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Microscope slide preparation method
How to prepare slides of
mold pollen bioaerosols - for indoor air quality analysis,Mold Testing Laboratory SOP
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Aerobiolgy & forensic microscopy lab procedures: how to prepare microscope slides for lab analysis. 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 suggestions are
invited. Readers should also see LAB PROCEDURES MICROSCOPE TECHNIQUES. More microscope slide procedures are described at PAINT LAB SAMPLE PREPARATION.
Green links show where you are. © Copyright 2013 InspectAPedia.com, All Rights Reserved. Author Daniel Friedman.
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.
Our photo (left) shows particle traces from two different air sampling cassettes prepared on microscope slides for further evaluation. Our study comparing the performance of these cassettes is found at AIR TEST SAMPLING CASSETTE STUDY.
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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
Recipe for Gelvatol Slide Mount Medium
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.
Frequently Asked Questions (FAQs)
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Technical Reviewers & References
Related Topics, found near the top of this page suggest articles closely related to this one.
- AIRBORNE PARTICLE ANALYSIS METHODS - Airborne Particle Calculations,
How to calculate airborne particle concentrations for mold, IAQ, allergens, or other forensic particle studies - Mold Test Laboratory SOPs
- MICROSCOPE SLIDE PREPARATION - Microscope slide preparation methods - mold pollen bioaerosols - for indoor air quality analysis - Mold Testing Laboratory SOP
- LAB PROCEDURES MICROSCOPE TECHNIQUES - Microscope techniques and laboratory procedures for indoor air quality, toxic mold identification, & bioaerosol investigations
- LAB & FIELD IAQ EQUIPMENT SOP - Forensic Microscopy, Particle Identification & IAQ Investigation Field Equipment Specifications, including physical properties (such as particle trace dimensions) and calibration data for air sampling and other test equipment
- MICROSCOPE DIGITAL PHOTOGRAPHY - how to obtain sharp photographs of views through the microscope using a digital camera
- TECHNICAL & LAB PROCEDURES - Technical Procedure Articles on Testing for Mold and other Indoor Air Quality IAQ Contaminants
- For most-recent microscope optical measurements: field width data and calibration see: Olympus-CH-2.xls & Polam Optics.xls Calibration is repeated no less than when an objective is removed/replaced or other microscope optical or
stage components are removed/replaced, or if a new technician is employed.
- For most-recent calculations of % of trace represented by n cross-scans see: Particle Counts.xls
- For automated calculations of particles or spores per cubic meter of air (spores/M3)
from raw airborne particle counts see: @SporeCountWork.xls
- For spore count worksheets see: sheet 2 of @SporeCountWork.xls
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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.
- Some microscope slide mounting technique articles
Books & Articles on Building & Environmental Inspection, Testing, Diagnosis, & Repair
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- Environmental Health & Investigation Bibliography - our technical library on indoor air quality inspection, testing, laboratory procedures, forensic microscopy, etc.
- Adkins and Adkins Dictionary of Roman Religion discusses Robigus, the Roman god of crop protection and the legendary progenitor of wheat rust fungus.
- 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
- US EPA - Mold Remediation in Schools and Commercial Building [Copy on file at /sickhouse/EPA_Mold_Remediation_in_Schools.pdf ] - US EPA
- US EPA - Una Breva Guia a Moho - Hongo [Copy on file as /sickhouse/EPA_Moho_Guia_sp.pdf - en Espanol
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