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Published on: November 28, 2015
Enhanced detection of malignant lymphoma in cerebrospinal fluid by multiparameter flow cytometry
W G Finn1, L C Peterson, C James
1Department of Pathology, Northwestern University Medical School, Chicago, Illinois, USA.
Insights
Flow cytometry enhances detection of malignant lymphoma in cerebrospinal fluid (CSF) samples. This sensitive method improves diagnostic accuracy by identifying small lymphoid clones, especially in low-cell-count CSF specimens.
Area of Science:
- Hematology
- Clinical Pathology
- Immunology
Background:
- Cerebrospinal fluid (CSF) immunophenotyping by flow cytometry is not widely established.
- Standard microscopic examination has limitations in detecting malignant lymphoma in CSF.
Purpose of the Study:
- To optimize flow cytometric detection of malignant lymphoma in CSF samples.
- To evaluate the utility of 3- and 4-color flow cytometry for CSF analysis.
- To compare flow cytometry results with standard microscopic examination.
Main Methods:
- Immunophenotyping of 36 consecutive CSF samples using 3- and 4-color flow cytometry.
- Selection of lymphoid cells based on fluorescence and light scatter properties.
- Comparison of flow cytometry findings with results from standard microscopic examination.
Main Results:
- Adequate lymphoid cell analysis was achieved in 27 out of 36 CSF samples.
- Flow cytometry detected lymphoma in 10 cases, with 3 cases identified exclusively by this method.
- The combination of flow cytometry and morphology increased lymphoma detection by 43% compared to morphology alone.
- Flow cytometry identified lymphoid clones comprising less than 1% of total cells.
Conclusions:
- Multicolor flow cytometry is a rapid and sensitive technique for detecting lymphoma in paucicellular CSF samples.
- Flow cytometry significantly enhances the diagnostic yield for malignant lymphoma in CSF.
- Further studies are needed to determine the clinical significance of detecting small lymphoid clones in CSF.
Abstract:
Immunophenotyping by flow cytometry has not been widely applied to cerebrospinal fluid (CSF) analysis. We attempted to optimize flow cytometric detection of malignant lymphoma in CSF samples by the routine use of 3- and 4-color flow cytometry, with specific selection of lymphoid cells by fluorescence vs 90 degrees light scatter gating. Thirty-six consecutive CSF samples were immunophenotyped by flow cytometry, and the results were compared with those of standard microscopic examination. Lymphoid events were adequate for analysis in 27 of the 36 samples. Each of the 9 unsuccessful samples was more than 24 hours old at analysis or contained fewer than 1 x 10(4) total cells (< or =1 cell/microL). Lymphoma was detected in 10 of the remaining 27 cases. Six lymphomas were detected by morphology and flow cytometry, 1 only by morphologic examination, and 3 only by flow cytometry. Therefore, the combination of flow cytometry and morphologic examination enhanced the detection by 43% over morphologic examination alone. Flow cytometry permitted the detection of lymphoid clones totaling less than 1% of total cells. Multicolor flow cytometry is a rapid and sensitive technique that enhances detection of lymphoma in paucicellular CSF samples. Given the great sensitivity of flow cytometry, future studies will be necessary to assess the significance of detecting small lymphoid clones in this setting.
