Related Experiment Video
Updated: Mar 11, 2026

Visualization of Endoplasmic Reticulum Subdomains in Cultured Cells
Published on: February 18, 2014
A Critical and Comparative Review of Fluorescent Tools for Live-Cell Imaging
Elizabeth A Specht1,2, Esther Braselmann1,2, Amy E Palmer1,2
1Department of Chemistry and Biochemistry, University of Colorado, Boulder, Colorado 80303;
Abstract:
Fluorescent tools have revolutionized our ability to probe biological dynamics, particularly at the cellular level. Fluorescent sensors have been developed on several platforms, utilizing either small-molecule dyes or fluorescent proteins, to monitor proteins, RNA, DNA, small molecules, and even cellular properties, such as pH and membrane potential. We briefly summarize the impressive history of tool development for these various applications and then discuss the most recent noteworthy developments in more detail. Particular emphasis is placed on tools suitable for single-cell analysis and especially live-cell imaging applications. Finally, we discuss prominent areas of need in future fluorescent tool development-specifically, advancing our capability to analyze and integrate the plethora of high-content data generated by fluorescence imaging.
Related Concept Videos
Two-Dimensional Microscopy in Microbiology
Three-Dimensional Microscopy in Microbiology
Super-resolution Fluorescence Microscopy
Confocal Fluorescence Microscopy
Protein Dynamics in Living Cells
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

