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MICROSCOPE SLIDE PREP - PERMANENT MOUNTS
  Required materials
  To use meltmount slides
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More Information

Photograph of procedure for preparing permanent slide mounts

Preparing Cargille meltmount™ slides for rapid field or laboratory use
InspectAPedia®  -      

  • Detailed, photo-illustrated procedure for preparing permanent microscope slide mounts
  • Questions & answers about making permanent microscope slide mounts

This lab procedure describes a method for preparing permanent microscope slide mounts using Cargille's MeltMount. We use this method when preparing particle samples which are to be retained indefinitely, such as for reference slides or where the samples may be required in environmental or mold litigation. - (Photographs are available -To Be Added or on request).

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Required materials for permanent mount microscope slide preparation using Cargille Meltmount

  • Cargille Meltmount - Cargille Laboratories www.cargille.com or from secondary suppliers such as www.2spi.com permanent slide mounting media (replacement for Canada Balsam and Aroclor) ND=1.539 @25degC.
  • Hot plate, Slides, Colored paper, Needle, dropper, or implement for mountant transfer to slide.
  • Detailed instructions available from Cargille regarding slide preparation using Meltmount. Remember to work in a well-ventilated area, to be careful of hot surfaces and chemicals, and to wear polyethylene gloves to avoid getting solvents or mountant on skin surfaces.

For in-lab use Cargille describes a procedure for sample preparation in which the sample is placed on the slide, the cover slip installed, and meltmount is introduced to the corners of the cover slip using a dropper.

Cargille also describes a method of pre-coating slides with mountant for subsequent use in the field.

Because the container of Meltmount, as received from Cargille takes some time to heat completely for in-lab use, and because I work with a variety of field sample materials which can be slow following this routine, I have adapted Cargille's "Pressure method," intended for field use, to a combination of lab and field work as I describe below.

  1. Cut out colored paper to approximately 2" x 4" and place a piece on the hot plate. The paper will be used to provide added contrast to enable easy view of the area of the slide which has been coated with mountant. Draw on the paper an outline of one of your slides and then mark within that outline the area where you will ultimately want to place a cover slip on the slide.

    This permits production of uniform slides and speed work at the microscope by placing each sample in the same area on the microscope stage. Prepare several of these colored slips so that you can replace them when soiled or wet.
  2. Heat the mountant to the proper working temperature. Heat surface to 60-70 C - the meltmount will be liquid but not smoking if it's at the easiest working temperature. I heat the meltmount in its container to a lower temperature
  3. Place the colored, marked paper on the hot plate.
  4. Place the slide on the colored paper over the outlined area.
  5. Pick up a droplet of Cargille permanent slide mountant with your implement of choice. I use a dental tool to pick up a droplet of mountant, let it solidify on the end of the tool (takes only a second), then I use the melted drop on the tool to "paint" the target area on the slide as described below. Too much mountant may produce a messy slide with a lot of excess mountant to be cleaned off later. Too little mountant may not adequately handle large particles, leaving a tilted cover slip.
  6. When the slide has been "painted" with mountant move it to another area of the hot plate where it should remain until the mountant has been heated to a level, uniform state. Meanwhile you can be working on coating subsequent slides.
  7. When the mountant is level on the slide remove the slide to a cool, not cold, surface and let the slide cool to room temperature.
  8. Label the slide with blank labels at this stage to make it easy later to quickly see which side has the mountant.
  9. Store prepared microscope slides flat, mountant upright. I use a plastic slide box.

To use previously prepared Cargille Meltmount slides in the field or lab.

Photograph of procedure for preparing permanent slide mounts
  1. Transfer the sample to the area of mountant either by scraping the sample on to the slide, transferring by tape, or other method.
  2. Heat the slide briefly over a match or alcohol lamp.
  3. Install the cover slip.
  4. Permit the slide to stabilize: Cargille recommends pressing the slide between waxed paper and weighted by a book overnight. When I used an alcohol lamp to heat the slide before installing the cover slip this was not necessary.
  5. Clean off any excess mountant with a razor blade [after the slide has cooled or stabilized] and/or an appropriate solvent - Cargille recommends xylene or toluene. I had some success also using hexane followed by isopropyl alcohol for final cleanup. [Caution: hexane is a toxic carcinogen.]

