Consistent Lysosomal Staining

November 15, 2016

University of Michigan
Pharmaceutical Sciences
Research Fellow

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Company:

Thermo Fisher Scientific

Product Name:

LysoTracker® Blue DND-22

Catalog Number:

L7525

Lysosomes are important cellular compartments that regulate the traffic within the cell of various biochemical entities. Many experiments need a reliable marker for lysosomal staining. While most papers mention the use of Lysotracker Red or Green, we needed a blue fluorescent lysosome stainer to avoid cross-talk between our other fluorescently tagged reagents (whose fluorescence could not be modulated). Upon use of this product as noted from the manufacturer recommended protocol, we got reliable and high staining of lysosomes in our cells (RAW264.7)

Experimental Design and Results Summary

Application

Lysosome staining in live cells

Starting Material

RAW264.7 cells cultured on microscope well-plates or chambers

Protocol Overview

As recommended by manufacturer

Tips

The typical usage recommended is at <= 100 nm. However, we used 1 uM as lower concentrations resulted in extremely light and diffuse staining

Results Summary

As noted in the figure, the staining was very visible and clear. Two issues noted in our use of this product.1. Concentration required is higher than what is noted by the manufacturer.2. The fluorescence bleaching is very evident and fast. Since we only needed the location of the stain to be clear, this was not an issue for us. However, any study that is using this product for quantitative analysis of staining intensity should be extremely careful with their analysis and/or image acquisition method.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Reliable marker of lysosomes. Inexpensive in terms of the number of assays that can be performed with the number of vials in the packaging.

The Bad

Rapid Photobleaching

The Bottom Line

Great for quick visualization and spatial distribution of lysosomes.

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