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Morphology Service Laboratory

First developed as a practical research tool in the late 1940s, electron microscopy has emerged as a valuable approach for understanding how cell structure and function are interrelated. Transmission and scanning electron microscopy can provide investigators with information not obtainable by any other means.

The electron microscopy facility at the Virginia-Maryland Regional College of Veterinary Medicine is equipped with instrumentation for ultrastructural analysis of biological and non-biological materials and to provide investigators with data concerning specimen morphology. The Morphology Laboratory complies with Good Laboratory Practice standards.

Facilities are available for transmission and scanning electron microscopy, semi- and ultra thin-sectioning, light microscopy, computerized morphometric and three-dimensional reconstruction studies of electron micrographs, gross specimen photography, and some photographic processing.

Clinical investigations as well as applied and basic research are encouraged.

Instrumentation   |   Technical Expertise   |   Using the Lab      
Examples of Supported Research Projects        


A. Instrumentation

  1. General Equipment
    1. Balances
      • Fisher Scientific Allied (Model 8215A)
        Top loading electronic balance with capacity weighing range of 0 to 1500 g.
      • Mettler PM600
        Top loading electronic balance with capacity range of 0 to 610 g.
    2. Centrifuges
      • Beckman Microfuge 12
        Non-refrigerated tabletop centrifuge with a maximum speed of 12,500 rpm and a 15-minute timer.
      • IEC Clinical Centrifuge
        Centrifuge for general purpose use.
    3. Computer Resources
      • A number of personal computers are dedicated to instrument operation and image acquisitions for both light and Transmission Electron Microscopy.
      • Printers:
        - Hewlett Packard LaserJet Series 4
        - Hewlett Packard Color LaserJet 1600
    4. Fume Hoods
      • Captair Ductless Portable Fume Hood
        Eliminates both gases and fumes by filtration.
      • Kewaunee Airflow Fume Hood
        For use with toxic chemicals.
    5. Corning Stirring/Hotplates
      • For preparation of fixatives, reagents, and solutions.
    6. Microscopes - General
      • Leitz Dialux with Sony CCD camera
      • Nikon Optiphot SMZ-10
        High resolution zoom (0.66 to 4X magnification) Stereo Photo microscope with a UW 20X ocular and diascopic illumination stand as well as the Nikon MKII Fiber Optic light source.
      • Olympus BH-2 with PM-10AD photomicrographic system
      • Olympus CH-2
        10X ocular with 4X, 10X, 40X objectives plus a 100X oil immersion objective.
      • Vanox Olympus AH-2 with digital camera attachment and Optronics imaging archiving system
    7. Microwave Oven
      • Precision Pulsed Laboratory Microwave Oven with accurate temperature control, 3 timer modes, 5 mixing tubes, and adjustable temperature probe.
    8. Mixers
      • Orbitron Model II Rotator
      • Yamato Lab-Stirrer (Model LR-51B)
      • Vortex-genie
        Automatic "start and stop" operation with variable speed control to regulate its mixing action.
    9. Advanced Micro-Osmometer (Model 3MO)
      • Instrument used to measure the osmolarity of solutions.
    10. Ovens
      • Fisher Isotemp Vacuum oven Model 281 and 281A
      • Thermolyne oven/incubator
      • Techne Dri-block DB-3
    11. Fisher Acumet model 610A pH Meter
      • With compensation circuit for effects of solution temperature.
    12. Polaroid MP4 Land Camera and Copy Stand
      • Adaptable to both 35 mm and Polaroid films, the Polaroid MP4 Land Camera is an unusually versatile photographic unit; included in its many applications are photomicrography, photomacrography, slide making, small object and gross specimen photography.
    13. Scanners
      • Epson Perfection 636
      • Polaroid Sprint Scan 45
    14. Ultrasonic Cleaner
      • Branson Ultrasonic Cleaner
        Effective for mixing, dispersion, dissolution, and cleaning most materials; ineffective for cleaning soft or porous materials.
      • DuPont Diamond knife cleaner.
    15. Ladd Vacuum Desicator
      • To provide a dry, clean vacuum chamber for Electron Microscope Film and specimen storage.
    16. Water Bath (Triangle Biomedical Sciences)
  2. Major Equipment
    1. Ladd Critical Point Dryer
      • Rapidly and reliably dehydrates biological specimens for scanning electron microscopy while preserving the structure of the tissue by eliminating surface tension stresses.
    2. Electron Microscopes and Accessories
      • Phillips 505 Scanning Electron Microscope
        Bridging the magnification and resolution gap between the light and transmission electron microscope (TEM), the scanning electron microscope (SEM) can easily produce surface images over a broad range of magnification (20X to 80,000X) with a resolution of 60 Å. Its tremendous depth of field enables the SEM to produce vivid three dimensional images of a variety of samples.
      • Zeiss 10CA Transmission Electron Microscope is equipped with AMT Advantage GR/HR-B CCD Camera System (digital imaging system).
    3. SPI Module Sputter Coater
      • A cold process whereby metal atoms are liberated from a target by ion impact; the sputter coater provides fast, fine grained deposition of conductive films for SEM analysis; it has an integral quartz crystal thickness monitor for accurate deposition of precious metals (gold).
  3. Darkroom Equipment
    1. Dryers
      • Negative Dryer
        Designed for drying sheet film or projection slides, it is equipped with an interior chamber 9 1/2" x 9 1/2" x 9 1/2" and a 60 minute timer.
    2. Paper Cutters
      • RotaTrim Paper Cutter
        Rotary paper cutter with automatic clamp for holding photographs in position for trimming.
    3. Premier Paper Safe
      • Light, tight paper storage container designed to save time and prevent costly spoilage in the darkroom.
    4. Arkay Loadmaster Print Washer
      • Stainless steel rotating drum type washer which accommodates approximately 75 8" x 10" prints.
    5. Processors
      • Arkay Gas Burst Agitation Timer/Tank
        Nitrogen burst timing for uniform processing of electron image sheet film.
    6. Timers
      • Time-O-Lite
  4. Sectioning Equipment
    1. Diamond Knives
      • Standard
      • Cryoultramicrotomy
        Gem quality diamond knives are used for cutting flawless silver sections for ultra microtomy and cryomicrotomy. Diamond knives are to be used by qualified personnel only.
    2. Hotplates
      • Thermolyne
      • Corning Ceramic Top Hotplate
        Thermostatically controlled for constant temperature preparation and staining of thick sections for light microscopy.
    3. KnifeMakers
      • LKB 7800
        For the safe and reproducible production of glass knives of consistently high quality for sectioning electron microscopy samples.
      • LKB2078 Histo-KnifeMaker
        For the production of high quality Ralph knives with 25 or 38 mm cutting edge length.
    4. Microtomes and Accessories
      • Ultramicrotomes
        Precision instruments designed for consistent high quality ultrathin sections for electron microscopy to semi-thin sections for light microscopy; the microtomes include a continuous feed range from 0 nm to 2.5 µm utilizing adjustable cutting speeds for 0.1 to 50 mm/sec.

