Jul 12, 2024

Public workspaceCell counting

DOI
dx.doi.org/10.17504/protocols.io.14egn66rql5d/v1
  • 1Department of Clinical Neuroscience, Karolinska Institutet, 171 76 Stockholm, Sweden;
  • 2Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, MD 20815, USA
Eileen Ruth Torres
Open access
DOI: dx.doi.org/10.17504/protocols.io.14egn66rql5d/v1
Protocol Citation: daniel.dautan daniel, Per Svenningsson 2024. Cell counting. protocols.io https://dx.doi.org/10.17504/protocols.io.14egn66rql5d/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: June 04, 2024
Last Modified: July 12, 2024
Protocol Integer ID: 101203
Keywords: ASAPCRN, neurons, apoe, chat
Funders Acknowledgement:
Aligning Science Across Parkinson's
Grant ID: 020608
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).
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.
Import images into ImageJ.
Apply maximum average projection and color adjust as needed, making sure to apply the same settings to all images.
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.
Measure the region's area using the selection tool. Use this value to derive the relative density of cells.