Jul 12, 2024

Public workspaceImaging- Confocal

DOI
dx.doi.org/10.17504/protocols.io.j8nlk88y1l5r/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.j8nlk88y1l5r/v1
Protocol Citation: daniel.dautan daniel, Per Svenningsson 2024. Imaging- Confocal. protocols.io https://dx.doi.org/10.17504/protocols.io.j8nlk88y1l5r/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: 101201
Keywords: ASAPCRN, imaging, mouse brain
Funders Acknowledgement:
Aligning Science Across Parkinson's
Grant ID: 020608
Abstract
Protocol for imaging using confocal microscope. Sections for analysis should be mounted on slides, stained for appropriate markers, and coverslipped. This protocol is using a Carl Zeiss LSM 880 confocal microcope.
Using the confocal microscope, capture images at 10x magnification using a resolution of either 1024x1024 or 2048x2048.
If needed for larger area of the brain, use tile-scanning with a 0.6 zoom factor and 10% overlap for automated reconstruction.
If acquiring z-stack images use 1-4um spacing. Use stack projection in ImageJ and exclude approximately 10% of the section's surface.
Making sure to use the same settings across all images for the same experiment, use ImageJ to process the images.