Microscope slide preparation method
How to prepare slides of
mold pollen bioaerosols - for indoor air quality analysis,Mold Testing Laboratory SOP
MICROSCOPE SLIDE PREPARATION
- CONTENTS: 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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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.
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
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)
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, a product available from Burkard.
Reader Question: getting rid of air bubbles trapped in the Gelvatol?
(Apr 4, 2014) Anonymous said:
I've got a lot of tiny air bubbles trapped in the gelvatol I prepared. I left it for a while and remove them, but more bubbles form when touch with a glass rod. How can I get rid of them?
Anon. I would try heating the gelvatol - not hot enough to make it bubble or you'll have to start all over. Gently heat and watch it. When viscosity is pretty low, try tapping the container on a surface.
Also I find that one or two small bubbles in the mountant can be easily pushed to one side when I put on the cover slip. Or if there is not enough mountant under the cover slip I use a pipette to introduce a bit more mountant from one edge, later wiping that edge clean.
If more bubbles are forming when you push down on the cover slip I suspect something else is going on, possibly a chemical reaction with the substance being examined, or or you are pushing out air that was trapped in the mounted particles. For example, gypsum (drywall dust) in some indoor dust samples will froth many mountants, and flower or insect parts or large samples of fairly large or disparate particles will sometimes trap air that needs to be expelled.
Often after the cover slip is in place I will press down on it gently with the non-pinching end of a small pair of tweezers over which I've mounted a rubber dropper bulb. The bulb on tweezers gives me a dual -purpose tool since the rubber allows me to press gently without risking breaking the cover slip, and it also makes easier the slight adjustment of the cover slip position.
Finally, watch out for using slide mountant chemicals that react with some of the particles present. For excample using an acidic slide mountant with particles containing drywall dust will lead to bubble formation.
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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
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
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
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
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.]
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