Microscope Flashcards
What is Chromogenic Staining
Stains nuclear DNA purple (haematoxylin) and Cytoplasmic Proteins pink (Eosin)
What is DIC
Differential interference contract. Is a light microscopy with 2 light beams, is related to refractive index. Shows major organelles. For thick and to observe small details
What is Phase contrast
Refractive and un-refracted light, dark outline. for Thin cells
What to use on moving molecules
Use a time-scale
Steps for dead specimens
First fix the cell, embed in wax, slice and view under microscope
Types of light microscopy
Phase-Contrast and Nomarski and bright field
What is FACs
fluorescence- activated cell sorter
Flow cytometry
fluorescence activated cell sorter isolates specific cell types, can use FACs to seperate specific cells
Example of FACs sorting
Sorting T cells: T cells fluoresce both green and Red as as a tagged antibody specific for certain proteins is added they have THY 1.2 ad CD3
Differential Centrifugation
Based on mass, spins and heaviest goes to bottom
What is immunofluorescence
detects proteins using fluorescence tagged antibodies direct and indirect.
Confocal Microscopy
Does one plane therefore allows to not have blurred images
fluorescence microscopy
Can view specific molecules in live cells e.g. using stains like eosin an hematoxylin
When same mass what can you use
density gradient
What can you do if you can’t get a specific antibody
You can epitope tag the c or n terminus
What can epitope tagging an organelle show
Show if it is in the cytoplasm or inside an organelle
How to tell the orientation
C or N terminus and have a tag which will fluoresce when attaches and use a detergent digiton or Triton-X, can see if in organelle or just in cytoplasm
What is TEM
Transmission electron microscope
What does TEM do
Detects differences in density, high resolution. Focus on high electron densities rather then florescence. Electrons emitted from a filament and accelerated in an electric field. Then condenser lens focuses on, then scanning and objective specimen an dthen detector
What is SEM
Scanning electron microscope
What does SEM do
metal-coated speciemen, in a high vaccum, allows veiwing of surface features
What do they do to cells and tissues for EM
Cut into thin sections
What is Immunoelectron Microscopy
Use an antibody with a heavy metal tag to veiw proteins
Viewing shape with EM
Can stain sample with heavy metal to view shape