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Questions & answers about making permanent microscope slide mounts

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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.
  • 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

Allergies, Allergens, Allergy Testing in Buildings - References & Products

  • Air Conditioning System Blower Fans & Filters Cascading for Optimum Indoor Air Quality
  • Allergen Tests in Buildings advice about how to test, what to look for, in evaluating the level of dog, cat, or other animal allergens in a building
  • "IgG Food Allergy Testing by ELISA/EIA, What do they really tell us?" Sheryl B. Miller, MT (ASCP), PhD, Clinical Laboratory Director, Bastyr University Natural Health Clinic - ELISA testing accuracy: Here is an example of Miller's critique of ELISA http://www.betterhealthusa.com/public/282.cfm - Townsend Letter for Doctors and Patients
    The critique included in that article raises compelling questions about IgG testing assays, which prompts our interest in actually screening for the presence of high levels of particles that could carry allergens - dog dander or cat dander in the case at hand.
    http://www.tldp.com/issue/174/IgG%20Food%20Allergy.html contains similar criticism in another venue but interestingly by the same author, Sheryl Miller. Sheryl Miller, MT (ASCP), PhD, is an Immunologist and Associate Professor of Basic and Medical Sciences at Bastyr University in Bothell, Washington. She is also the Laboratory Director of the Bastyr Natural Health Clinic Laboratory.
  • Allergens: Testing for the level of exposure to animal allergens is discussed at http://www.animalhealthchannel.com/animalallergy/diagnosis.shtml (lab animal exposure study is interesting because it involves a higher exposure level in some cases
  • Allergens: WebMD discusses allergy tests for humans at webmd.com/allergies/allergy-tests
  • Animal Allergens: Dog, Cat, and Other Animal Dander - Cleanup & Prevention Information for Asthmatics and regarding Indoor Air Quality.
  • Atlas of Mold Related Illness Symptoms & Complaints - long list of both documented, studied mold related illness, and complaints ascribed to mold contamination or allergens in buildings
  • Cat Dander: how to inspect and test a building for past or current presence of cats, cat hair, cat dander, and cat allergens
  • Clinical Atlas of Mold Toxicity - An Online Description of Toxic, Pathogenic, Allergenic Fungi, Fungal Diseases
  • Fiberglass Insulation Contains Mold© 2005 comments about a field study in process, & more about health hazards from fiberglass insulation
  • Humidity: What indoor humidity should we maintain in order to avoid a mold problem?
  • Mold Action Guide detailed guide on finding, removing, and preventing indoor mold contamination
  • Odors, Odor Detection, Smells, & Gases how to find and identify sources of noxious or toxic odors and gases
  • Other environmental risks, Our much longer list: Asbestos, carbon monoxide, electromagnetic fields, etc.
  • Ozone: The Use of Ozone Indoors for Control of Odors and Mold Removal in Buildings: A Summary of Hazards and False Claims.
  • Pollen Allergens: identification, plant pollen and indoor air quality
  • Products to Reduce Mold & Allergy Problems to reduce indoor mold or allergen levels: air cleaners, air purifiers, dust mite covers, vacuum cleaners, crawl space vents
  • Recognizing Allergens: What various indoor allergens look like - identification photos to help identify pollen, dust mites, animal dander, toxic or allergenic mold - Common Mold and other Allergens, Irritants, Remedies & Advice
  • Rodent control issues, including dander, fecal, and urine contamination of Buildings and Building insulation are discussed at our
  • Sewage and Septic backup contamination in Buildings: inspection, testing, remediation, & references to expert sources
  • Action Guide: What to do about mold, mildew, and other indoor allergens
  • ...

