📋 Workflow
Full workflow, products add-to-cart directly1
Sample preparation
1 hPrepare single-cell suspension, count and adjust density, cells should be monodisperse
💡 Tip Remove clumps and debris
⚠️ Caution Viability affects staining and scatter
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2
Antibody staining
1 hAdd fluorochrome-conjugated antibodies, incubate at 4°C in dark, wash off unbound
💡 Tip Include isotype and single-stain controls
⚠️ Caution Protect from light throughout
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3
Acquisition
1 hSet lasers and fluorescence channels, adjust voltages, acquire and record events
💡 Tip Run controls first to set compensation
⚠️ Caution Moderate flow rate prevents doublets
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4
Data analysis
1 hUse software to gate out debris/doublets, quantify subset percentages and intensities
💡 Tip Consistent gating ensures comparability
⚠️ Caution Document gating for reproducibility
🧪 Materials for this step (click to shop by spec/brand)
⚠️ 5 Common Beginner Mistakes
- ① Clumping causes doublet events skewing analysis
- ② Wrong compensation causes fluorescence spillover artifacts
- ③ EDTA in buffer affects some markers
- ④ Improper antibody amount causes weak signal or non-specific binding
- ⑤ Photobleaching attenuates signal
❓ FAQ
+How to choose fluorochrome panels?
Choose spectrally separated, low-compensation combinations matched to laser setup.
+What is gating?
Selecting cell subsets by scatter/fluorescence, excluding debris and doublets.
+Can cells be sorted simultaneously?
FACS can sort cells by gating into separate populations for downstream culture/analysis.