Cell counting

daniel.dautan daniel, Per Svenningsson

Published: 2024-07-13 DOI: 10.17504/protocols.io.14egn66rql5d/v1

Abstract

Used for counting cells labeled with immunofluorescent markers in mouse brain sections. Sections should be stained, mounted, and imaged with high resolution (2048 x 2048 scanning).

Steps

1.

Using 2-3 sections to encompass the brain region of interest, acquire high resolution images (2048 x 2048) using tile scanning and z-stack acquisition.

2.

Import images into ImageJ.

3.

Apply maximum average projection and color adjust as needed, making sure to apply the same settings to all images.

4.

For each image, use the multi-section tool in ImageJ to manually quantify the positively-labeled cells of interest. Multiple channels can be viewed to assess cells labeled with multiple markers.

5.

Measure the region's area using the selection tool. Use this value to derive the relative density of cells.

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