Dec 27, 2023

Public workspaceFixative preparation for Baermann funnel extraction

DOI
dx.doi.org/10.17504/protocols.io.5qpvo31nbv4o/v1
  • 1The Ohio State University
  • OSU Soybean Pathology and Nematology
Casey A Schlenker
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DOI: dx.doi.org/10.17504/protocols.io.5qpvo31nbv4o/v1
Protocol Citation: Casey A Schlenker 2023. Fixative preparation for Baermann funnel extraction. protocols.io https://dx.doi.org/10.17504/protocols.io.5qpvo31nbv4o/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: December 27, 2023
Last Modified: February 27, 2024
Protocol Integer ID: 92764
Abstract
Fixative preparation for Baermann funnel extraction
Materials
Materials for fixative preparation:
  • Para-formaldehyde powder (PFA)
  • Phosphate-buffered saline (PBS) tablets
  • 1M NaOH
  • Glycerol
Fixative Preparation
Fixative Preparation
50m
50m
Weigh Amount1.6 g PFA powder

Dissolve in Amount20 mL of DI water

Incubate in water bath at Temperature60 °C to help dissolve PFA powder for Duration00:40:00

40m
Add Amount20 µL of 1 M NaOH to further help dissolve PFA powder

Place back in Temperature60 °C water bath for Duration00:10:00

10m
If still not dissolved, add Amount10 µL more of 1M NaOH

Pull out of water bath and cool to TemperatureRoom temperature

Add two tablets of PBS to a Amount100 mL volumetric flask

Mix Amount20 mL of each solution to get 4% formaldehyde

Add Amount400 µL of glycerol to fixative

Ensure Ph7.5

Place fixative in a hot water bath till Temperature60 °C