Title: Standard Operating Procedure – IF staining on mouse brain tissue sections for VGLUT1/Homer1
Author: Rodrigo Sandoval Contreras, PhD and Nadia Haghbin, PhD
Date: 02/12/2025
Version: 1
Abstract: This SOP provides a standardized workflow for immunofluorescence staining of mouse brain tissue sections to detect the pre- and postsynaptic markers VGLUT1 and Homer1. It outlines the major steps of the procedure, including perfusion, fixation, tissue storage strategies (short-term vibratome sections and long-term cryopreservation), and sectioning by either vibratome or cryostat. The protocol describes preparation of both free-floating and slide-mounted sections, followed by antigen retrieval using sodium citrate buffer. Subsequent steps include blocking, primary and secondary antibody incubation, nuclear counterstaining, mounting, and confocal imaging. Together, these procedures support reliable visualization of presynaptic and postsynaptic markers in mouse brain tissue.
DOI: coming soon
Link to Full SOP: mIFvglut1h1-v1
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Version Notes
[DATE]: Version [#]
- Adjusted the After reward pause from 5s to 10s
- Reasoning is explained at the end of the SOP, but was based on fibre photometry recordings that demonstrated mice needed more time to baseline
- Included an additional formula for more data we can analyze: Premature Responses
- Added another method to run the stimulus duration probe
- Clarified details about the Inter Trial Interval
- Specifically, we made it clearer that the ITI is variable, rather than a constant duration between each trial