Plasma Membrane staining to quantify intracellular biochemistry via fluorescence

June 08, 2015

University of Michigan
Pharmaceutical Sciences
Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Life Technologies

Product Name:

FM® 1-43 Dye (N-(3-Triethylammoniumpropyl)-4-(4-(Dibutylamino) Styryl) Pyridinium Dibromide)

Catalog Number:

T35356

Plasma membrane staining is another way to counterstain for cellular assays apart from nuclear stains. FM 1-43 dye is a highly specific membrane marker that uniformly distributes for a specific fluorescence signal demarcating the intracellular region from extracellular vasculature. However, optimization to obtain repeatedly satisfying quantifiable results can be tricky.

Experimental Design and Results Summary

Application

Fluorescence Microscopy

Starting Material

In vitro cell culture of RAW264.7 cells or MDCK cells

Protocol Overview

Written manual from the company protocol is available. While the overall protocol followed was similar, we used varying concentrations of the dye to optimize the fluorescence signal. The time of incubation can be tricky with a bit of luck required since prolonged incubation would lead to rather saturated staining whereas removing the staining solution from incubation much quicker than required can lead to no staining whatsoever.

Tips

Optimization of protocol is critical for optimized used of this kit

Results Summary

Most of our cell staining experiments worked pretty well with this dye. A representative confocal image is attached clearly showing the plasma membrane staining.

Additional Notes

None

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Summary

The Good

Excellent specificity for the plasma membrane

The Bad

Optimization is key.

The Bottom Line

FM 1-43 is a very specific plasma membrane marker useful for many live cell, in vitro and histological examination via fluorescence microscopic techniques.

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