Methods and systems for collecting cells of a biological specimen
View Patent ↗Methods and systems for collecting cells on a filter disposed in a specimen fluid containing suspended cells of a biological specimen. A short vacuum pulse is applied across a filter to sip specimen fluid. A percentage or portion of the filter surface area covered by cells is determined is representative of or correlates to the density of cells in the specimen fluid. A maximum vacuum amplitude and/or duration are determined utilizing the determined filter coverage. A longer, continuous vacuum or slurp is applied across the filter to collect cells on the filter while limiting the amplitude and/or duration of the slurp based at least in part upon the determined maximum vacuum amplitude and/or duration.
1. A method for collecting cells on a filter, comprising:
positioning the filter in a specimen fluid containing suspended cells of a biological specimen;
determining a maximum rate of aspiration of the specimen fluid across the filter by
applying one or more initial vacuum pulses across the filter to draw specimen fluid across the filter such that the cells cover at least a portion of the filter,
determining a filter coverage quantity as a result of the one or more initial vacuum pulses, and
selecting a maximum rate of aspiration from a pre-determined data set corresponding to the determined filter coverage quantity, wherein the data set comprises a range of maximum rates of aspiration corresponding to pre-selected filter coverage quantities;
selecting from the data set at least one of a maximum amplitude of vacuum and a maximum duration of vacuum corresponding to the determined maximum rate of aspiration; and
applying a further vacuum pulse across the filter to collect cells on the filter while controlling an amplitude and a duration of the further vacuum to correspond to the at least one of the selected maximum amplitude of vacuum and the selected maximum duration of vacuum.
2. The method of claim 1 , wherein determining the maximum rate of aspiration further comprises
performing pulsatile aspiration of the specimen fluid across the filter during the application of the one or more initial vacuum pulses; and
determining the filter coverage quantity during the pulsatile aspiration.
3. The method of claim 2 , further comprising
applying a plurality of initial vacuum pulses to perform intermittent or periodic pulsatile aspiration of specimen fluid across the filter; and
determining the filter coverage quantity during each pulsatile aspiration of the specimen fluid and choosing one of the respective filter coverage quantities.
4. The method of claim 3 , wherein applying a plurality of initial vacuum pulses further comprises applying three initial vacuum pulses to the filter to perform three pulsatile aspirations of the specimen fluid, and wherein the filter coverage quantity is determined during the third pulsatile aspiration.
5. The method of claim 1 , wherein the duration of the further vacuum pulse is substantially longer than a duration of any of the one or more initial vacuum pulses.
6. The method of claim 1 , wherein the duration of each of the initial vacuum pulses is less than about two seconds, and the duration of the further vacuum pulse is greater than about five seconds.
7. The method of claim 1 , wherein the amount of specimen fluid flowing through the filter during application of the further vacuum pulse is substantially greater than the amount of specimen fluid flowing through the filter during application of any of the one or more initial vacuum pulses.
8. The method of claim 1 , wherein the amplitude of the further vacuum pulse increases for a time period of about five to ten seconds, and wherein the amplitude of each of the one or more initial vacuum pulses substantially decays within about two seconds.
9. The method of claim 1 , wherein an area bounded by a curve representing the amplitude of the further vacuum pulse over time is substantially greater than an area bounded by a curve representing an amplitude of any of the one or more initial vacuum pulses over time.
10. The method of claim 1 , further comprising
applying an additional vacuum pulse through the filter after application of the further vacuum pulse, and
collecting additional cells on the filter until a pre-determined filter coverage quantity is reached, the additional vacuum pulse having a duration substantially shorter than the duration of the further vacuum pulse.
11. The method of claim 1 , wherein the filter coverage quantity is determined utilizing a rate of decay of the initial vacuum pulse as cells are collected on the filter.
12. The method of claim 1 , wherein filter coverage quantity is determined utilizing a flow sensor to detect a change of air flow through the filter as cells are collected on the filter.
13. The method of claim 1 , the further vacuum pulse being generated by low duty cycle vacuum pulses having durations that are substantially shorter than a duration of any of the one or more initial vacuum pulses.
14. The method of claim 13 , the low duty cycle vacuum pulses each having a duration on an order of milliseconds, and any of the one or more initial vacuum pulses having a duration on an order of seconds.
15. The method of claim 14 , the low duty cycle vacuum pulses having a respective duty cycle of about 3% to about 15%.
16. A method for collecting cells on a filter, the method comprising:
positioning the filter in a specimen fluid containing suspended cells of a biological specimen;
applying a series of initial vacuum pulses to cause pulsatile aspiration of the specimen fluid across the filter such that the cells cover at least a portion of the filter;
determining a maximum rate of aspiration of the specimen fluid across the filter based at least in part upon
a filter coverage quantity determined as a result of the applied pulsatile aspiration of the specimen fluid across the filter, and
a maximum rate of aspiration selected from a pre-determined data set that corresponds to the determined filter coverage quantity;
determining an amplitude and a duration of a further vacuum pulse to be applied across the filter based at least in part upon at least one of a maximum amplitude of vacuum and a maximum of duration of vacuum selected from the data set that correspond to the determined maximum rate of aspiration; and
applying the further vacuum pulse across the filter to collect cells on the filter while controlling the amplitude and the duration of the further vacuum pulse to correspond to the at least one of the maximum amplitude of vacuum and the maximum duration of vacuum.
17. The method of claim 16 , wherein applying the series of initial vacuum pulses to cause pulsatile aspiration of the specimen fluid across the filter comprises applying a plurality of vacuum pulses to cause intermittent or periodic pulsatile aspiration of specimen fluid across the filter.
18. The method of claim 17 , wherein the filter coverage quantity is determined by determining a respective filter coverage quantity during each pulsatile aspiration of the specimen fluid, and selecting one of the respective determined filter coverage quantities.
19. The method of claim 16 , wherein the filter coverage quantity is determined utilizing a rate of decay of any one of the pulses in the series of initial vacuum pulses as cells collect on the filter.
20. The method of claim 16 , wherein the filter coverage quantity is determined by utilizing a flow sensor to detect a change of air flow through the filter as cells collect on the filter.