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Microscope slide preparation methods - mold pollen bioaerosols - for indoor air quality analysis - Mold Testing Laboratory SOP MoldAPedia © Microscope slide preparation methods - mold pollen bioaerosols - for indoor air quality analysis - Mold Testing Laboratory Standard Operating Procedures SOPs Recipes and Formulas for microscope slide preparation mounting media Names and Sources for microscope slide mount preparation chemicals, media, and equipment, especially for aerobiologists and forensic particle examination

This is a collection of aerobiology and mold testing lab standard operating procedures - SOP's and slide preparation recipes useful for identification of mold, pollen, animal dander, skin cells, mite fecals, mouse dander, and other airborne allergens and bioaerosols. Comments and corrections are requested. © Copyright 2009 Daniel Friedman, All Rights Reserved. Information Accuracy & Bias Pledge is at below-left. Use links at the left of each page to navigate this document or to view other topics at this website. Green links show where you are in our document or website.

Slide Preparation Procedures & Guidelines

• Lab procedures: Cargille meltmountTM slides for rapid field or laboratory use, how to prepare
• Lab procedures: Microscopy - Microscope sources, different types of microscopes, microscope techniques and laboratory procedures for indoor air quality, toxic mold identification, & bioaerosol investigations
• Lab procedures: Slide preparation - tips on mold lab procedures, slide preparation recipes are provided beginning just below.

Microscope Slide Stain and Media Recipes

How to Prepare Permanent Microscope Slide Mounts

Here is a list of some candidates. WARNING: some of these involve dangerous chemicals, possibly carcinogenic (cancer-causing) and should be used only with expert advice..

• Canada Balsam - traditional, excellent slide mounting media, generally not available. See Cargille's "Meltmount" alternative.
• Clear nail polish - experimental -DF
• Damar Varnish - see Brinkworth and Smith article below
• Fructose Larry Legg's recipe
• Glycerin or glycerine jelly. DF note: we've been disappointed that even after adding phenol or other preservatives some of our glycerine jelly slides become cloudy or even opaque with age. We no longer recommend this mountant for permanent slide mounts.
• Gum Arabic (Acacia) or similar water soluble gums
• Hyrax / Dirax high refractive index mountants - Northern Biological Supplies
• Lactic acid combined with phenol (carbolic acid), polyvinyl alcohol, glycerine, water. (Not for caliciferous materials - you'll find that materials like drywall dust, gypsum, some plaster contents, will dissolve and then recrystalize in this media)
• Loctite glass bond - local auto supply stores, needs UV light to cure - experimental, Brinkworth and Smith [DF note: We have had good results using this slide preparation media for permanent mounts of certain particles; we need to research the refractive index but in general that's not been a problem with paint samples nor with fungal spores or other common indoor building dust particles. We use a halogen lamp to generate the UV needed to cure the mountant.
• Meltmount - see below
• Mowiol - available from Burkard as a replacement for Gelvatol - data to be added below
• NBS Bioseal Two
• Numount - from Brunel microscopes & Northern Biological Supplies
• Turtox - no longer available, formula sought. see Howey's article
• Water soluble gums - gum Arabic (Acacia)

How to Use Gelvatol to Prepare Microscope Slides - A Detailed Step by Step Procedure

Gelvatol may be used as a permanent slide mounting medium for mold spore or pollen slides. Gelvatol has been replaced by Mowiol from Burkard - see Mowiol below.

• 100 ml H2O (I would use distilled water)
• 35g Gelvatol (or Mowiol from Burkard)
• Polyvinyl alcohol (from Fisher Scientific, or other sources)

Heat to dissolve, not to much. Then add:

• 50 ml Glycerine (Glycerol, Fisher Cat. No. ?)
• 2g Phenol (as a preservative if needed)(Phenol smells a lot and it is no good for health, it gets yellow with time, I do not use it)

Add

• ACID fuchsin (as a colorant for SPORES if needed - Fisher Cat. No. AC22790-0250) Just use a few crystals, mix and wait. Add more if wanted, do not use too much as it will hide the structures.

OR BASIC fuchsin (as a colorant for POLLEN if needed - Fisher Cat. No. NC9653986)

OR no colorant

Keep closed tight because it gets harder as it gets in contact with air.

Source: Burkard company; I was told that they call the "polyvinyl alcohol" Gelvatol, available from Burkard.

...

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• Thanks to Alan Carson and Bob Dunlop, Carson Dunlop, Associates, Toronto, for permission to use illustrations from their publication, The Illustrated Home which illustrates construction details and building components. Carson Dunlop provides home inspection education, publications, report writing materials, and home inspection services. Alan Carson is a past president of ASHI, the American Society of Home Inspectors.
• Meltmount for Microscopic Slide Preparations: This product, available from Cargille Laboratories, is a contemporary, non-toxic replacement for Canada Balsam and has the same refractive index. Instructions for preparing slides using MeltMount are available from Cargille. I use this product in the lab for permanent slide mounts.
• Mowiol: his replacement for Gelvatol is provided by Burkard. I'll add more information here when I can find it.
• The Stains File stainsfile.info/StainsFile/jindex.html This is the most extensive list We have found of slide preparation stains. However it's quite shy on stains for pollen and mold spores. Above are listed stains and recipes of particular interest to aerobiologists and for which I have detail or alternatives beyond the information at the Stains File.
• Burkard Co. : We were told that Burkard's term for "polyvinyl alcohol" is Gelvatol - vailable from Burkard.
• Some microscope slide mounting technique articles
• Sealing Glycol Jelly (Glycerine) Mounts Between Two Cover Glasses William Ells
• Slide Mounting Made Easy Brinkworth and Smith
• Un mounting specimens from Balsam or Glycerine Jelly Slides Golden and Ellington -[SORRY this link is dead - contact Daniel Friedman for discussion about how to "un-mount" microscope specimens which have been mounted using Balsam or Glycerine Jelly. While careful heating will permit disassembly of a microscope slide prepared with these media, care may be needed to avoid losing particles during the removal of the slide cover-slip.]
• Permanent Slides, Pros/Cons Richard Howe

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• Lab procedures: Cargille meltmountTM slides for rapid field or laboratory use, how to prepare
• Lab procedures: Microscopy - Microscope techniques and laboratory procedures for indoor air quality, toxic mold identification, & bioaerosol investigations
• Lab procedures: Slide preparation - tips on mold lab procedures?
• Fisher Scientific lab supplies
• ICN Biomedicals biomedical products, books, supplies and MSDS information

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