recognizing an aggregate particle event in the data set.

14. The method of claim 13, wherein decreasing the core stream diameter includes decreasing the core stream diameter to a diameter that forces particles in the sample fluid to flow in single file into the interrogation zone.

15. The method of claim 13, further comprising recognizing non-aggregate particle events in the data set, wherein adjusting the core stream diameter of the sample fluid further includes increasing the core stream diameter after recognizing a non-aggregate particle event in the data set.

16. The method of claim 15, further comprising repeating the step of adjusting the sample flow rate after at least one of (a) recognizing aggregate particle events and (b) recognizing non-aggregate particle events.

17. The method of claim 15, wherein adjusting the sample flow rate includes adjusting the sample flow rate after a predetermined ratio of aggregate particle events to non-aggregate particle events is recognized.

18. The method of claim 12, wherein adjusting the core stream diameter of the sample fluid includes increasing the core stream diameter in response to recognition of an aggregate particle event in the data set.

19. The method of claim 12, wherein adjusting the flow rate of the sample fluid includes adjusting the flow rate of the sample fluid after a predetermined number of aggregate particle events is recognized.

20. The method of claim 12, wherein collecting a data set of input signals from the sample fluid includes detecting a wide dynamic range of inputs greater than or equal to 100 dB.