Imaging- Confocal
daniel.dautan daniel, Per Svenningsson
Published: 2024-07-13 DOI: 10.17504/protocols.io.j8nlk88y1l5r/v1
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.
Steps
1.
Using the confocal microscope, capture images at 10x magnification using a resolution of either 1024x1024 or 2048x2048.
2.
If needed for larger area of the brain, use tile-scanning with a 0.6 zoom factor and 10% overlap for automated reconstruction.
3.
If acquiring z-stack images use 1-4um spacing. Use stack projection in ImageJ and exclude approximately 10% of the section's surface.
4.
Making sure to use the same settings across all images for the same experiment, use ImageJ to process the images.
