Apr 23, 2025
Version 2
Original
Aqueous (SBiP) Delipidation of a Whole Mouse Brain V.2
- Andrew Recknagel1,
- Kevin Cao2,
- Naveen Ouellette2,
- Molly Logsdon2,
- Judith Baka2,
- Jayaram Chandrashekar2,
- Rajvi Javeri2
- 1Janelia Research Campus;
- 2Allen Institute for Neural Dynamics
- Allen Institute for Neural Dynamics
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Abstract
Aqueous strategies for whole-brain delipidation involve lipid removal via phase separation and detergent washes. SBiP is an aqueous biphasic buffer that extracts lipids at polar-nonpolar solvent interfaces. The brain is then washed with the detergent-based B1n buffer, which further disrupts residual membranes, forming micelles that can be washed out. When paired with organic delipidation, these steps can return a solvent-shrunken brain to normal size in phosphate buffer, suitable for post-delipidation antibody labeling.
If performing this delipidation as part of the Whole Mouse Brain Delipidation, Immunolabeling, and Expansion Microscopy protocol, first delipidate the whole mouse brain with organic solvents using the THF and DCM Delipidation of a Whole Mouse Brain protocol.
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Create free accountSBiP Delipidation
5d 12h
Delipidate with aqueous SBiP solution
Start with a whole mouse brain perfused with 4% PFA, post-fixed, stored in PBS in a 20 mL vial.
Use a 20 mL vial for processing an adult mouse brain. All steps in this section are carried out on a carousel rotator 10 rpm, Room temperature .
Replace solution in vial with 20 mL SBiP for each of the following steps:
- SBiP for 03:00:00
- SBiP for 06:00:00
- SBiP Overnight
- SBiP for 24:00:00
- SBiP for 24:00:00
- SBiP for 24:00:00
- SBiP for 24:00:00
Note
It is important that the SBiP solution is mixed thoroughly during the aqueous steps in order to effectively delipidate the brain. When it is thoroughly mixed, the solution should look uniformly turbid at .
It is important that the SBiP solution is mixed thoroughly during the aqueous steps in order to effectively delipidate the brain. When it is thoroughly mixed, the solution should look uniformly turbid at Room temperature .
Safety information
2-Methyl-2-butanol and 2-propanol are corrosive and flammable. Perform the steps that involve these reagents under the fume hood. Wear a lab coat, safety goggles or glasses, and gloves.
2-Methyl-2-butanol and 2-propanol are corrosive and flammable. Perform the steps that involve these reagents under the fume hood. Wear a lab coat, safety goggles or glasses, and gloves.
4d 9h
Wash the brain with 20 mL B1n buffer at Room temperature Overnight .
Replace B1n buffer to complete ~24:00:00 wash.
1d
Wash the brain with 1X PBS, rotating at Room temperature for the following steps:
- PBS for 01:00:00
- PBS for 02:00:00 +
3h