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Updated: Aug 8, 2026
Multiplexed Fluorescent Immunohistochemical Staining, Imaging, and Analysis in Histological Samples of Lymphoma
Published on: January 9, 2019
Rapid immunotyping of B-cell non-Hodgkin's lymphomas by flow cytometry. A comparison with the standard frozen-section
T E Witzig1, P M Banks, M J Stenson
1Department of Laboratory Medicine, Mayo Clinic, Rochester, Minnesota 55905.
Insights
Flow cytometry (FC) and frozen section (FS) immunotyping (IT) are effective for lymphoma phenotyping. Combining FC and FS IT offers complementary advantages for accurate lymphoma diagnosis.
Area of Science:
- Hematology
- Oncology
- Immunology
Background:
- Accurate immunotyping is crucial for diagnosing and classifying lymphomas.
- Frozen section (FS) and flow cytometry (FC) are established methods for immunophenotyping lymphoid tissues.
- Comparing the strengths and limitations of FS and FC immunotyping (IT) can optimize diagnostic workflows.
Purpose of the Study:
- To compare the advantages of standard frozen section (FS) and flow cytometry (FC) methods for immunotyping (IT) in lymphoma diagnosis.
- To evaluate the concordance and identify causes of discrepancy between FS and FC IT results.
- To assess the utility of FC in detecting light chain restriction in B-cell non-Hodgkin's lymphomas (NHLs).
Main Methods:
- Immunotyping was performed on 218 lymphoma-suspicious biopsies using both FS and FC methods.
- Independent interpretations of FS and FC IT results were compared for concordance.
- Discordant cases were reviewed to identify causes of discrepancy.
- Statistical analysis of kappa and lambda histograms and multiparameter analysis were used to assess light chain restriction in FC.
Main Results:
- FS and FC IT results showed 93% concordance (202/218 cases).
- Seven causes for the 16 discordant cases were identified, including methodologic issues, focal involvement, and FC's enhanced sensitivity.
- FC effectively identified light chain restriction in B-cell NHLs, even when kappa:lambda ratios were inconclusive.
Conclusions:
- Both FS and FC methods are effective for lymphoma phenotyping.
- FS and FC IT possess distinct features that make them complementary for comprehensive lymphoma diagnosis.
- FC offers enhanced sensitivity for detecting light chain restriction, aiding in B-cell NHL classification.
Abstract:
The authors compared immunotyping (IT) results obtained by both standard frozen section (FS) and flow cytometry (FC) methods on 218 biopsies suggestive of lymphoma to learn the advantages of each method. The independent interpretations of the FS and FC IT results were concordant in 93% (202 of 218) of cases. The 16 cases with discordance were reviewed and seven causes for discrepancy found: methodologic problems, focal lymphomatous involvement, more sensitive light chain detection by FC, inadequate sample for FC, interpretation error, sample mislabeling for FC, and unexplained. Eleven of the concordant B-cell non-Hodgkin's lymphomas (NHLs) studied by FC did not have a kappa:lambda ratio of 3 or greater or 0.5 or less and were shown to express light chain restriction by a D-value of 15 or greater with the use of statistical analysis of the kappa and lambda histograms or by multiparameter analysis of large versus small cells. The authors found both methods to be effective for phenotyping lymphomas, however, each has distinct features, making them complementary in their applications.
