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Updated: Aug 8, 2026
Techniques for the Analysis of Extracellular Vesicles Using Flow Cytometry
Published on: March 17, 2015
Techniques to improve flow cytometric detection of light chain restriction
G R Shaw1, E Gonering, S Koller
1Marshfield Clinic, WI 54449, USA.
Objective:
To compare 3-color flow cytometry (using a permeabilization step to detect cytoplasmic immunoglobulin in selected cases) with 2-color flow cytometry in the detection of light chain restriction (LCR).
Design:
Analysis of clinical specimens submitted for lymphocyte immunophenotyping using both methods.
Setting:
Marshfield Laboratories serving Saint Joseph's Hospital (525 beds) and the Marshfield Health Care Network.
Main Outcome Measure(S):
Sensitivity and specificity for detecting LCR in B-cell neoplasms. Final diagnosis based on review of clinical, laboratory and histologic data.
Results:
Of 61 specimens, the 3-color method yielded better sensitivity, detecting LCR in 30 of 39 cases of B-cell neoplasms (77%) versus 16 of 39 (41%) for the 2-color method (P < 0.001). Both methods had comparable specificity (95-100%). The 3-color cytoplasmic technique identified another 4 cases yielding an overall sensitivity of 87% for a 2-tiered testing strategy.
Conclusion:
A 3-color surface technique, backed up by a permeabilization step in selected cases, provides a cost-effective and sensitive technique for detecting LCR.
