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
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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.
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
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.
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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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.
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.]
Books & Articles on Building & Environmental Inspection, Testing, Diagnosis, & Repair
Our recommended books about building & mechanical systems design, inspection, problem diagnosis, and repair, and about indoor environment and IAQ testing, diagnosis, and cleanup are at the InspectAPedia Bookstore. Also see our Book Reviews - InspectAPedia.
Kansas State University, department of plant pathology, extension plant pathology web page on wheat rust fungus: see http://www.oznet.ksu.edu/path-ext/factSheets/Wheat/Wheat%20Leaf%20Rust.asp
"A Brief Guide to Mold, Moisture, and Your Home",
U.S. Environmental Protection Agency US EPA - includes basic advice for building owners, occupants, and mold cleanup operations. See http://www.epa.gov/mold/moldguide.htm
Associations: Sick House, Sick Building, SBS - Air Quality, Government, Private Associations and Information Resources
Atlas of Clinical Fungi, 2nd Ed., GS deHoog, J Guarro, J Gene, & MJ Figueras, Centraalbureau voor Schimmelcultures, Universitat Rovira I Virgili, 2000, ISBN 90-70351-43-9 (you can buy this book at Amazon)
Atlas of Indoor Mold, Online Clinical Mold Atlas, Toxins, Pathogens, Allergens and Other Indoor Particles - Medical Health Effects of Mold (separate online document)
Black Mold that is Harmless Photos of recogniziable, usually harmless black mold on wood, bluestain, ceratocystis, ophistoma
Building Floods: quick steps after a building flood or plumbing leak can prevent costly mold contamination
Classes of Mold: what types of cosmetic, allergenic, or toxic mold are a problem? Can mold be cleaned-up successfully?
"A Brief Guide to Mold, Moisture, and Your Home", U.S. Environmental Protection Agency US EPA - includes basic advice for building owners, occupants, and mold cleanup operations. See http://www.epa.gov/mold/moldguide.htm
"Disease Prevention Program for Certain Vegetable Crops," David B. Langston, Jr., Extension Plant Pathologist - Vegetables, University of Georgia (PDF document) original source: www.reeis.usda.gov/web/crisprojectpages/209797.html
"Disease Prevention in Home Vegetable Gardens,"
Patricia Donald,
Department of Plant Microbiology and Pathology,
Lewis Jett
Department of Horticulture, University of Missouri Extension - extension.missouri.edu/publications/DisplayPub.aspx?P=G6202
Fifth Kingdom, Bryce Kendrick, ISBN13: 9781585100224, is available from the InspectAPedia online bookstore - we recommend the CD-ROM version of this book. This 3rd/edition is a compact but comprehensive encyclopedia of all things mycological. Every aspect of the fungi, from aflatoxin to zppspores, with an accessible blend of verve and wit. The 24 chapters are filled with up-to-date information of classification, yeast, lichens, spore dispersal, allergies, ecology, genetics, plant pathology, predatory fungi, biological control, mutualistic symbioses with animals and plants, fungi as food, food spoilage and mycotoxins.
OTHER IAQ ISSUES: How To Find and Address Other Indoor Air or Indoor Environment Contaminants Besides Mold
Mold or allergens may not be the only or even the main indoor environmental contaminant. Don't let media attention to mold
cause so much enviro-scare fear that other, possibly more urgent hazards go un-addressed.
Ozone Warnings - Use of Ozone as a "mold"
remedy is ineffective and may be dangerous.
Pet control - if you can't say goodbye to your bird, cat, dog, guinea pig, hamster, tropical fish, then limit the
areas they occupy and limit the airflow from that area to sleeping or other areas of the building, use allergenic
bedding, eliminate wall-to-wall carpeting, improve housecleaning including use of a HEPA-rated vacuum cleaner. For more details
see our article Dog, Cat, and Other Animal Dander - Information for Asthmatics and Indoor Air Quality
Rodents, Mice, Squirrel Control - I find high levels of mouse and rodent dander, fecal dust, and urine-contaminated dust in some buildings,
and high levels of these materials in building insulation in those locations. If you have a mouse problem, particularly if mice and their waste (fecals or urine) are contaminating
the building HVAC or building insulation, may need both steps to clean up or remove infected materials and steps to stop an ongoing
rodent problem. If squirrels are a problem, the cleanup needs to include closing off entry openings into the building. Get some
help from a licensed pest control expert.
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