Jun 13, 2023

Public workspaceTissue and blood collection

DOI
dx.doi.org/10.17504/protocols.io.bp2l69wmklqe/v1
  • 1Mitochondria and Inflammation in Neurodegenerative Diseases, DZNE, Tübingen-Germany;
  • 2Hertie Institute for Clinical Brain Research, University of Tübingen;
  • 3Neuropharmacology Laboratory, Oasi Research Institute-IRCCS, Troina, Italy;
  • 4Biomedical and Biotechnological Sciences, Pharmacology Section, University of Catania-Italy
carmela giachino
Icon indicating open access to content
QR code linking to this content
DOI: dx.doi.org/10.17504/protocols.io.bp2l69wmklqe/v1
Protocol Citation: michela.deleidi, Bianca Marchetti, Federico Bertoli, Carmela Giachino 2023. Tissue and blood collection. protocols.io https://dx.doi.org/10.17504/protocols.io.bp2l69wmklqe/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: March 31, 2023
Last Modified: June 13, 2023
Protocol Integer ID: 79800
Keywords: Tissue collection
Funders Acknowledgement:
ASAP-Aligning Science Across Parkinson’s
Grant ID: ASAP-000420
Abstract
This protocol details tissue collection from mice.
Attachments
Icon for k4dmb48if.docx
k4dmb48if.docx
13KB
Materials
Materials

  • 0.9% NaCl
  • 4% paraformaldehyde
  • phosphate buffer
  • EDTA-treated tubes
  • 15% sucrose solution
Tissue and blood collection
Tissue and blood collection
For immunohistochemistry, mice (5-6 mice/genotype/age-group/treatment) were anesthetized and transcardially perfused with 0.9% NaCl, followed by 4 % paraformaldehyde in phosphate buffer (pH 7.2 at 4°C), the brains and peripheral tissues carefully removed and post-fixed for 2-4 hrs, in 4% paraformaldehyde in phosphate buffer (pH 7.2) and placed in 15% sucrose in the solution of phosphate buffer overnight at 4°C.

Blood was collected into EDTA-treated tubes and serum was separated by centrifugation and stored at -20C.

Centrifigation
For gene expression and protein analysis, animals were sacrificed by cervical dislocation, the brains and peripheral tissues were quickly isolated and then stored at -80°C until assayed, the tails collected from each mouse were genotyped through PCR followed by electrophoretic analysis, as described in the manufacturer’s instructions (Jackson Laboratory).