Related Experiment Video
Updated: May 9, 2026
06:22
Multicolor Flow Cytometry-based Quantification of Mitochondria and Lysosomes in T Cells
Published on: January 9, 2019
Fluorescence flow cytometry methodology to exclude platelet aggregate interference when measuring feline CD4 and CD8
1Research and Development Department, IDEXX Laboratories, Westbrook, ME, USA.
Veterinary Journal (London, England : 1997)
|July 13, 2013
Summary
Platelet aggregate interference in feline lymphocyte analysis can be overcome using CD61 antibody tagging. This improved flow cytometry method accurately quantifies lymphocyte subtypes for disease monitoring in cats.
Area of Science:
- Veterinary Immunology
- Flow Cytometry
- Feline Medicine
Background:
- Clinical monitoring of feline diseases relies on changes in lymphocyte subsets.
- Platelet aggregates commonly interfere with accurate lymphocyte subtype quantification via flow cytometry in whole blood.
Purpose of the Study:
- To develop and validate a flow cytometry method to accurately measure feline lymphocyte subsets by minimizing platelet aggregate interference.
- To improve the clinical utility of flow cytometry for feline disease monitoring.
Main Methods:
- Red blood cells were lysed to isolate lymphocytes.
- Platelets were tagged with anti-CD61AF647 antibody to identify and exclude platelet aggregates.
- Lymphocyte subtypes (CD4%, CD8%, CD8low%, CD4:CD8) were measured using simultaneous 3-color fluorescence flow cytometry.
- Method stability and precision were assessed over 3 days.
Main Results:
- Exclusion of platelet aggregates using CD61AF647 significantly increased CD4%, CD8%, CD8low%, and CD4:CD8 counts (P<0.001).
- The method demonstrated robust stability and precision over 3 days, with low day-to-day coefficients of variation (CVs) for WBC, lymphocyte, CD4, CD8 counts, and CD4:CD8 ratio.
Conclusions:
- CD61 antibody tagging effectively reduces platelet aggregate interference in feline lymphocyte subset analysis.
- This validated flow cytometry method provides a stable and precise approach for quantifying feline lymphocyte subtypes, enhancing their clinical application in disease monitoring.
