Histology- Exam #1 Flashcards
What is histology?
Science concerned with minute structure of cells tissues and organs in relation to their function.
What does fixation do?
Stops autolysis, prevents bacterial decompensation, stabilizes proteins
Is freezing used to fix samples?
Yes, but less frequently, freezing kills some cells and can leave artifact.
What kind of samples are usually fixed with freezing?
Nervous/ Adipose tissue
What occurs during processing?
Put in paraffin so it can be sliced thinly by microtome
What is staining based off of?
A variety of factors.
What kind of stains are there?
- electrostatic charges ( Acid or basic( most) Neutral stains: methylene blue picrate( rare), indifferent (Sudan III and Sudan IV ( is scarlet red))
- Supravital stains stain dead cells
- Vital stains stain live cells ( vital stains must be done quickly because if tissue dies tissue will not stain. (Methylene blue is vital stain)
What is the most routine stain?
Hematoxyline & Eosin are the most routine.
What is hematoxyline and what is its features?
Hemotoxyline is basic and attaches to negative charges ( Nucleic acids are neg charged) (stains purple) Natural dye that comes from Haematoxylon Campechianum ( found in mexico)
What are other basic stains?
- Toluidine blue ( mast cells)
- Methylene blue (vital stain)
- Fuscni Stains
What is eosin and some of its features?
Eosin is acidic and attaches to positive charges (Cytoplasmic structures are positively charged ( stains pink)
Acid stains what color in H+E?
PINK
Bases stain what color in H+E?
PURPLE
What are other acidic stains?
- Orange G
- Phyloxine
- Aniline blue
What are some artifacts that can be present in samples?
Crystal artifact in liver tissue, from freezing, autolysis- from lack of staining.
What should you base a size estimate off of for an object in a slide?
On size of a red blood cell. Which is dependent on animal.
What is the approximate sizes of RBC in the chicken, frog, dog, and goat?
‣ Chicken: 9.5 microns
‣ Frog: 10-24
‣ Dog: 5 microns.
‣ Goat: 2.5 microns.
What is this stain? What is it used for?
Trichome stain ( Massons, Mallory, ect, colors depend on tissue/ cell type, ( skin is a good one for this )
What is this stain? What is it used for ?
Wright-Giemsa and Diff Quick (Good Cellular Stain, Cytology/ blood smear)
What is this stain? What is a good use for it ?
Orecin: Good for seeing wall thickness in cells
What is this stain? What is a good use for it ?
Toluidine Blue Specific for mast cells ( stain because of high concentration of negative charges found in heparin in the granules)
What is this stain? What is it used for ?
◦ Prussian Blue/ Perls Stain: Shows iron deposits (Iron stains blue). (Good stain to study hemochromatosis in liver cells of captive Mynah birds)
What is this stain? What is it used for?
◦ Congo Red (polarizer): Will stain amalloid bright green, too much amalloid is not good for you.
What is this stain? What is it used for?
◦ Gram staining: Bacteria stains purple based on if it gram positive or negative.
What is this stain? What is it used for?
◦ Ziegler Neelsen( acid Fast)- Tb is acid fast
What is this stain? What is it used for?
◦ Silver Stains ( GMS, Gomori, ect): Stains fungal wall black.
What kind of stain is this? What is this used for?
◦ Periodic Acid Schiff Stain (PAS): Stains sugars ( fungus stains pink) Can also stain polysaccharides in tissue ( Goblet cells stain dark because they are rich in polysaccharides)
What is electron microscopy? What are some reasons it is not used as much?
- Higher resolution than light microscopy
- Requires special training, equipment and is expensive so not used as much.
- Fixatives are different then regular histology ( usually gold or other heavy metals) (Glutharaldehyde is most common)
What kinds of electron microscopy are there?
- Scanning EM: 3D structure, due to electron deflection.
- Transmission EM: 2D structure, goes through in one direction.
What is IHC?
Immuno Histo Chemistry
Target detection via Antibodies
In IHC, What are some important factors?
- Ab-Ag bond is specific, both are proteins.
- Immortal cells ( myeloma ( tumor of ab making cells)
What are Polyclonal antibodies? Monoclonal?
- Polyclonal ab: multiple epitomes (can attach to multiple places)
- Monoclonal Ab: single epitope (can attach to one place)
What are epitopes?
• Epitopes: part of protein that AB will attach to.
What is the difference between Direct IHC and Indirect IHC?
- Direct IHC: Primary AB is tagged
- Indirect: secondary ab ( targeting primary ab) is tagged ( secondary is flagged so you can see it)
What is the counter stain for IHC?
Hematoxylin
What is In situ hybridization?
- PCR: Amplification of DNA so you can detect/ see it. Can be done in real time. Specific to that sequence
- Usually more specific
- Ex: Mycoplasmosis is smallest self replicating prokaryote, and has sunny side egg colony
What is the process for cytologies? What types are there?
- Dry and stain, fixing is optional
- Various types: impression smears, fine needle aspirate, cytosine