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2. basic methods in histology
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- 1histology is a the anatomical study of the microscopic structure of animal and plant tissues.LM can cover most like human egg, most plant and animal cells. nucleus, most bacteria and mitochondria
EM covers smallest bacteria, viruses, ribosomes, proteins, lipids - 23 important variables in microscopy are 1. magnification 2. optical resolution 3. contrast what are they?1. magnification: amount fhb image of an object is increased
2. optical resolution: clarity, the separation distance at which 2 objects in an image can be apart and still distinguished as separate
3. contrast: to see something against its background; use bright field/fluorescent - 3LM is ______ in magnification? resolution down to? includes 3 types which are bright field, fluorescent, and phase-contrast what are its basic parts?LM is ______ in magnification? x1000
resolution down to? 200 nm
basic parts? eye piece, objective lens, condenser lens, light source, arm - 4in EM, what is magnification? resolution down to ? black and white image only transmission EM uses _____ and scanning EM uses?in EM, what is magnification? 1 million x
resolution down to 0.2 nm
transmission EM uses electron beam
scanning EM uses
scattering electrons on the sample - 5fixation what are some common fixatives 1. 2. 3. (which ones in LM and EM)?1. alcohol
2. formaldehyde (10% formalin) in LM
3. gultaraldehyde in EM - 6what does fixatives do ?fixatives arrests biological activity, prevent autolytic degradation and preserve (putrefaction), and render the cells more amenable to staining
- 7histology helps visualize tissue structures and understand function, LM and EM are complementary what are 2 or 3 traits of LM?polychromatic (several colours) stains over a wide area
easy sample preparation and operation - 8EM, what are 2-3 traits (good) about it? dis (3)?superior resolution and magnification,
see sub-cellular features
over much smaller areas
disadvantages: less flexible, complex sample preparation, costly - 9LM - brightfield microscopy what is a problem? and its solution (3)? new problems (3)problem: most cells are colourless and transparent
solution: stain with dyes to visualize structures, therefore fix cells/tissue, embed in wax to support tissue
new problems: processing alters cell structure and create artefacts, only snapshot of dead cells, only 2D of a 3D structure - 10fixation and embedding of tissues, thin sections for visualization, what are the hard medium used in LM and EM? what does fixing tissue require prior to embedding?in LM, use paraffin wax
in EM, plastic resin
-fixed tissues require dehydration with organic solvents prior to embedding - 11what processes causes artefacts (distortions in cell/tissue architecture, eg. shrinkage)?fixation, dehydration, emdedding, and secitoning, and staining all cause artefacts
- 12what machine is used for tissue sectioning?microtome
movement of its arm, create specimen embedded in wax or resin - 13staining techniques, haematoxylin and eosin, what types of dyes is each and what do they stain?haematoxylin, a basic dye stains acidic structures a purplish blue. eg. nuclei, ribosomes and rough ER (high DNA/RNA content)
eosin, acidic dye, stains basic structures red/pink eg. cytoplasmic proteins
animal cell -> nuclei stains blue, cytoplasm pink - 14what is giemsa? what does it stain?standard method for staining blood cells,
nuclei stain dark blue to violet
and cytoplasm pale blue - 15what is toluidine blue?basic stain that stains acidic components various shades of blue
- 16what is mason's trichrome?stain nuclei/basophilic structures blue
collagen stains green or blue
and cytoplasm stains bright red
eg. connective tissue technique - 17what is periodic acid-schiff reactions (PAS):stains complex carbohydrates
eg. mucin (glycosalated proteins) in goblet cells (column-shaped cell in respiratory and intestinal tracts, secretes the main components of mucus) - 18what other staining techniques? 1. 2. 3.1. immunohistochemistry (look at specific proteins, use antibodies to label)
2. detection if a cell status
3. labelling of a specific protein - 19look at cell structures and morphology to understand function. for ex. simple columnar epithelium functions? stratified epithelium functions?simple columnar epithelium functions? absorptive, secretory surfaces
stratified epithelium functions?
protective function, eg. skin - 20the prostate gland. prostate is ~ a size of a walnut in young adults, what is it responsible for? what does it consist of? important site of pathology in men of what age?responsible for: storage and secretion of seminal fluid, and muscle contraction during ejaculation
consists of branched tubuloacinar glands embedded in a fibromuscular stroma
>50 years old - 21prostate has transition zone, central zone, and peripheral zone, each surrounds?prostate has transition zone surrounds urethra (5%), central zone surrounds ejaculatory ducts (20%)
, and peripheral zone surrounds bulk of glandular tissue (70%) - 22the prostate gland, what is corpora amylacea? a feature of?what is corpora amylase
- lamellated glycoprotein masses
a feature of increasing age - 23the prostate gland, between secretory cells and basement membrane lies? its hard to detect on route H and E, it expresses what protein? therefore uses what to detect?lies a layer of flattened basal cells
expresses a keratin, uses an antibody to detect, and immunoperoxidase reaction to highlight this layer - 24what would a cancerous prostate cell include?loss regular structures
dysplasia (abnormal cell growth)
and hyperplasia (increase in cell #) - 25breast (glandular) adenocarinoma would include loss regular structures, dysplasia and hyperplasia too. what is the term used to describe the process when a timor (neoplasm) spread to surrounding tissue?invasion
- 26what is the term used to describe the process of a neoplasm invading lymphatic and/or blood vessels?metastasis
- 27what is dysplaic? what is angiogenesis?dysplasia: enlargement of an organ or tissue by the proliferation of cells of an abnormal cell type. as a development of disorder or early stage of cancer
-angiogenesis: development of new blood vessels - 28what authority regulates activities (removal, storage, use and disposal of human tissue) and make sure consent?human tissue author
tissue removed and stored for diagnosis does not fall under the act
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- biol Biol 133 Fichas sobre 2. basic methods in histology, creado por Jenny Lee7018 el 02/06/2016.
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- Jenny Lee7018
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