CytoPainter LysoOrange Indicator Reagent Before Fixation

Idival Research Institute
Microbiology
Principal Investigator

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Company:

Abcam

Product Name:

CytoPainter LysoOrange

Catalog Number:

ab176827

We are always on the lookout for fluorescent conjugates to stain cellular components in different primary cell lines and immortalized cells. In this case, we used CytoPainter LysoOrange to stain lysosomes in human blood granulocytes and track their biodistribution in live cells. Furthermore, although it is recommended for live cell imaging, CytoPainter retain its fluorescence characteristics after fixation, and the cells can be stained with other dyes or antibodies and observed by confocal laser scanning microscopy.

Experimental Design and Results Summary

Application

Immunofluorescence

Starting Material

Human blood granulocytes

Protocol Overview

The fixation method should be selected on the basis of the experimental requirements. Fixing cultured cells in 3-3.4% paraformaldehyde after live-cell imaging experiments also results in excellent staining with CytoPainter LysoOrange. Storage of CytoPainter LysoOrange: Always protected from light at -20°C (stock solution). We dilute the stock solution to the final working concentration in the growth medium directly.

Tips

None

Results Summary

When we studied lysosomes in human granulocites by fluorescence staining and live-cell imaging, we observed an excellent staining with very good signal and low background at 1:500 dilution, and also after fixation with paraformaldehyde. Figure 1. Image of primary human granulocytes stained in vivo with CytoPainter LysoOrange in a µ-slide using live-cell imaging (A) and after fixation, using a confocal microscope (B-C). Red (LysoOrange), Blue (DAPI). Scale bars A, 20µm, B and C, 5µm

Additional Notes

Detection: Live-cell imaging (545-575 nm excitation, 595-620-nm emission filter). CLSM: 561 nm excitation filter and 570-620 nm emission photomultiplier.

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Summary

The Good

Quality; easy to use. Very good performance on fixed cells.

The Bad

None

The Bottom Line

Very good staining and reproducibility.

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