Automated set-up for cell sorting
Apparatus and methods are described for automatically performing set-up steps for flow cytometry operations. The invention provides for the spatial determination of a flow stream and the subsequent automatic alignment of analysis devices and/or collection vessels. The automatic determination of flow stream properties provides for the automatic configuration flow cytometer parameters.
1. A method for adjusting one or more parameters of a flow cytometer, the method comprising:
injecting a sample into a flow cell of a flow cytometer, wherein the flow cytometer comprises a system comprising a processor with memory operably coupled to the processor wherein the system is automated to:
capture one or more images of the flow cytometer flow stream comprising the sample in a detection field;
determine the width of the flow stream in the detection field;
generate a data signal corresponding to a ratio of the determined width of the flow stream and a width of the flow stream that is expected based on nozzle orifice size of the flow cell; and
adjust one or more parameters of the flow cytometer in response to the data signal.
2. The method according to claim 1 , wherein the one or more parameters of the flow cytometer is selected from the group consisting of hydrostatic pressure, sheath fluid pressure, flow stream charge, deflection voltage, drop drive frequency, charge correction value, drop delay, drop amplitude and charge phase.
3. The method according to claim 2 , wherein the processor is configured to adjust sheath fluid pressure based on the determined flow cell nozzle orifice diameter.
4. The method according to claim 2 , wherein the processor is configured to adjust drop drive frequency based on the determined flow cell nozzle orifice diameter.
5. The method according to claim 1 , wherein the processor is configured to determine the nozzle orifice diameter of the flow cell and generate a data signal corresponding to the flow cell nozzle orifice diameter based on the determined width of the flow stream.
6. The method according to claim 5 , wherein the processor is configured to adjust one or more parameters of the flow cytometer based on the determined flow cell nozzle orifice diameter.
7. The method according to claim 1 , wherein the images of the flow cytometer flow stream are captured with a charge-coupled device (CCD) camera.
8. The method according to claim 1 , wherein the width of the flow stream is determined from the images based on droplet volume.
9. The method according to claim 1 , wherein the images of the flow stream are captured at the break-off point of the flow stream.
10. A flow cytometer comprising:
a flow cell configured to propagate a sample in a flow stream;
a sensor configured to capture one or more images of the flow stream;
a processor comprising memory operably coupled to the processor, wherein the memory includes instructions stored thereon to determine the width of the flow stream from the one or more images, generate a data signal corresponding to a ratio of the determined width of the flow stream and a width of the flow stream that is expected based on nozzle orifice size of the flow cell, and automatically adjust one or more parameters of the flow cytometer in response to the data signal.
11. The flow cytometer according to claim 10 , wherein the parameters of the flow cytometer is selected from the group consisting of hydrostatic pressure, sheath fluid pressure, flow stream charge, deflection voltage, drop drive frequency, charge correction value, drop delay, drop amplitude and charge phase.
12. The flow cytometer according to claim 10 , wherein the processor is configured to automatically adjust sheath fluid pressure based on the determined flow cell nozzle orifice diameter.
13. The flow cytometer according to claim 10 , wherein the processor is configured to automatically adjust drop drive frequency based on the determined flow cell nozzle orifice diameter.
14. The flow cytometer according to claim 10 , wherein the processor is configured to determine the nozzle orifice diameter of the flow cell and generate a data signal corresponding to the flow cell nozzle orifice diameter based on the determined width of the flow stream.
15. The flow cytometer according to claim 14 , wherein the processor is configured to automatically adjust one or more parameters of the flow cytometer based on the determined flow cell nozzle orifice diameter.
16. The flow cytometer according to claim 10 , wherein the sensor is a charge-coupled device (CCD) camera.
17. The flow cytometer according to claim 10 , wherein the width of the flow stream is determined from the images based on droplet volume.
18. The flow cytometer according to claim 10 , wherein the sensor is positioned at the break-off point of the flow stream.