Hawley Teresa S, Hawley Robert G, Telford William G
Flow Cytometry Core Facility, Laboratory of Genome Integrity, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland.
Department of Anatomy and Regenerative Biology, George Washington University School of Medicine and Health Sciences, Washington, D.C.
Curr Protoc Cytom. 2017 Apr 3;80:9.12.1-9.12.20. doi: 10.1002/cpcy.17.
Fluorescent proteins have become standard tools for cell and molecular biologists. The color palette of fluorescent proteins spans the ultraviolet, visible, and near-infrared spectrum. Utility of fluorescent proteins has been greatly facilitated by the availability of compact and affordable solid state lasers capable of providing various excitation wavelengths. In theory, the plethora of fluorescent proteins and lasers make it easy to detect multiple fluorescent proteins simultaneously. However, in practice, heavy spectral overlap due to broad excitation and emission spectra presents a challenge. In conventional flow cytometry, careful selection of excitation wavelengths and detection filters is necessary. Spectral flow cytometry, an emerging methodology that is not confined by the "one color, one detector" paradigm, shows promise in the facile detection of multiple fluorescent proteins. This chapter provides a synopsis of fluorescent protein development, a list of commonly used fluorescent proteins, some practical considerations and strategies for detection, and examples of applications. © 2017 by John Wiley & Sons, Inc.
荧光蛋白已成为细胞和分子生物学家的标准工具。荧光蛋白的调色板涵盖紫外线、可见光和近红外光谱。紧凑且价格合理的固态激光器能够提供各种激发波长，极大地促进了荧光蛋白的应用。理论上，大量的荧光蛋白和激光器使得同时检测多种荧光蛋白变得容易。然而，在实际操作中，由于宽激发光谱和发射光谱导致的严重光谱重叠带来了挑战。在传统流式细胞术中，需要仔细选择激发波长和检测滤光片。光谱流式细胞术是一种不受“一种颜色，一个检测器”模式限制的新兴方法，在轻松检测多种荧光蛋白方面显示出前景。本章提供了荧光蛋白发展的概述、常用荧光蛋白列表、一些检测的实际考虑因素和策略以及应用示例。© 2017约翰威立父子公司版权所有