        - **Reichert-Jung Ultracut E
        - **Leica Ultracut UCT
        - LKB Ultratome IV
        - LKB Ultratome Nova
        - RMC MT6000-XL

        **The Reichert-Jung Ultracut E and the Leica Ultracut UCT ultramicrotomes are reserved for use by Morphology Laboratory personnel only. Investigators are permitted to use the LKB and RMC ultramicrotomes.
      • Microtomes for Light Microscopy:
        - LKB 2218 Historange Microtome
        Microprocessor-controlled microtome designed to cut sections for light microscopy with precision, accuracy, and speed.
    5. Microscopes (see General Equipment)

B. Technical Expertise

  • Processing of biological and non-biological materials for transmission, scanning, and light microscopy.
  • Semi-thin and/or ultra-thin ultramicrotomy.
  • Operation and routine maintenance of the transmission and scanning microscopes and ancillary equipment.
  • Photography - developing TEM film.
  • Preparation of buffers, solutions, and resins.
  • EM methodology and tissue preparation to meet individual research objectives.
  • Basic instruction and training of faculty, staff and graduate students for the independent research use of the ultrastructural laboratory equipment and facilities.
  • Ultrastructural Pathology
    Detailed ultrastructural information obtained from TEM evaluation aids surgical pathologist/clinicians to resolve differential diagnosis or decide that an exact diagnosis is not possible, e.g., examination for viruses, lipid storage disease, parasite identification, tumor identification.
  • Positive and/or negative staining of tissues, cell cultures, and viruses.
  • Metal evaporative techniques utilizing sputter coater.
  • Critical Point drying.
  • Electronic image recording for teaching and research.

C. Using the Morphology Service Laboratory

Sample preparation and use of the equipment are available on a fee basis to all University faculty, their graduate students, and members of the scientific and health care community. Charges for services should be discussed with the lab supervisor, Kathy Lowe, at 1-4811, or the faculty coordinator, Dr. Tom Caceci, at 1-7178.


