May 08, 2024
BrainSaw agar stock solution
This protocol is a draft, published without a DOI.
- 1Sainsbury Wellcome Centre, UCL
Protocol Citation: Rob Campbell 2024. BrainSaw agar stock solution. protocols.io https://protocols.io/view/brainsaw-agar-stock-solution-ddcv22w6
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: May 08, 2024
Last Modified: May 08, 2024
Protocol Integer ID: 99445
Keywords: anatomy
Abstract
Making agar stock solution for serial sectioning imaging.
Materials
100 ml bottle
weighing scales
microwave for melting the agar
Protocol materials
Agarose multipurpose, molecular biology gradeMP BiomedicalsCatalog #11AGAP0100-CF
Step 4
Safety warnings
Hot agar can be painful but is not dangerous.
Before you start
Before you start
You will need at least 100 mL of 50 mM PB.
Protocol
NAME
BrainSaw 50 mM pH 7.4 PB slicing bufferCREATED BY
Rob Campbell
Make the stock
Make the stock
Rinse out the 100 ml bottle in which the stock is kept.
Fill the bottle with 100 mL of 50 mM PB to the notched fill line.
Weigh out 5 g of Agarose multipurpose, molecular biology gradeMP BiomedicalsCatalog #11AGAP0100-CF and pour into the bottle. Cap the bottle and shake it.
Using the stock
Using the stock
Do not heat the stock solution bottle. This concentrates it over time and it takes ages to heat. Instead, pour out the desired amount into a beaker and heat carefully in the microwave. Use low or medium power and stop as it begins to boil or even just before. The agar will look slightly less clear if you don't boil it, but this does not matter.
As a rule of thumb: melt just over 10 ml for a single brain. Just under 20 ml for 4 brains.