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Identification of B and T cells in human spleen sections by infrared microspectroscopic imaging
Author(s) -
Krafft Christoph,
Salzer Reiner,
Soff Gerhard,
MeyerHermann Michael
Publication year - 2005
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.20117
Subject(s) - stain , staining , infrared , infrared microscopy , pathology , infrared spectroscopy , immunohistochemistry , chemistry , resolution (logic) , primary and secondary antibodies , microbiology and biotechnology , spleen , antibody , microscopy , biology , medicine , immunology , optics , physics , organic chemistry , artificial intelligence , computer science
Background Infrared spectroscopy probes the chemical composition and molecular structure of complex systems such as tissue and cells. Infrared spectroscopic imaging combines this spectral information with lateral resolution near the single‐cell level. We analyzed whether this method is competitive with classic immunohistochemical methods for immunologic tissue and cells. Methods We recorded infrared microspectroscopic mapping datasets with a 90‐ × 90‐μm 2 aperture from a 3‐ × 3‐mm 2 unstained tissue area of human spleen. A secondary follicle containing a germinal center and a T zone were studied in more detail by infrared microspectroscopic imaging with lateral resolution near 5 μm. The results were compared with consecutive sections stained by immunoglobulin D antibodies. T and B lymphocytes were extracted from human blood and served as independent test samples. Results Cluster analysis of infrared datasets produced images that distinguished anatomical features such as primary and secondary follicles, T zones, arteries, and spleen red pulp. The assignments could be confirmed in consecutive sections by immunohistochemical staining. Main spectral variances between T and B lymphocytes in high‐resolution measurements were attributed to specific spectral contributions of DNA and cytosol. Conclusions Sensitivity and specificity of the infrared based methods are comparable to those of standard staining procedures for identification of B and T cells. However, infrared spectroscopic imaging can offer advantages in velocity, data throughput, and standardization because of minimal sample preparation. The results emphasize the potential of infrared spectroscopy as an innovative tool for the distinction of cell types, in particular in immunologic tissue. © 2005 Wiley‐Liss, Inc.