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Intro to Histo- Electron Microscope Flashcards

1
Q

What are 7 types of Dyes for Electron Microscope?

A
  1. Sudan Stains
  2. Osmium Tetroxide Stain
  3. Periodic Acid Schiff (PAS) stain
  4. Alcian blue stain
  5. Trichrome Stain
  6. Van Gieson Stain
  7. Giemsa stain
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2
Q

What are 2 types of Metal impregnation techniques?

A
  1. Silver Stain
  • Reticular fibers (Silver salts)
    • Type 2 Collagen
  1. Golgi’s method
    * For Nervous tissue
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3
Q

What are 3 main Heavy Metal Ion Stain?

A
  1. Lead citrate
  2. Uranyl acetate
  3. Osmium tetroxide
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4
Q

What is Oil red O Stain?

A
  1. Sudan IV stain similiar to Oil Red O stain
  2. Used to detect Fat Emboli in Lungs (after a fracture of the lower leg)
    * Identifies tumors, lipomas, and liposarcomas
  3. Stains
  • Lipids: RED
  • Nuclei: BLUE/BLACK
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5
Q

What is Sudan Black Stain?

A
  1. Stains
    * Lipids: BLACK
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6
Q

What is Osmium Tetroxide Stain?

A
  1. Stains
  • Lipids: BLACKish-Brown
  • Myelin: Black
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7
Q

What is Periodic acid-Schiff (PAS)?

A
  1. Used for Carbohydrates macromolecules (polysaccrides)
  2. Stains
    * Magenta
  3. Carbohydrates
  • Glycogen- Muscle cells (Hepatocytes)
  • Glycoprotein (thyroid tissue)
    • Ex: Mucin
      • Glycocalyx
      • Basal lamina
      • Lysosomal enzymes (phagocytosis)
  • Proteoglycan (CT and ECM)
    • Form gels
      • Lubricates joints
      • Holds cells and tissue
      • Flexiblity of cartilage
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8
Q

What is Alcian blue?

A
  1. Stains
    * Acid Mucin: BLUE
  2. Does not stain neutral mucin (alkaline)
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9
Q

What does Alcian blue/ PAS combination stain?

A
  1. Stains
    * Apical neutral/acid mucin droplets of intestinal goblet: BLUE-PURPLE
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10
Q

How does PAS staining help with diagnosing?

A
  1. Glycogen storage disease
  2. Adenocarcinomas and Cystic Fibrosis
    * Increased mucin production
  3. Paget disease
    * Of the breast and nipple
  4. Whipple’s disease
  • Infectious disease that first affect GI tract and causes malabsorption
  • PAS stains: MACROPHAGES
  1. Mycosis fungoides
    * Aggregates of PAS positive lymphocytes in epidermis
  2. Erythroleukemia
  • A leukemia of immature red blood cells
  • Stain: Bright Fuchsia (Purple-red)
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11
Q

What are 3 main CT stains?

A
  1. Trichrome stain
  2. Elastic fiber stain
    * Van Gieson
  3. Reticular fiber stain
    * Silver salt/stain
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12
Q

What is Trichrome Stain?

A
  1. Stains Collagen
  2. Useful for Liver Cirrhosis
    * Produces large amounts of collagen/scar tissue
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13
Q

What is Mallory Trichrome Stain?

A
  1. Used on CT
    * Visualize collagen and reticular fibers
  2. Stains
  • Collagen and Reticular fibers: BLUE
  • Smooth muscle and Nuclei: RED
  • Elastic fibers: PINK
  • Red Blood Cells: ORANGE
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14
Q

What is Masson’s Trichrome?

A
  1. Helpful for differentiating Cellular from Extracellular Items
  2. Stains
  • Collagen fibers: GREEN
  • Smooth Muscle: RED
  • Cytoplasm: PINK to RED
  • Nuclei: BLACK
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15
Q

What is Elastic Fiber Stain? (Van Gieson)

A
  1. Differentiate between Collagen and Smooth Muscle
    * Tumors
  2. Stains:
  • Elastic Fibers: Black/Purple-black
  • Collagen: RED
  • Cytoplasm, muscle, RBC: YELLOW
  • Nuclei: BLACK/BLUE-BLACK
  • Cartilage: PINK
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16
Q

What are 2 types of Metal Impregnation Technique?

