Oct 25, 2024
ZSGreen x TH Colocalization Analysis
- 1University of California San Diego;
- 2University of California, San Diego
Protocol Citation: Grace Kollman, Thomas Hnasko 2024. ZSGreen x TH Colocalization Analysis. protocols.io https://dx.doi.org/10.17504/protocols.io.3byl4we3zvo5/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: October 24, 2024
Last Modified: October 25, 2024
Protocol Integer ID: 110865
Abstract
Colocalization analysis of TH and ZSGreen using immunohistochemistry
Identify and draw subregion borders on the image in Zen.
Going subregion at a time, use the event tool in Zen software to mark cells in which ZSGreen signal appears to be present in at least 50% of the cell as ZSGreen positive. ZSGreen signal should also be associated with a single nuclei shown with DAPI.
If a cell is on the border of a region the cell should be counted toward the region which
contains at least 50% of the cell.
After identifying ZSGreen positive cells, use the event tool to mark any cells expressing TH and ZSGreen in a similar shape as double positive.
Counts are organized and percentages are calculated using Microsoft excel.