Processing of a Mouse Brain for Downstream Analysis

Abstract

Source: Wager-Miller, J. et al. Collection of Frozen Rodent Brain Regions for Downstream Analyses. J. Vis. Exp (2020)

This video demonstrates a detailed step-by-step protocol for preparing frozen mouse brain tissue sections for downstream analyses.

Protocol

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.

1. Storing brains

1. Quickly remove the brains from euthanized adult CD1 wildtype mice weighing approximately 30 g using a conventional approach and flash-freeze for 60 seconds in either liquid nitrogen or isopentane pre-chilled with dry ice.
2. Store frozen brains at -80 °C in aluminum foil or conical tubes until use.

2. Preparing the brain matrix

1. Twenty-four hours before dissecting tissue, place a cl....

Results

Figure 1: Preparing frozen work surfaces. (A) A brain block is placed on a stack of gel packs within an insulated box and is then sandwiched between two additional gel packs. Aluminum foil is packed at each end of the block to facilitate cooling .......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
0.5 mm Mouse coronal brain matrice	Braintree Scientific	BS-SS 505C	Cutting block
0.5 mm Rat coronal brain matrice	Braintree Scientific	BS-SS 705C	Cutting block
1.0 mm Biopsy Punch with plunger	Electron Microscopy Sciences	69031-01
1.5 mL microcentrifuge tubes	Dot Scientific	229443	For storing frozen ROIs
1.5 mm Biopsy Punch with plunger	Electron Microscopy Sciences	69031-02
2.0 mm Biopsy Punch with plunger	Electron Microscopy Sciences	69031-03
4-12% NuPage gel	Invitrogen	NPO323BOX	protein gradient gel
Bioanalyzer System	Agilent	2100	RNA analysis system
Dounce tissue grinder	Millipore Sigma	D8938	Glass tissue homogenizer
Fiber-Lite	Dolan-Jenner Industries Inc.	Model 180	Cool lamp
Glass plates	LabRepCo	11074010
Low profile blades	Sakura Finetek USA Inc.	4689