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Published on: August 5, 2009
Delivery of Fluorescent Probes Using Streptolysin O for Fluorescence Microscopy of Living Cells
Kai Wen Teng1,2,3, Pin Ren1,2, Paul R Selvin1,2,3
1Center for Biophysics and Quantitative Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois.
Abstract:
Methods to efficiently deliver fluorophores across the cell membrane are crucial for imaging the dynamics of intracellular proteins using fluorescence. Here we describe a simple protocol for permeabilizing living cells using streptolysin O, a bacterial toxin, which allows transient uptake of fluorescent probes for labeling specific intracellular proteins. The technique is applicable for delivering different classes of fluorescent probes with a molecular weight of <150 kDa, and it is also applicable to a variety of different cell lines. The technique enables the utilization of a broad range of fluorophores for live cell imaging of intracellular proteins. Extended observation of intracellular fluorescence bound to specific proteins is now possible through super-resolution microscopy by using fluorophores that are photostable in "cell-friendly" deoxygenating and reducing conditions. © 2018 by John Wiley & Sons, Inc.
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