Methods and apparatus for real-time detection and clearing of a clog
View Patent ↗A flow cytometer apparatus and methods for detecting and clearing a clog therein are disclosed. An example method for detecting a clog may include (i) detecting, via a fault detection system of a flow cytometer, a first plurality of events associated with a first aliquot from a first sample well, (ii) determining a count of the first plurality of events associated with the first aliquot, (iii) determining whether the count of the first plurality of events is below a minimum count tolerance and (iv) (a) if the count of the first plurality of events is below the minimum count tolerance, then determining that the flow cytometer has a clog, (b) if the count of the first plurality of events is equal to or above the minimum count tolerance, then detecting a second plurality of events associated with a second aliquot from a second sample well.
1. A method, comprising:
providing a flow cytometer system comprising (a) a flow cell having a first end and a second end, (b) a sample fluidic pathway having a first end and a second end, wherein the second end of the sample fluidic pathway is coupled to the first end of the flow cell, (c) a sample probe coupled to the first end of the sample fluidic pathway, (d) a sample pump in fluid communication with the sample fluidic pathway, (e) a waste line having a first end and a second end, wherein the first end of the waste line is coupled to the flow cell and (f) a waste pump in fluid communication with the waste line;
activating the waste pump to apply negative pressure to one or more of the waste line, the flow cell, the sheath fluidic pathway and the sample fluidic pathway;
activating the sample pump;
cycling the sample probe into and out of a decontamination solution reservoir and driving a decontamination fluid, via the sample pump, through one or more of the flow cell, the sample fluidic pathway and the waste line, thereby clearing a clog, wherein cycling the sample probe into and out of the decontamination solution reservoir occurs for a period of time ranging from about two minutes to about five minutes.
2. The method of claim 1 , further comprising:
cycling the sample probe into and out of a cleaning solution reservoir; and
driving a cleaning fluid, via the sample pump, through one or more of the flow cell, the sample fluidic pathway or the waste line, thereby cleaning away the decontamination fluid.
3. The method of claim 2 , further comprising:
cycling the sample probe into and out of a cell-compatible fluid reservoir; and
driving cell-compatible fluid through one or more of the waste line, the flow cell and the sample fluidic pathway, thereby removing any remaining cleaning fluid.
4. The method of claim 3 , further comprising:
during the cycling of the sample probe into and out of the cell-compatible fluid reservoir, pumping a plurality of cell-compatible fluid samples separated by air samples through at least the sample fluidic pathway.
5. The method of claim 2 , further comprising:
during the cycling the sample probe into and out of the cleaning solution reservoir, pumping a plurality of cleaning fluid samples separated by air samples through at least the sample fluidic pathway.
6. The method of claim 3 , wherein cycling the sample probe into and out of the cell-compatible fluid reservoir occurs for a period of time ranging from about two minutes to about five minutes.
7. The method of claim 2 , wherein cycling the sample probe into and out of the cleaning solution reservoir occurs for a period of time ranging from about two minutes to about five minutes.
8. The method of claim 1 , further comprising:
providing a sheath fluidic pathway coupled to the first end of the flow cell and a sheath pump in fluid communication with the sheath fluidic pathway;
activating the sheath pump; and
driving a sheath fluid through one or more of the sheath fluidic pathway, the flow cell and the waste line.
9. The method of claim 1 , further comprising:
during the cycling of the sample probe into and out of the decontamination solution reservoir, pumping a plurality of decontamination fluid samples separated by air samples through the sample fluidic pathway.
10. The method of claim 1 , further comprising:
prior to activating the waste pump, ceasing a sampling operation and a data acquisition operation.
11. The method of claim 1 , further comprising:
providing a three-port valve coupled to the waste line between the flow cell and the waste pump, wherein the three-port valve has a first port, a second port and a third port, wherein the first port of the three-port valve is arranged in series with the second port of the three-port valve such that the first-port is arranged closer to the flow cell than the second port, and wherein the third port of the three-port valve is configured to communicate with atmosphere; and
wherein cycling the sample probe into and out of the decontamination solution reservoir comprises:
(a) operating the flow cytometer system in a forward mode;
(b) deactivating the waste pump and the sample pump;
(c) holding a plurality of decontamination fluid samples separated by air samples in at least the sample fluidic pathway;
(d) closing the second port of the three-port valve and opening the third port of the three-port valve;
(e) operating the sample pump in a reverse-mode and flowing the plurality of decontamination fluid samples in reverse through at least the sample fluidic pathway.
12. The method of claim 11 , further comprising cycling the sample probe into and out of a cleaning solution reservoir, wherein cycling the sample probe into and out of the cleaning solution reservoir comprises:
(a) operating the flow cytometer system in the forward mode;
(b) deactivating the waste pump and the sample pump;
(c) holding a plurality of cleaning fluid samples separated by air samples in the sample fluidic pathway;
(d) closing the second port of the three-port valve and opening the third port of the three-port valve; and
(e) operating the sample pump in the reverse-mode and flowing the plurality of decontamination fluid samples in reverse through at least the sample fluidic pathway.
13. The method of claim 12 , further comprising cycling the sample probe into and out of a cell-compatible fluid reservoir, wherein cycling the sample probe into and out of the cell-compatible fluid reservoir comprises:
(a) activating the sheath pump; and
(b) operating the flow cytometer system in the forward mode.
14. The method of claim 1 , wherein the sample probe is positioned over a back flush waste fluid reservoir when the sample pump is operating in a reverse-mode.