Histology Flashcards
What are the four basic tissues and function of each one?
1) Epithelia: cover and lines surfaces. I.e what lines intestines.
2) Connective tissue: packing, support, connecting.
3) Muscle: contractility.
4) Nerve: Irritability, conduction
Preparation steps of tissue for microscopy.
1) Fixation
2) Dehydration in alcohol.
3) Embedding
4)Trimming/sectioning
5)Removal of paraffin
6)Rehydration
7)Staining
8) Mounting sections on glass slide
9)Viewing the tissue (light microscope)
What preparation steps for microscopy affect the appearance of tissue?
1)Fixation: sample placed in a chemical solution that preserve the tissue by cross-linking proteins and inactivating degradative enzymes.
2) Dehydration in alcohol: removes water
3) Embedding: tissue placed in a small mold with melted paraffin (wax) and allowed to harden.
4)trimming/sectioning : paraffin block is trimmed to expose the tissue for sectioning (slicing) on microtome
7) staining: colors the cells that is completely colorless. Acid and basic dyes used to form salt linkages with molecules in tissues
Acid dye (most common )
Eosin. Binds with basic part of the cell
Stain blue!
Basic molecules bind with acidic dyes and stain red.
Basic dye (most common )
Hematoxylin. Binds with the acidic part of the cell
Stain red!
Acidic molecules bind with basic dyes and turn blue.
Basophilic
(Acidic molecules )Cell components such as nucleic acid with a negative charge (anionic), stain with basic dyes.
Acidophilic
(Basic molecules)Cell components that contain lots of basic molecules, such as protein with many ionized amino group, stain with acidic dyes.
What color is the cytoplasm stained? why?
Cytoplasm would be pink, bc it has lots of proteins which are basic molecules. Basic molecules love acidic dyes.
What color is the nucleus stained ?
Blue. It contains nucleic acid (acidic) and that loves basic dyes.
Luxol blue and hematoxylin
Luxol blue is a better dye to visualize nerve tissue
Trichrome
Visualize connective tissue better. Stains Ct in green and blue.
Why don’t fat and mucus stain well with H and E?
Bc they get washed out in preparation process.
Hydrophobic structures also tend to remain clear; these are usually rich in fats, e.g.adipocytes,myelinaround neuronaxons, andGolgi apparatusmembranes.
H&E staining dissolves fat out of the cell
Periodic Acid Schiff Reaction (PAS)
Best for staining mucus, microvilli, basement membrane and glycogen granules.
Sudan Black
Best for fat and myelin
What are the artifacts caused by preparation ?
Artificial space, wrinkles in tissues and precipitate of stains.
What causes artificial spaces in tissue ?
Shrinkage due to fixation, dehydration and embedding.
Loss of molecules( glycogen and lipids) that were not retained after fixation or removed during dehydration.
Electron Microscopy
Transmission electron microscope (TEM): shows inside of cell.
Scanning electron microscope (SEM): shows surface of cell.
Light grey: electron Lucent
Dark grey: electron dense.
Nucleus Structure
Membrane-limited.
Nucleus envelope: outer and inner membrane. outer membrane faces the cytoplasm, house ribosomes and is continuous with RER at certain sites.
The inner membrane faces the genetic material and is supported by nuclear lamina on its inner surface.
Nuclear pores on the surface of nucleus to aid with transport.
nucleolus in the center.
Nucleus functions
Cellular regulation: houses genetic material which direct cellular activities.
Production of ribosomal subunits that are exported into cytoplasm for ribosome assembly.
Function of nuclear lamina
Support of inner nuclear membrane
nucleus stability.
chromosome attach.
Mutation in nuclear lamina
Muscle dystrophy ( genetic defect condition)