A Review of Reagents for Fluorescence Microscopy of Cellular Compartments and Structures, Part III: Reagents for Actin, Tubulin, Cellular Membranes, and Whole Cell and Cytoplasm

Abstract

AbstractNon‐antibody commercial fluorescent reagents for imaging of cytoskeletal structures have been limited primarily to labeling tubulin and actin, with the key factor in choice based mainly on whether cells are live or fixed and permeabilized. A wider range of options exists for cell membrane dyes, and the choice of reagent primarily depends on the preferred localization in the cell (i.e., all membranes or only the plasma membrane) and usage (i.e., whether the protocol involves fixation and permeabilization). For whole‐cell or cytoplasmic imaging, the choice of reagent is determined mostly by the length of time that the cells need to be visualized (hours or days) and by fixation status. Presented here is a discussion on choosing commercially available reagents for labeling these cellular structures, with an emphasis on use for microscopic imaging, with a featured reagent, recommended protocol, troubleshooting guide, and example image for each structure. © 2023 Wiley Periodicals LLC.Basic Protocol 1: Actin labelingBasic Protocol 2: Wheat germ agglutinin conjugates for plasma membrane labelingBasic Protocol 3: Labeling tubulin microtubules with Tubulin Tracker Deep RedBasic Protocol 4: Labeling whole cells or cytoplasm with 5(6)‐CFDA SE