Common Phenotypes and Solutions - Cell
No Cell Events -- Only marker detected
Most Probable Causes
- No cells in sample -- Prepare a new chip: use concentration as given in the Reagent Kit guide; check cell concentration with a counting chamber; adjust concentration if necessary; visually inspect sample well under microscope to confirm that cells were loaded correctly
- Bead sample not prepared according to the instructions -- Prepare a new chip: refer to the Cell Fluorescence Checkout Kit Guide for proper bead preparation
- Low staining efficiency -- Check staining procedure; always prepare positive and negative controls; your staining protocol may need optimization for you specific cell preparation
Probable Causes
- Cells not resuspended in CB -- Always resuspend cells in CB at an appropriate cell concentration before analysis
- Used dye not compatible with bioanalyzer optics -- For application protocols and recommended staining reagents, refer to available application notes
- Not enough buffer or focusing dye in chip wells -- Prepare a new chip: refer to the Cell Reagent Kit Guide for proper chip preparation; do not leave any wells empty
- Chip not properly primed; unremovable air bubble in chip -- Prepare a new chip: prime the chip according to "Essential Measurement Practices"
- Chip channel clogged -- Prepare a new chip: use cell strainer for clumpy cell sample
Least Probable Causes
- Pipetting error during cell preparation -- Use appropriate pipettes and pipette tips; check calibration of pipette
- Autofocus failure -- Check autofocus using the Diagnostic test, Optical Drive
- Chip contaminated -- Prepare a new chip; wear powder-free gloves only; do not touch the underside of the chip; do not touch the wells of the chip; load the chip immediately after taking it out of its sealed bag
Low Cell Events -- Event number below 400
Most Probable Causes
- Low cell concentration -- Prepare a new chip: use cell concentration of 2 million cells/ml as recommended in the Reagent Kit guide; check cell concentration with a counting chamber; adjust concentration if necessary
- Bead sample not prepared according to the instructions -- Prepare a new chip: refer to the Cell Fluorescence Checkout Kit Guide for proper bead preparation
- Low staining efficiency -- Check staining procedure; always prepare positive and negative staining controls; your staining protocol may need optimization for your specific cell preparation
Probable Causes
- Not enough buffer in buffer well -- Prepare a new chip: refer to the Cell Reagent Kit Guide for proper chip preparation
- No focusing dye in FD-well -- Prepare a new chip: refer to the Cell Reagent Kit Guide for proper chip preparation
- Not enough sample in sample well -- Prepare a new chip: refer to the Cell Reagent Kit Guide for proper chip preparation
- Chip not properly primed; unremovable air bubble in chip -- Prepare a new chip; prime the chip according to the "Essential Measurment Practices"
- Chip channel clogged -- Prepare a new chip: use cell strainer for clumpy cell sample
Least Probable Causes
- Pipetting error during cell preparation -- Use appropriate pipettes and pipette tips; check calibration of pipette
- Autofocus failure -- Check autofocus using the Optical Drive test
- Chip contaminated -- Prepare a new chip; wear powder-free gloves only; do not touch the underside of the chip; do not touch the wells of the chip; load the chip immediately after taking it out of its sealed bag
Poor Fluorescence Intensity
Most Probable Causes
- Low staining efficiency -- Check staining procedure; always prepare positive and negative staining controls; the staining protocol may need optimization for you specific cell preparation
Least Probable Causes
- Autofocus failure -- Check autofocus using the Optical Drive test
- Poor focusing because of decomposed focusing dye -- Prepare a new chip: use fresh focusing dye; protect the focusing dye solution from light
High Events
Most Probable Causes
- Too high cell concentration -- Results may be inaccurate; consider preparing a new chip: use concentration as given in the Reagent Kit guide; check cell concentration with a counting chamber; adjust concentration if necessary
- Bead sample not prepared according to the instructions -- Prepare a new chip: refer to the Cell Fluorescence Checkout Kit Guide for proper bead preparation
Probable Causes
- Wrong assay selected: a conventional assay was selected while the staining was performed on-chip -- Import markers and settings from the corresponding on-chip assay
- Not enough buffer in buffer well -- Prepare a new chip: refer to the Cell Reagent Kit Guide for proper chip preparation
- Not enough sample in sample well -- Prepare a new chip: refer to the Cell Reagent Kit Guide for proper chip preparation
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