Incomplete Red Blood Cell Lysis in Whole Blood

Symptom

Red blood cell debris persists in whole blood samples after lysis protocol, causing high background and interfering with target cell population analysis.

1. 1 Additional washes insufficient to eliminate red blood cell debris using Flow Alternate Protocol
2. 2 0.1% Triton X-100 solution not fresh or improperly prepared, reducing lysis efficiency
3. 3 Deviation from optimized CST Flow protocol steps or reagent concentrations

1. 1 Add up to 3 additional washes between each step to thoroughly remove red blood cell debris
2. 2 Use fresh reagents and prepare new 0.1% Triton X-100 solution immediately before use
3. 3 Closely follow CST Flow Alternate Protocol derived from Chow et al. (2005) Cytometry Part A 67A:4–17, which has been extensively optimized for whole blood

Related Video (3)

Bilibili (China-Accessible Mirrors) ★ 80

Flow Cytometry Experimental Operation in 7 Minutes

"Hands-on flow cytometry protocol including sample preparation, the stage where whole-blood RBC lysis is performed and where this failure occurs."

Bilibili (China-Accessible Mirrors) ★ 75

Flow Cytometry Complete Workflow: Sample to Analysis

"Complete sample-to-analysis workflow with troubleshooting, useful for understanding and correcting sample prep issues like incomplete RBC lysis."

Bilibili (China-Accessible Mirrors) ★ 70

Zhejiang University Senior's Flow Cytometry Hands-On Tutorial

"Detailed step-by-step experimental workflow with direct protocol guidance, helpful for seeing correct sample preparation technique."