May 02, 2024
BrainSaw 50 mM pH 7.4 PB slicing buffer
- 1Sainsbury Wellcome Centre, UCL
Protocol Citation: Rob Campbell 2024. BrainSaw 50 mM pH 7.4 PB slicing buffer. protocols.io https://dx.doi.org/10.17504/protocols.io.q26g714r3gwz/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: May 02, 2024
Last Modified: May 02, 2024
Protocol Integer ID: 99107
Abstract
This protocol makes 2L of pH 7.4 phosphate buffer. This solution is used for serial section 2-photon imaging.
Materials
2 L GL45 bottle
magnetic stirrer
weighing scales
Protocol materials
Sodium phosphate monobasic monohydrateCatalog #S23120-1000.0
In 2 steps
Sodium phosphate dibasic heptahydrate MelfordCatalog #S23175-5000.0
In 2 steps
Safety warnings
Ensure the first solute is completely dissolved before adding the second. Not doing this will result in a small quantity of fine precipitate being formed.
Fill 2L flask with MQ water to the 2L notch. Place on magnetic stirrer and set to 1000 RPM.
Weigh 3.1 g of Sodium Phosphate Monobasic MonohydrateMelfordCatalog #S23120-1000.0 and pour into the flask.
Weigh out 20.8 g of Sodium phosphate dibasic heptahydrate MelfordCatalog #S23175-5000.0 but do not pour into flask until existing solute is dissolved.
Wait until Sodium Phosphate Monobasic MonohydrateMelfordCatalog #S23120-1000.0 has dissolved. Skipping this step will result in a small quantity of fine precipitate being formed.
5m
Pour Sodium phosphate dibasic heptahydrate MelfordCatalog #S23175-5000.0 weighed out above into flask. Screw on lid and shake once to ensure no solute is stuck around the rim of the flask. Return to stirring plate at 1000 RPM.
10m
Use PB once all solute is dissolved. Do not refrigerate: cold PB will form bubbles as it warms and this interferes with imaging.