BD FACSCelesta Software Operation Training

critical

Instrument Optics/Electronics/Fluidics Quality Control Failure

Data quality is inconsistent between runs or gradually deteriorates over time. Instrument performance metrics fall outside acceptable ranges, affecting sensitivity and accuracy of all measurements.

💡 5 · ✓ 6

severe

Incorrect Fluorescence Compensation and Spectral Overlap

Flow cytometry multicolor panels show false-positive signals in channels not expected to be positive. Positive populations appear in multiple fluorescence channels due to spectral overlap between fluorochromes, making accurate population identification impossible.

💡 3 · ✓ 3

severe

Compensation beads saturate detectors causing overflow

Compensation bead populations appear off-scale or saturate detectors, producing signals outside the linear detection range and preventing accurate compensation matrix calculation.

💡 4 · ✓ 4

severe

Rare Cell Population Incorrectly Gated or Missed

Target rare immune cell subsets (e.g., dendritic cells, innate lymphoid cells, hematopoietic progenitors) appear overestimated or masked by abundant terminally differentiated cells. Gating on single markers yields incorrect population percentages (e.g., 4.5% vs. true 1.2% DCs).

💡 4 · ✓ 4

severe

Fluorescence Spillover into Secondary Detectors

One fluorochrome's emission spectra spills over into another detector channel, creating false positive signals. Data appears contaminated with signals that do not represent true marker expression.

💡 4 · ✓ 4

moderate

Suboptimal Scatter Properties and Poor Resolution

Cells display low Forward Scatter (FSC) and Side Scatter (SSC) values. Populations appear poorly defined or compressed in scatter plots.

💡 4 · ✓ 4

moderate

Cell Cycle Phases Not Resolved in DNA Histogram

Histogram for DNA content does not show distinct G0/G1, S, and G2/M phase peaks. Peaks are broad with high coefficients of variation (CVs).

💡 3 · ✓ 3

minor

Excessively High Event Rate During Acquisition

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.

💡 2 · ✓ 2