A
  1. Reticular Fiber Stains
    * Silver salts
  2. Golgi Silver Impregnation Technique
17
Q

What are Reticular fibers stain in Metal Impregnation technique?

A
  1. Stains
    * Reticular Fibers: BLACK
  2. Reticular fibers (reticulin)
  • Look like Fine Black Threads
  • Form networks
    • Support cellular components of Liver and Lymphoid organs
18
Q

What is the Golgi’s method?

A
  1. Most common for Nervous Tissue
  2. Stains
    * Neurons, Glia, and Vascular System: BLACK on a YELLOW/ORANGE background

*Image shows Pyramidal and Purkinje Cells

19
Q

What is Giemsa Stain?

A
  1. Used for BLOOD CELLS and SMEAR CELLS
    * Bone Marrow
  2. Stains:
  • Nuclei: BLUE to VIOLET and background pale BLUE
  • Erythrocytes: PALE PINK
  • Platelets and Leukocytes: BLUE
  1. Diagnosis for:
  • Malaria
  • Babesiosis
  • Sleeping Sickness
  • Schistosomiasis
20
Q

What is Heavy Metal Ion Stain?

A
  1. Used
  • Transmission Electron Microscopy
    • Subcellular structures
  1. Stain
    * Tissue: BLACK
21
Q

What is the steps of Tissue Preparation for TEM?

A
  1. Size of sample
    * 1 mm3
  2. Routine Fixants
  • Glutaraldehyde
    • Perserve Protein constituent by cross-linking
  • Osmium Tetroxide
    • Perserve membrane lipids (phospholipids)
  1. Embedding
    * Epoxy Resin (Light microscope uses Wax)
  2. Sectioning
  • 40-90 nm
  • Uses ultramicrotome either glass or diamond knife
  1. Mounting
    * On Wire Mesh (grid)
22
Q

What is Tissue Preparation for SEM?

A
  1. Same except after dehydration, spray coated with thin layer of heavy metal (usually gold)
  2. Staining
  • Uranyl acetate: membrane structures and structures with nucleic acid
  • Lead Citrate: Binds to RNA-containing structures
23
Q

What is Transmission Electron Microscope (TEM)?

A
  1. Electrons transmitted through Specimen
  2. Resolution: 3 nm
  3. Observe: Organelles and ECM
  4. Imaging
  • Bright, white areas
    • Electron Lucent
  • Black, dark areas
    • Electron dense
24
Q

What is Scanning Electron Microscope (SEM)?

A
  1. Focused electron beam
    * DOES NOT PASS THROUGH SPECIMEN
  2. Image
    * Scans only show the surface view
25
Q

What is an Artifact?

A
  1. Distortion of the tissue
  • Ruins results
  • Shrinkage, wrinkles, spaces, precipitates from the stain
26
Q

What is Autoradiography?

A
  1. X-ray to localize newly synthesized macromolecules
  2. Specimen itself is the source of the radiation
27
Q

What is Classical X-rays?

A
  1. Specimen placed between source of radiation and the film
  2. Absorption and scattering of radiation
28
Q

What is Fibroblastic cells?

A
  1. Bipolar/Multipolor
  2. Elongated shape
  3. Grow attached to a substrate
29
Q

What is Epithelial-like cells?

A
  1. Polygonal
  2. Grow attached to substrate in discrete patches
30
Q

What is Lymphoblast-like cells?

A
  1. Spherical
  2. Grown in suspension without attaching to a surface
31
Q

What is Peroxidase?

A
  1. Promotes oxidation of substrates
    * Transfer of hydrogen ion to hydrogen peroxide
32
Q

What is Phosphatase?

A
  1. Removal of phosphate group from macromolecules
33
Q
A
  1. Kidney tubule treated histochemically
    * Demonstrates Alkaline phosphastase
34
Q
A
  1. TEM image of Kidney cell
    * Acid phosphatase
  2. Dark areas
    * Lead phosphate

Histology (11 decks)

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