Exam I Flashcards
Bichat’s types of membranes and their characteristics
Organic(vegetative)- life of heart and other organs and was regulated by collection of small independnt thoracic brains (ganglic NS)
Animal- composed of symmetrical organs and include habit and memory and ruled by wit and intellect
Marie Francois Xavier Bichat
- changed perception that organs contain tissues or membranes
- described 21 membranes w/o microscope
- divided membranes into organic and animal life
- brought down to tissue level
- disease attacked organs
Giovanni Battista Morgagni
- showed that diseases were due to organ lesions not humoral imbalances
- brought diagnosis down to anatomical level
Who won the Nobel Prize in Physiology/Medicine in 1906 and what did they do?
Golgi- created silver staining technique to stain nervous tissue.
Cajal- correctly interpreted the neural structure of the brain.
Definition of histology and histopathology.
study of the microscopic anatomy of cells and tissue of plants and animals.
histopathology- study of abnormal or diseased tissue
Humoral Pathology Concept and 4 humors
-concept that was prominent until the 18th century that suggested diseases were caused by an imbalance of the humors.
- blood
- phlegm
- yellow bile
- black bile
4 basic types of tissue
- epithelial
- muscular
- nervous
- connective
Rudolph Carl Virchow
- pioneered concept of pathology processes through application of cell theory
- “all cells from cells,” ex. maggots from trash
Refraction of light definition and equation
-the bending of light while traveling through another medium
refractive index= velocity of light/velocity of light inside medium
refractive index of air= 1
Light bending depends on:
- refractive index
- angle light hits surface
Refractive power
measure of how much a lens bends light waves. Measured in diopters. (1/focal length)
focal point
the point through which all parallel rays will pass after passing through each part of the lens
focal length
distance from center of lens to focal point
real image
-when an image is placed outside the focal point
- inverted
- can be projected onto a screen
- differs in size from real object (greatest magnification when short focal length and object is close to the focal point)
Virtual image
-seen when object is placed inside the focal point
- not inverted
- cannot be projected onto a screen
- can be magnified
resolution
the ability of microscope to distinguish between two objects as separate points, diameter of diffraction lines must be reduced
resolution equation
d=0.61(lambda)/n sin(a)
lambda= wavelength of light n= refractive index of medium (n air =1, n oil= 1.4) sin(a)= angle of cone of light entering aperture n sin(a)= numerical aperture
resolution in different microscopes
light microscope- highest is 0.5 micrometers (oil)
TEM- theoretically is 0.01A, but actual is 5-10A
Components of light microscope and what it does
light source, condenser (focuses light), stage(holds specimen), objective lens(first set lenses), ocular lens(second set of lenses)
Pros and cons of light microscope
- ability to magnify
- ability to resolve structural detail
- specimen must be thin
- little contrast in unstained specimen
Phase Contrast Microscope
- -phase shifts become visible
- can examine unstained cells and tissue
- can examine living cells
Fluorescent microscope
- detects molecules that emit light visible under UV
- detects naturally occurring fluorescent molecules such as vitamin A
- can detect stained molecules -> antigens/antibodies, fluorescent tracers in the body
Confocal Scanning Microscope
-increases resolution by eliminating out of focus light
-can recreate 3D images
-uses a laser and moves it across the specimen
-very thin images created (1micrometer thick)
-out of focus images are subtracted
-
Transmission Electron Microscope(TEM)
- utilizes a beam of electrons rather than light (heated tungsten filament)
- cathode
- anode(drives electrons through column)
- electromagnets
What are the series of steps used to prepare tissue for microscopic examination?
- fixing
- dehydration
- removal of alcohol
- embedding
What is fixing?
prevents further deterioration of the tissue and hardens the tissue prior to embedding and sectioning
What is one of the most widely utilized fixing agent and describe it according to lecture.
Formalin, can be used alone (buffer) or with other agents such as alcohol (causes shrinkage) and acetic acid (softens and counteracts shrinkage).
- reacts with amino acids to stabilize tissue
- not good if cytological detail is desired
What is an acid fixative good for?
An acid fixative fixes chromatin, nucleoli, spindle fibers, but not mitochondria or nucleoplasm.
What are the three acid fixatives discussed in class?
Carnoy’s fluid- useful for preserving glycogen in animal tissues
Zenker’s fluid- useful to view sharp detail, but must be washed carefully to prevent black precipitate.
Bouin’s fluid- widely used fixative, but must there must be prolonged and careful washing cycles
What is a basic fixative?
Good for mitochondrial staining, but it dissolves chromatin
What is the basic fixative discussed in class?
Zirkle-Erliki- requires long fixing time and washing under water
What are the two TEM fixatives?
glutaraldehyde- preserves proteins by cross linking them
osmium tetroxide- reacts with lipids and imparts electron density to cell and tissue structure
What is dehydration?
Removing all the water from the tissue before embedding and infiltrating with hydrophobic material. Consists of washing tissue in increasing strengths of alcohol. Ethanol dissolves neutral fats
What is clearing?
replacing the alcohol with an agent such as xylene, cedar oil, CCl4
What is embedding?
moves the tissue through three paraffin baths, placing in a mold, and then hardening in a cold water bath
Embedding for TEM
- use of monomeric resin
- resin is then polymerized
- less than 1mm3
What is sectioning? And how is this done?
Sectioning is slicing samples into smaller samples utilizing microtomes or sharp razors. For TEM, sections are cut into 50-150nm sections using a diamond blade. The sample is then placed on a copper mesh grid (electrons pass through grid)
What do you need to do to prepare a stain?
- Paraffin must be removed using xylene.
