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Updated: Aug 11, 2026
Automated Slide Scanning and Segmentation in Fluorescently-labeled Tissues Using a Widefield High-content Analysis System
Published on: May 3, 2018
Automated four-color analysis of leukocytes by scanning fluorescence microscopy using quantum dots
József Bocsi1, Dominik Lenz, Anja Mittag
1Department of Pediatric Cardiology, Heart Center, University Leipzig, Leipzig, Germany. jobocsi@web.de
Background:
Scanning fluorescence microscope (SFM) is a new technique for automated motorized microscopes to measure multiple fluorochrome labeled cells (Bocsi et al., Cytometry A 2004, 61:1-8).
Aims:
We developed a four-color staining protocol (DNA, CD3, CD4, and CD8) for the lymphocyte phenotyping by SFM.
Methods:
Organic (Alexa488, FITC, PE-Alexa610, CyChrom, APC) and inorganic (quantum dot (QD) 605 or 655) fluorochromes were used and compared in different combinations. Measurements were performed in suspension by flow cytometer (FCM) and on slide by SFM.
Results:
Both QDs were detectable by the appropriate Axioplan-2 and FCM filters and the AxioCam BW-camera. CD4/CD8 ratios were highly correlated (P = 0.01) between the SFM and FCM.
Conclusion:
Automated SFM is an applicable tool for CD4/CD8 ratio determination in peripheral blood samples with QDs.
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