Structure of mouse spleen investigated by 7-color fluorescence imaging

Hiromichi Tsurui; Shinichirou Niwa; Sachiko Hirose; Ko Okumura; Toshikazu Shirai

doi:10.1117/12.432481

2 July 2001 Structure of mouse spleen investigated by 7-color fluorescence imaging

Hiromichi Tsurui, Shinichirou Niwa, Sachiko Hirose, Ko Okumura, Toshikazu Shirai

Author Affiliations +

Proceedings Volume 4259, Biomarkers and Biological Spectral Imaging; (2001) https://doi.org/10.1117/12.432481
Event: BiOS 2001 The International Symposium on Biomedical Optics, 2001, San Jose, CA, United States

Abstract

Multi-color fluorescence imaging of tissue samples has been an urgent requirement in current biology. As far as fluorescence signals should be isolated by optical bandpass filter-sets, rareness of the combination of chromophores with little spectral overlap has hampered to satisfy this demand. Additivity of signals in a fluorescence image accepts applying linear unmixing of superposed spectra based on singular value decomposition, hence complete separation of the fluorescence signals fairly overlapping each other. We have developed 7-color fluorescence imaging based on this principle and applied the method to the investigation of mouse spleen. Not only rough structural features in a spleen such as red pulp, marginal zone, and white pulp, but also fine structures of them, periarteriolar lymphocyte sheath (PALS), follicle, and germinal center were clearly pictured simultaneously. The distributions of subsets of dendritic cells (DC) and macrophages (M(phi) ) markers such as BM8, F4/80, MOMA2 and Mac3 around the marginal zone were imagined simultaneously. Their inhomogeneous expressions were clearly demonstrated. These results show the usefulness of the method in the study of the structure that consists of many kinds of cells and in the identification of cells characterized by multiple markers.

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Hiromichi Tsurui, Shinichirou Niwa, Sachiko Hirose, Ko Okumura, and Toshikazu Shirai "Structure of mouse spleen investigated by 7-color fluorescence imaging", Proc. SPIE 4259, Biomarkers and Biological Spectral Imaging, (2 July 2001); https://doi.org/10.1117/12.432481

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