- xylene is removed using decreasing alcohol strength to water
- stains are applied and then dehydrated
- alcohol is removed with xylene
- drop of cement and cover slip applied
Hematoxylin and eosin
- behaves like basic dye due to mordant
- hemotoxylin stains dyes nuclear and cytoplasmic material blue
- eosin stains cytoplasm and extracellular blue
Examples of stains
- orcein and resorcin fuchsin stains used to reveal elastic material
- silver impreganation is useful for reticular fibers and basement membranes
- Sudans are good for lipids
Describe basic dyes and give examples.
- react with anionic groups such as phosphate, sulfate, and carboxylic groups (all three groups available at high pH)
- tissues that react with this basophilic
- methyl green, methylate blue, pyronine G, toluidine blue
Describe acid dyes.
- bind to cationic groups (amino acids) in tissue using electrostatic linkages
- all have slightly different properties and can be used in sequence to see different things
- aniline(collagen), acid fuchsin(cytoplasm), orange G(red blood cells)
Metachromasia
A change in dye color after reacting with tissue components
What do histochemical techniques do?
techniques used to study the chemistry of cells and tissues
Perls’ reaction
-demonstrate the presence of iron in tissue
Stains for lipids:
must use frozen sections because lipids are soluble in normal reagents. -Sudan IV, Sudan black, oil red O
Fuelgen reaction
detects aldehyde groups on deoxyribose using Schiff reagent
Periodic acid-Schiff reaction (PAS)
- PA cleaves carbon bonds to form aldehyde group
- Schiff reacts with aldehyde group turning pink
- detects polysaccahrides, glycoproteins, glycolipids, proteoglycans, glycosaminoglycans
- clinical application: biopsies of patients with glycogenoses (glycogen storage disease)
- best carmine does the same
RNA Stains:
-can be stained with basic dyes, but will need control slides to distinguish from other basophilic substances
Immunocytochemical stain definition and monoclonal antibodies
used to study specific antigens in tissue by using monoclonal (single immune response) antibodies.
-monoclonal antibodies are derived from activated B cell clones exposed to a specific antigen
Direct vs indirect labeling
direct: antibodies are conjugated directly with either fluorescent dye, gold or ferritin(TEM), or visible markers(light)
indirect: marker is attached to second antibody which is specific to the antigen that is being looked for
What tissue is found is most prevalent in the body?
epithelium
What are the different types of epithelium?
simple/stratified squamous, simple/stratified columnar, simple/stratified cuboidal, transitional, pseudostratified
What are the characteristics of epithelium?
- uniform geometric shape
- tightly bound together (tight junctions)
- little intercellular matrix
- display free surfaces (face lumen, surface, etc)
- cell exhibit polarity
- do not have blood vessels
- may be derived from the three germ layers
- can be innervated
- form secretory and excretory parts of glands
- line body cavities and cover body surfaces
- sits on top of basement lamina
What does cell polarity mean?
the cells have an apical end (top) that is in contact with the lumen and houses structures such as the microvilli, cilia, and sterocilia. The cells also have a basolateral side, which is where the cells come into contact with other cells. Connected via junction complexes and come into contact with basal lamina.
What are the components and functions of the basement membrane?
functions: selective filtration barrier, stabilization of tissue shapes, scaffold for embryogenesis and regeneration
components: composed of the reticular lamina and basal lamina. Basal lamina is next to the epithelia and is composed of type IV collagen and glycoproteins and is PAS+. Absent in lymphatic vessels and hepatic sinusoids. The reticular lamina is in contact with the connective tissue and is composed of argyrophilic fibers, reticular fibers, and glycoproteins.
What else can epithelia be identified by?
- keratin
- apical structures
Basolateral modifications
-maintain conc differences (tight vs leaky barrier)
+barrier type is determined by the the complexes that hold the cells together
-anchored to basal lamina
Where are simple squamous epithelia found?
- lines lumina of duct, vessels, and other tubular structures
- walls of alveoli, Bowman’s capsule, tympanic membrane, surface sof membranous labyrinth
Where are simple cuboidal epithelia found?
-kidney tubules, surface of ovary, pigmented epithelia of retina, kidney tubules, glands and ducts, terminal bronchioles, choroid plexus, anterior capsule of eye
Where are simple columnar epithelia found?
- ciliated: upper respiratory tract, uterine tubes, uterus, paranasal sinuses, central canal of spinal cord
- non-ciliated: cardia of the stomach, gall bladder, and excretory ducts of the glands
Where are stratified squamous cells found?
- heavily keratinized: epidermis and cornea
- lightly keratinized or non: esophagus, vagina, lining of mouth, tongue, and epiglottis
Where are stratified cuboidal epithelia found?
- rare
- anal mucosa, large excretory ducts, part of male urethra
Where are stratified columnar epithelia found?
- rare
- conjunctiva of eye, adult sweat glands, pharynx, epiglottis, parts of male reproductive/urinary tract
Where can pseudostratified epithlia be found?
- ciliated: trachea
- stereocilia: epididymis
Where can transitional epithelia be found?
urinary tract (urothelium)
What are the functions of epithelial tissue?
- covering and lining
- glandular
Simple, non-ciliated columnar epithelium is most likely to be found associated with which of the following?
digestive tract
Junctional complexes are associated with which cellular domains?
basolateral
Transitional epithelium is associated with which of the following systems?
urinary
Although not common, stratified cuboidal may be found in which of the following areas?
part of the male urethra
Lightly keratinized stratified squamous epithelium is characteristic of which of the following locations?
vaginal epithelium
What are the components of the basal lamina?
- secreted by the epithelial layer
- laminin
- fibronectin
- type IV collagen (does not form fibrils)
- entactin
- proteoglycans