Gives A Good Contrast For Mouse Paraffin Embedded Sections With Immufluorescence

SUNY Downstate Medical Center
Department of Cell Biology
PhD Candidate (Graduate Student)

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Company:

Thermo Scientific

Product Name:

HOECHST 33342

Catalog Number:

62249

We are studying how learning process is altered in a mouse model with cobalamin deficiency. To further understand it's underlying cause, we studied changes in the expression levels of AMPA receptors using immunofluorescence. We used HOECHT as a counter stain for this technique.

Experimental Design and Results Summary

Application

Immunohistochemistry

Starting Material

Paraffin embedded mouse brain sections

Protocol Overview

After incubation with secondary Ab, sections were washed in Modified Dulbecco's PBS containing 8mM sodium phosphate, 2mM potassium phosphate, 140mM sodium chloride, 10mM potassium chloride, pH 7.4. HOECHST staining was performed according to manufacturer's protocol.

Tips

Make sure to perform the counter-staining in dark

Results Summary

We got nuclei stained for the mouse sections (as shown in the image). It was helpful in locating hippocampus under a fluorescence microscope which was used to determine region specific expression of our protein of interest.

DOI or PMID #

28545069

Additional Notes

N/A

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Summary

The Good

Easy to use and doesn't leak

The Bad

Fades pretty fast

The Bottom Line

It is a good counter-stain and doesn't need any special treatment on the sections to make it work.

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