Excessively High Event Rate During Acquisition

Symptom

Flow cytometer records excessive events per second (>10,000/sec), leading to coincidence errors where multiple cells pass through the laser simultaneously. Data shows abnormal event clustering and unreliable fluorescence measurements.

1. 1 Cell concentration too high (above 1x10^6 cells/mL), causing multiple cells to be analyzed simultaneously
2. 2 Sample overcorrection during preparation, providing excessive margin for error

1. 1 Dilute sample to optimal concentration between 1x10^5 and 1x10^6 cells/mL
2. 2 Re-count cells using hemocytometer or automated cell counter before preparing final sample; aim for mid-range concentration to balance event rate and acquisition time

Related Video (3)

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"Complete flow cytometry workflow from sample preparation through instrument operation and troubleshooting, directly covering the context where high event rate/concentration issues arise."

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"Hands-on protocol including sample preparation and parameter adjustment, where cell concentration and event rate settings would be addressed."

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"Practical instrument operation tutorial demonstrating parameter settings and acquisition on the flow cytometer, relevant to recognizing/handling event rate problems."