Nov 22, 2023
4% Paraformaldehye in .1M PB preparation
- 1HHMI Janelia Research Campus
Protocol Citation: Monique Copeland 2023. 4% Paraformaldehye in .1M PB preparation. protocols.io https://dx.doi.org/10.17504/protocols.io.ewov1yr6kvr2/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: August 17, 2019
Last Modified: November 22, 2023
Protocol Integer ID: 26993
Keywords: perfusion solution, preparation
Funders Acknowledgements:
HHMI
Abstract
For perfusion solution.
Materials
MATERIALS
ParaformaldehydeCatalog #19200
Protocol materials
ParaformaldehydeCatalog #19200
Sodium Hydroxide, 10.0NThomas ScientificCatalog #C871W32
Corning™ Disposable Vacuum Filter/Storage Systems Fisher ScientificCatalog #09-761-5
Falcon Tube (50 mL)Fischer Scientific
Sodium Phosphate Dibasic Dihydrate (USP/FCC/EP/BP), Fisher ChemicalFisher ScientificCatalog #S472-500
Sodium Phosphate Monobasic Anhydrous (Colorless-to-White Crystals), Fisher BioReagentFisher ScientificCatalog #BP329-500
Paraformaldehyde prep
Add 400 mL of MilliQ water to a 1000 mL beaker
Equipment
Milli-Q® Integral Water Purification System for Ultrapure Water
NAME
filter
TYPE
Millipore Sigma
BRAND
ZRXQ010WW
SKU
LINK
Add stir bar in beaker and set on hot plate at 50 °C
Add 5 drops (500ul) of 10N NaOH (from 60ml dropper)
Sodium Hydroxide, 10.0NThomas ScientificCatalog #C871W32
(this makes the solution basic to help dissolve the paraformaldehyde)
Weigh 20 g of Paraformaldehyde and add to MilliQ water.
(Stir about 01:00:00 or until all parafromaldehyde is dissolved and solution is clear)
ParaformaldehydeCatalog #19200
Make 1 Mass Percent Phosphate buffer at PH 7.4
Prepare 1 Mass Percent Soduim Phosphate Dibasic Dihydrate (Store stock at Room temperature .)
Sodium Phosphate Dibasic Dihydrate (USP/FCC/EP/BP), Fisher ChemicalFisher ScientificCatalog #S472-500
Prepare 1 Mass Percent Sodium Phospate Monobasic Anyhydrous (store stock at Room temperature )
Sodium Phosphate Monobasic Anhydrous (Colorless-to-White Crystals), Fisher BioReagentFisher ScientificCatalog #BP329-500
To make 100 mL 1M PB at PH 7.4 add 19 mL Monobasic plus 81 mL Dibasic (store as stock at Room temperature )
Add 50 mL 1M PB PH 7.4 to paraformaldehyde solution.
Test PH and adjust solution to final desired pH using HCl or NaOH.
Top off to 500ml with MilliQ water.
Filter solution through .2um CA member filter.
Corning™ Disposable Vacuum Filter/Storage Systems Fisher ScientificCatalog #09-761-5
Lable container with '4% paraformaldehyed in .1M PB' and date. Store at 4 °C for up to 1 week.
(Can also be aliqouted into
Falcon Tube (50 mL)Fischer Scientific
and stored at -20 °C for 3 months.)