Exam 1 w1-4

interactions of light w/ tissue components

brightfield, phase contrast, normarski/DIC, fluorescence, confocal

interactions of electrons with tissue components

transmission, scanning

ability to perceive the presence of an object

ability to distinguish between two objects that are close together

the smallest distance at which two objects can be distinguished

the image size relative to the object size

magnification which yields no further useful information or resolution

• fixation (crossling proteins), 2. embedding (supporting medium), 3. sectioning, 4. staining

H = blue = binds to DNA and RNA, E = red = binds to proteins

a detectable marker covalently attached to an antibody that binds to a specific protien/antigen

the primary antibody recognizes a specific protein/antigen and a secondary antibody covalently attached to a detectable marker reveals the primary antibody

secondary antibodies are coupled to enzymes to extremely amplify the signal for rare proteins

fluorescent molecules are used to stain specific molecules or organelles

secondary antibodies couples to fluorophores

a protein sequence that can be expressed and will fluoresce in any organism; sequence is inserted downstream of the promotor of the gene of interest and the resulting protein is expressed with a GFP attached to it

visualize proximity of interacting proteins based on the light emission, green light = interaction between proteins

provides quantitative kinetic data to measure protein diffusion, transport, and dissociation rates; an area of the membrane is bleached and the diffusion of protein recovers the area –> rate of recovery=rate of protein diffusion

laser scanning of a focal plane deep within a sample

confocal images slices are taken in z-stacks that can be flattened into a 2D image or assembled as a 3D reconstruction

e- pass through a specimen to form a 2D image

e- scattered or emitted by a specimen form a 3D image of the sample surface features

in a dish