Mar 19, 2024
Mouse sample collection for metabolomics studies
- 1California Institute of Technology
Protocol Citation: Livia Hecke Morais 2024. Mouse sample collection for metabolomics studies. protocols.io https://dx.doi.org/10.17504/protocols.io.14egn3pkzl5d/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 13, 2024
Last Modified: May 31, 2024
Protocol Integer ID: 96675
Keywords: ASAPCRN
Funders Acknowledgement:
ASAP
Grant ID: ASAP-020495
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Abstract
Protocol used in the Mazmanian lab for collecting brain and gut tissues and plasma from mouse for metabolomics
Samples collection for metabolomics studies
Samples collection for metabolomics studies
At 4 months of age, mice were sacrificed by
decapitation
The brain was rapidly removed from the skull and placed in an ice-chilled stainless steel coronal
matrix
Brain tissue was sectioned in slices of approximately 1 mm
Substantia nigra, striatum, motor cortex(referred to as cortex) and caudal brainstem (referred to as brainstem) were dissected within three minutes using a brain atlas as reference1
Gut tissue and contents were dissected on an ice-chilled stainless steel dissection
tray
All tissue samples were weighed, stored in and stored at Precellys‱ 2 mL Soft Tissue Homogenizing Ceramic Beads tubes (Catalog number: 10011152), snap-frozen in dry ice and stored at -80ºC
until processing
Trunk blood was collected in BD Vacutainer‱ Hemogard‱ Closure Plastic K3-Edta Tubes (Catalog number: 15349700) and kept at room temperature before plasma separation
Plasma was separated by centrifugation at 2,500 x g for 10 minutes
Plasma was transferred to a pre-cooled collection vial Thermo Scientific Pierce 1.5 mL Capacity Microcentrifuge Tubes (Catalog number: 69715) and stored at -80ºC until processing
References:
1. Paxinos, G. and Franklin, K.B.J. (2001) The Mouse Brain in Stereotaxic Coordinates. 2nd Edition, Academic Press, San Diego.