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Abstract

Biology

Reconstruction of Single-Cell Innate Fluorescence Signatures by Confocal Microscopy

Published: May 27th, 2020

DOI:

10.3791/61120

1Graduate School of Life and Environmental Sciences, University of Tsukuba, 2Faculty of Life and Environmental Sciences, University of Tsukuba, 3Microbiology Research Center for Sustainability, University of Tsukuba

* These authors contributed equally

Abstract

Described here is confocal reflection microscopy-assisted single-cell innate fluorescence analysis (CRIF), a minimally invasive method for reconstructing the innate cellular fluorescence signature from each individual live cell in a population distributed in a three-dimensional (3D) space. The innate fluorescence signature of a cell is a collection of fluorescence signals emitted by various biomolecules within the cell. Previous studies established that innate fluorescence signatures reflect various cellular properties and differences in physiological status and are a rich source of information for cell characterization and identification. Innate fluorescence signatures have been traditionally analyzed at the population level, necessitating a clonal culture, but not at the single-cell level. CRIF is particularly suitable for studies that require 3D resolution and/or selective extraction of fluorescence signals from individual cells. Because the fluorescence signature is an innate property of a cell, CRIF is also suitable for tag-free prediction of the type and/or physiological status of intact and single cells. This method may be a powerful tool for streamlined cell analysis, where the phenotype of each single cell in a heterogenous population can be directly assessed by its autofluorescence signature under a microscope without cell tagging.

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Keywords Single cell Analysis

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