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Updated: Jul 17, 2026
A Fluorescence-based Lymphocyte Assay Suitable for High-throughput Screening of Small Molecules
Published on: March 10, 2017
Identification and quantification of high fluorescence-stained lymphocytes as antibody synthesizing/secreting cells
J Linssen1, V Jennissen, J Hildmann
1Sysmex Europe GmbH, 22848 Norderstedt, Germany. Linssen.jo@sysmex-europe.com
Insights
The SYSMEX XE-2100 high fluorescence lymphocytes count accurately identifies antibody-synthesizing cells (ASC) in reactive diseases. This automated method offers a reliable tool for screening and monitoring patients with suspected infections.
Area of Science:
- Hematology
- Immunology
- Clinical Chemistry
Background:
- The SYSMEX XE-2100 offers a high fluorescence lymphocytes (HFL) count in its leucocyte differential channel.
- Identifying HFL cells as antibody-synthesizing or -secreting cells (ASC) in reactive diseases is crucial for diagnosis.
Purpose of the Study:
- To classify and quantify HFL-count as ASC in reactive diseases.
- To evaluate HFL-count's accuracy against other cell populations and diagnostic methods.
Main Methods:
- Analysis of 85 patients using the SYSMEX XE-2100.
- Comparison with automated image analysis (Cellavision Diffmaster 96) and immunophenotyping (BD FACSCalibur).
Main Results:
- HFL-count demonstrated high reproducibility and linearity (R(2) = 0.99).
- Excellent correlation found between HFL-count and ASC identified by peripheral blood smear (R(2) = 0.900) and immunophenotyping (R(2) = 0.897).
- No significant correlation was observed with monocytes or immature granulocytes.
Conclusions:
- The SYSMEX XE-2100 HFL-count precisely identifies and quantifies ASC (activated B-lymphocytes).
- This automated count serves as a potential screening and monitoring tool for suspected infections.
- Further research is needed to explore the full clinical utility of HFL (ASC) counts in diagnosing inflammation, infection, or sepsis.
Objectives:
The aim of this study was to classify and quantify the high fluorescence lymphocytes area (HFL-count) from the SYSMEX XE-2100 leucocyte differential channel as antibody-synthesizing or -secreting cells (ASC, plasma cells or lymphoplasmacytoid cells) in reactive diseases. To unequivocally identify the HFL cells, all possibly eligible cell populations have been investigated: activated B-lymphocytes, activated T-lymphocytes, large granular lymphocytes (LGL), activated monocytes, and immature granulocytes.
Methods:
In total, 85 patients were analyzed on the XE-2100 and compared with the automated image analysis system Cellavision Diffmaster 96 based on artificial neural network and immunophenotyping method with the BD FACSCalibur.
Results:
Reproducibility tests for HFL demonstrated a mean coefficient of variation of 13.9% for very low results and 1.5% for high results. The linearity data showed a good correlation (R(2) = 0.99) between expected and measured HFL. The comparison with possibly eligible cell populations showed no significant correlation between activated monocytes and immature granulocytes, with most immature granulocytes (promyelocyte I or II), natural killer cells or LGLs, activated T-lymphocytes, and sub-T-lymphocytes populations. However, for activated B-lymphocytes an excellent significant correlation with the peripheral blood smear, and the immunophenotyping method has been found with R(2) = 0.900, P < 0.001 and R(2) = 0.897, P < 0.001, respectively. The slope of 1.1 and intercept of minus 5 cells/microL of the regression equation between HFL-count and ASC (smear) do indicate an excellent quantification of the HFL-count, as well.
Conclusion:
The fully automated SYSMEX XE-2100 HFL-count identifies and quantifies the ASC cells (activated B-lymphocytes) with high precision and reliability in patients without hematology system diseases, thus providing a potential screening and monitoring tool for any patient with suspected infection. Additional studies are required to comprehend in more detail the full clinical utility of an HFL (ASC) count as a potential diagnostic indicator of inflammation, infection, or sepsis.
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