Intro to Histo- Electron Microscope Flashcards
1
Q
What are 7 types of Dyes for Electron Microscope?
A
- Sudan Stains
- Osmium Tetroxide Stain
- Periodic Acid Schiff (PAS) stain
- Alcian blue stain
- Trichrome Stain
- Van Gieson Stain
- Giemsa stain
2
Q
What are 2 types of Metal impregnation techniques?
A
- Silver Stain
- Reticular fibers (Silver salts)
- Type 2 Collagen
- Golgi’s method
* For Nervous tissue
3
Q
What are 3 main Heavy Metal Ion Stain?
A
- Lead citrate
- Uranyl acetate
- Osmium tetroxide
4
Q
What is Oil red O Stain?
A
- Sudan IV stain similiar to Oil Red O stain
- Used to detect Fat Emboli in Lungs (after a fracture of the lower leg)
* Identifies tumors, lipomas, and liposarcomas - Stains
- Lipids: RED
- Nuclei: BLUE/BLACK
5
Q
What is Sudan Black Stain?
A
- Stains
* Lipids: BLACK
6
Q
What is Osmium Tetroxide Stain?
A
- Stains
- Lipids: BLACKish-Brown
- Myelin: Black
7
Q
What is Periodic acid-Schiff (PAS)?
A
- Used for Carbohydrates macromolecules (polysaccrides)
- Stains
* Magenta - Carbohydrates
- Glycogen- Muscle cells (Hepatocytes)
- Glycoprotein (thyroid tissue)
- Ex: Mucin
- Glycocalyx
- Basal lamina
- Lysosomal enzymes (phagocytosis)
- Ex: Mucin
- Proteoglycan (CT and ECM)
- Form gels
- Lubricates joints
- Holds cells and tissue
- Flexiblity of cartilage
- Form gels
8
Q
What is Alcian blue?
A
- Stains
* Acid Mucin: BLUE - Does not stain neutral mucin (alkaline)
9
Q
What does Alcian blue/ PAS combination stain?
A
- Stains
* Apical neutral/acid mucin droplets of intestinal goblet: BLUE-PURPLE
10
Q
How does PAS staining help with diagnosing?
A
- Glycogen storage disease
- Adenocarcinomas and Cystic Fibrosis
* Increased mucin production - Paget disease
* Of the breast and nipple - Whipple’s disease
- Infectious disease that first affect GI tract and causes malabsorption
- PAS stains: MACROPHAGES
- Mycosis fungoides
* Aggregates of PAS positive lymphocytes in epidermis - Erythroleukemia
- A leukemia of immature red blood cells
- Stain: Bright Fuchsia (Purple-red)
11
Q
What are 3 main CT stains?
A
- Trichrome stain
- Elastic fiber stain
* Van Gieson - Reticular fiber stain
* Silver salt/stain
12
Q
What is Trichrome Stain?
A
- Stains Collagen
- Useful for Liver Cirrhosis
* Produces large amounts of collagen/scar tissue
13
Q
What is Mallory Trichrome Stain?
A
- Used on CT
* Visualize collagen and reticular fibers - Stains
- Collagen and Reticular fibers: BLUE
- Smooth muscle and Nuclei: RED
- Elastic fibers: PINK
- Red Blood Cells: ORANGE
14
Q
What is Masson’s Trichrome?
A
- Helpful for differentiating Cellular from Extracellular Items
- Stains
- Collagen fibers: GREEN
- Smooth Muscle: RED
- Cytoplasm: PINK to RED
- Nuclei: BLACK
15
Q
What is Elastic Fiber Stain? (Van Gieson)
A
- Differentiate between Collagen and Smooth Muscle
* Tumors - Stains:
- Elastic Fibers: Black/Purple-black
- Collagen: RED
- Cytoplasm, muscle, RBC: YELLOW
- Nuclei: BLACK/BLUE-BLACK
- Cartilage: PINK
16
Q
What are 2 types of Metal Impregnation Technique?
A
- Reticular Fiber Stains
* Silver salts - Golgi Silver Impregnation Technique
17
Q
What are Reticular fibers stain in Metal Impregnation technique?
A
- Stains
* Reticular Fibers: BLACK - 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
- Most common for Nervous Tissue
- 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
- Used for BLOOD CELLS and SMEAR CELLS
* Bone Marrow - Stains:
- Nuclei: BLUE to VIOLET and background pale BLUE
- Erythrocytes: PALE PINK
- Platelets and Leukocytes: BLUE
- Diagnosis for:
- Malaria
- Babesiosis
- Sleeping Sickness
- Schistosomiasis
20
Q
What is Heavy Metal Ion Stain?
A
- Used
- Transmission Electron Microscopy
- Subcellular structures
- Stain
* Tissue: BLACK
21
Q
What is the steps of Tissue Preparation for TEM?
A
- Size of sample
* 1 mm3 - Routine Fixants
- Glutaraldehyde
- Perserve Protein constituent by cross-linking
- Osmium Tetroxide
- Perserve membrane lipids (phospholipids)
- Embedding
* Epoxy Resin (Light microscope uses Wax) - Sectioning
- 40-90 nm
- Uses ultramicrotome either glass or diamond knife
- Mounting
* On Wire Mesh (grid)
22
Q
What is Tissue Preparation for SEM?
A
- Same except after dehydration, spray coated with thin layer of heavy metal (usually gold)
- 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
- Electrons transmitted through Specimen
- Resolution: 3 nm
- Observe: Organelles and ECM
- Imaging
- Bright, white areas
- Electron Lucent
- Black, dark areas
- Electron dense
24
Q
What is Scanning Electron Microscope (SEM)?
A
- Focused electron beam
* DOES NOT PASS THROUGH SPECIMEN - Image
* Scans only show the surface view
25
What is an Artifact?
1. Distortion of the tissue
* Ruins results
* Shrinkage, wrinkles, spaces, precipitates from the stain
26
What is Autoradiography?
1. X-ray to localize newly synthesized macromolecules
2. Specimen itself is the source of the radiation
27
What is Classical X-rays?
1. Specimen placed between source of radiation and the film
2. Absorption and scattering of radiation
28
What is Fibroblastic cells?
1. Bipolar/Multipolor
2. Elongated shape
3. Grow attached to a substrate
29
What is Epithelial-like cells?
1. Polygonal
2. Grow attached to substrate in discrete patches
30
What is Lymphoblast-like cells?
1. Spherical
2. Grown in suspension without attaching to a surface
31
What is Peroxidase?
1. Promotes oxidation of substrates
* Transfer of hydrogen ion to hydrogen peroxide
32
What is Phosphatase?
1. Removal of phosphate group from macromolecules
33
1. Kidney tubule treated histochemically
* Demonstrates Alkaline phosphastase
34
1. TEM image of Kidney cell
* Acid phosphatase
2. Dark areas
* Lead phosphate
