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Histology Outline 1
Histology Class Chapter 1
Term | Definition |
---|---|
Histology | Study of tissues (and cells). It is a microscopic study |
Tissues | A group of cell performing similar functions. |
Extracellular matrix | Molecules which combine to form larger structures. Made of fibers. |
How slides are made for light microscopy Step 1 | Tissue sampling - post mortum dissections, biopsy samples, surgical excisions. |
How slides are made for light microscopy Step 2 | Fixation - Immediately after tissue sampling. It prevents enzymatic degradation of tissues by autolysis killing any pathogens such as bacteria, viruses, and fungi. It also preserves cellular structure. |
How slides are made for light microscopy Step 3 | Embedding & sectioning -Dehydration before imbedding. |
How slides are made for light microscopy Step 4 | Staining - Dyes are acidic or basic. |
Acidic dye examples | Eosin, Orange G, Acid fuchsin |
Acidic dyes are ________ charged and they stain a _____ color because it binds to _______ which are ______ | Net Negatively. Orange G*. Cations. Amino groups of proteins |
Basic dyes examples | Hematoxylin |
Basic dyes are ________ charged and they stain a _____ color because it binds to _______ which are ______ | Net positively. Toludine blue*. Anions. Phosphate groups, nucleic acids, sulfate groups, glycosaminoglycans, and carboxyl groups of proteins. |
The glycosaminoglycans are part of the ______ | Extracellular matrix |
Acidic dyes bind to cationic groups in a reaction called ______ | eosinophilia |
Basic dyes bind to anionic groups in a reaction called ______ | Basophilia |
Primary stain H&E | acidic and basic |
Stain Cartilage | green/blue color |
Mallory stain | |
Periodic Acid Schiff (PAS) | Stain polyssacharides |
Silver stain | Stains particular fibers blue-purple-black color |
oil red O, Osmic acid and Sudan black | Oils and lipids |
Light microscope condenser | focuses light |
Light microscope objective lenses | magnify objects |
Light microscope eyepiece | 10x magnitication |
Resolution | ability to distinguish between two objects. |
Phase contrast microscopes also use light, but vies unstained specimens | Light passes through different structures and different speeds- some structures are darker than others |
Differential interference microscopes | light microscopes in 3D |
Other types of light microscopes | Polarizing, confocal, fluorescence |
Polarizing microscopes | Light microscopes with polarized filter and a second filter- layer upon layer |
Confocal | focuses light even more in small areas |
Fluorescence | Uses fluorescent dies which are kidney dyes |
Electron microscopy-TEM & SEM advantage | increased resolution to about 0.2-0.5 nanometers |
EM | 5000x-120000x magnification |
Electron dense areas in EM | darker |
Electron lucent areas in EM | lighter |
Principle of EM | electromagnetic fields can deflect a beam of electrons, after passing through a specimen. |
Images are always _______ in EM | Black and white |
SEM | Has no penetration. Scans specimen in 3D magnifies 10000x That way you can see cells and organs. |
Freeze fracture | Tissue is rapidly frozen the hit with an object such as a razor blade and then viewed. This is used to see the internal structures |
Structures are sectioned in planes | Longitudinal Transverse/cross section Oblique |
Longitudinal | Parallel to longest section |
Transverse/cross section | perpendicular to longitudinal |
Oblique | Between long and cross section |
Serial section | Serves to ascertain true structures and figure out what the specimen looks like |
Tangential or grazing section | .... |
Is what appears as a single layer of cells, truly just one layer? | Probably not |
Is an element of human error involved in making slides? | Of course |
Faulty techniques | artifacts or flaws |
Degeneration of tissues | distorted images - not fixed right after sampled |
Shrinkage of tissues | Empty spaces - could appear because of the heat of wax, fixative, or/and alcohol |
Thick staining | precipitate of staining granules |
Mishandled tissues | Folds or wrinkles |
Two most commonly used chemicals | Gluteraldehyde and 4% Formaldehyde (or formalin = 17% aqueous solution of formaldehyde) |
link amino groups of proteins and since proteins are important it will link 3D structure of cell/tissue and they will be maintained. | .... |