Mouse sample collection for metabolomics studies
Livia Hecke Morais
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Abstract
Protocol used in the Mazmanian lab for collecting brain and gut tissues and plasma from mouse for metabolomics
Steps
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 VacutainerTM HemogardTM 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:
- Paxinos, G. and Franklin, K.B.J. (2001) The Mouse Brain in Stereotaxic Coordinates. 2nd Edition, Academic Press, San Diego.