D. Examples of Supported Research Projects

  • Ultrastructural analysis of normal tissue; drug-induced changes and natural and traumatic pathology of connective tissue, blood and lymph, skeletal muscle, nervous, heart and vascular, lymphatic systems, skin and skin tumors; integument of oral cavity and tongue, glandular, digestive, respiratory, urinary, and reproductive systems, as well as melon and baleen of whales. Studies include selected tissues from cetacean, equine, sheep, chicken, Japanese newt, parasitic and marine invertebrates, marine mammals, dogs, laboratory animals, fish, cell culture, and humans.
  • Morphological differentiation of neurotoxic and cytotoxic effects of chemicals on neuroblastoma cells.
  • Genomic characterization of Felix-01 Bacteriophage and its use as a biological Salmonellacidal treatment for poultry skin.
  • Diagnostic and contract service.
  • Examination of vascular changes as a result of prophylactic vaccination of P. haemolyticae.
  • Environmental degradation of non-wovens (SEM fabric decomposition study).
  • Identification and purification of fimbriae (B. fragilis).
  • Ischemic reperfusion injury in small intestine of the horse.
  • Distention in equine jejunum.
  • Treatment of small intestine injury with Carolina rinse.
  • Effects of a recirculating aquaculture system on gill morphology of hybrid striped bass.
  • Mucosal surface of the intestinal tract of the Nile tilapia, Oreochromis niloticus.
  • Biotriboloby: friction wear and lubrication on bovine cartilage.
  • The role of feline leukemia virus in the pathogenesis of feline osteochondromatosis.
  • Primary lens luxation in Shar-pei.
  • Characterization of DNA and protein utilizing immunocytochemistry on Arabidopsis (little weed).
  • Mitochondrial dysplasia/myopathy in equine muscle.
  • Anatomy, histology and histochemistry of tilapias.
    1. Ultrastructure and development of the tilapia liver.
    2. Scanning and ultrastructure of the tilapia stomach.
    3. Ultrastructure of the tilapia gill.
    4. Ultrastructure of the tilapia anterior and posterior kidney.
    5. Ultrastructure of gentamicin toxicity in the tilapia kidney.
    6. Ultrastructure and development of the tilapia intestine.
  • Examination of various stages of tilapia development.
  • Anatomy and histology in development of the digestive tract in turkeys.
  • Examination of cancer cells treated with various drugs.
  • Examination of heart stents' structures.
  • Nature and development of Teleost rodlet cells.
  • Identification of viral particles in lip fibromas of angelfish.
  • Characterization of rodlet cells in angelfish.
  • Ultrastructure of the tilapia rodlet cell development.
  • Ultrastructure of the hybrid striped bass gill.
Ultrastructural
  • Examination of myoendocrine cells and coronary microvessels of camel (Camelus dromedarius) heart.
  • Comparison of intestinal flora after alteration of diets in laboratory rats.
  • Changes in axonal morphology of human trigeminal nerves following traumatic injury.
  • Neuropathology of rats exposed to acrylamide, 2, 5- hexanedione, and organophosphates.
  • Examination of the digestive tract and gall bladder of the Black Mollie (Poecilia spp.), a hybrid teleost.
  • Study of the development of the equine embryonic capsule and loss of the zone pellucida.
  • Cell Culture Studies - Characterization of:
    1. Equine endometrial cells from primary culture with a comparison to endometrial tissue samples from the same animal.
    2. SY5Y human neuroblastoma cells.
    3. Equine lamellar and equine endothelial cells from primary culture.
    4. SP1K bottlenose dolphin kidney cells and subsequent characterization of cytotoxicity in cells exposed to #1 fuel oil.
    5. CCD33Co human colonic cells with subsequent cytotoxicity studies.
    6. Chicken brain reaggregates (CBR) and subsequent characterization of the cytotoxicity of organophosphates on CBRs.
    7. THP-1 human monocyte cell lines.
    8. Demonstration of Polysaccharide capsule in Campylobacter jejuni using electron microscopy.
    9. LVS monolayers
  • Changes in sperm in the Shetland sheep dog.
  • Examination of bed bugs.
  • Characterization of carpenter ant foregut.
  • Characterization of soft tissue stenosis of the canine lumbosacral spine using intravenous contrast enhanced computer technology.
  • Characterization of intestinal adenocarcinoma with Paneth cell differentiation in a Virginia Opossum.
  • Diagnosis of bovine familiar convulsions and ataxia in an Aberdeen Angus calf.
  • Examination of spinal connective tissues from dogs with lumbosacral stenoses, using immunohistochemistry.
  • Examination of cream and cake samples
  • Characterization of wild type and mutant Drosophila muscle and brain.
Polymers
  • Multiple phase behavior in model silicone networks.
  • Highly hydrated polymer systems.
  • Organic/inorganic hybrids (ceramers).

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