Mold Contamination Testing, Cleanup, Prevention: references & products

  • GO TO the MOLD and INDOOR ENVIRONMENT INFORMATION CENTER for in-depth advice on avoiding testing for or cleaning up mold and other indoor environmental hazards, odors, gases, contaminantsThe Mold Information Center: What to Do About Mold in Buildings, When and How to Inspect for Mold, Clean Up Mold, or Avoid Mold Problems
  • Aerobiology, Building Science, Microscopy, & Laboratory References, an extensive technical bibliography
  • Allergens: what they look like in buildings
  • 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) - The Atlas of Clinical Fungi is also available on CD ROM
  • Atlas of Mold Related Illness: Index of Symptoms and health, physical, neurological, psychological, and other complaint which people suspect may be mold or building-related.
  • 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 recognizable, 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?
  • Clinical Mold References - Detailed bibliography of mold reference texts
  • "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
  • "Management of Powdery Mildew, Leveillula taurica, in Greenhouse Peppers," Ministry of Agriculture and Lands, British Columbia - Original source: www.agf.gov.bc.ca/cropprot/peppermildew.htm
  • Environmental Health & Investigation Bibliography - our own technical library on indoor air quality inspection, testing, laboratory procedures, forensic microscopy, etc.
  • Fiberglass: Mold in Fiberglass Insulation© 2005 comments about a field study in process, & more about health hazards from fiberglass insulation - DJF
  • 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.
  • Fungi, Identifying Filamentous, A Clinical Laboratory Handbook, Guy St-Germain, Richard Summerbell, Star Publishing, 1996, ISBN 0-89863-177-7 (English) (buy at Amazon)
  • Looking for Mold Procedure: what mold is often found where in buildings - simple technical presentation
  • Meruliporia: the house eating fungus or "poria"
  • Mold Action Guide: Step-by-Step Instructions, What to do about mold, mildew, and other indoor allergens
  • MOLD APPEARANCE - WHAT MOLD LOOKS LIKE Photos of what mold looks like in buildings
  • MOLD APPEARANCE - STUFF THAT IS NOT MOLD Photos of NOT-mold material that is sometimes mistaken for mold
  • MOLD ATLAS & PARTICLES INDEX, Pathogens, Allergens and Other Indoor Particles - Medical Health Effects of Mold (separate online document)
  • MOLD BY MICROSCOPE Mold under the microscope - photo identification of the most common indoor molds found in buildings
  • Mold FAQs Answers to Most Questions about Indoor Mold, Mold Related Illness, Mold Cleanup, Mold Prevention
  • US EPA: Mold Remediation in Schools and Commercial Building [Copy on file at /sickhouse/EPA_Mold_Remediation_in_Schools.pdf ] - US EPA
  • Mold spores in the Home - a Photo ID Library for detection and identification of mold allergens
  • Mold Test Kits - How to Collect and Send Your Own Mold Sample to our mold testing lab or to any mold lab you wish
  • Most Common Indoor Molds Found in Buildings, A Table of
  • Mycology, Fundamentals of Diagnostic, Fran Fisher, Norma B. Cook, W.B. Saunders Co. 1998, ISBN 0-7216-5006-6 (buy this book at Amazon)
  • Ozone Warnings - Use of Ozone as a "mold" remedy is ineffective and may be dangerous.
  • Rot concerns in buildings-some building mold such as Meruliporia incrassata "Poria" risks serious rot and hidden structural damage
  • US EPA: Una Breva Guia a Moho - Hongo [Copy on file as /sickhouse/EPA_Moho_Guia_sp.pdf - en Espanol

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.

  • Fiberglass building insulation and HVAC duct work insulation hazards
  • Sewage and Septic backup contamination in buildings: inspection, testing, remediation, & references to expert sources
  • Other environmental risks: Asbestos, carbon monoxide, electromagnetic fields, environmental illness, fiberglass, MCS - multiple chemical sensitivity, toxic gases, etc
  • Indoor Gas Sampling Plan for Residential Buildings lists a number of toxic indoor gases which we test for, depending on the building complaint and building conditions
  • 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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