A type of light microscopy with a max resolu5on of
0.2 um
Useful for…
Viewing 5ssues that absorb light
Not useful for…
Example of image
Transparent or colorless 5ssues that don’t absorb light Set up
Condensor and objec5ve Sample is stained
DIC/
Nomarski
Align the light microscope for color illumina5on and then place the DIC elements in the op5cal path, when lights don’t interfere they appear dark, while when there is an edge of the structure, the beams are altered and when the beams combine via the second prism either construc5vely to produce a bright spot or destruc5vely to produce a dark spot. Don’t require staining or fixing
Phase
Contrast
Transparent/
Colorless
specimens to enhance contrast
Once the objec5ve phase for non light and the annulus are absorbing concentric and overlapped specimens, we can view the enhancing the specimen, Don’t require contrast from staining or fixing shades of gray to shades of black and white, living samples are observed Specimens are surrounded by a halo which obscures fine structures this halo is an ar5fact caused by illumina5on of the annulus, Shows the loca5on of specific molecules in the cell by tagging the molecules of interest with fluorescent molecules such as stains or an5bodies. max resolu5on of 10 nm
Slicing destroys material, light emiTed by sample comes from molecules above and below the plane of focus so the image appears blurred Conven(onal
Fluorescence
Microscopy
Producing high resolu5on pictures of fine structures by enchaining contrast at interphases. Op5cal sec5ons are produced.
Shows the exact loca5on for a point of interest in the cell
Tissue samples since they are thick/ Highly pigmented cells
No5ce the sharp edges on the sample, the bright side and the shadowed side but this can be easily resolved by rota5ng the sample to the other side
No5ce the halo around each sample, this is a feature of phase contrast
No5ce the different fluorescence colors used to stain different parts, the image is blurred due to the limita5on of focusing
BrighOiled microscope with the addi5on of
Polarizer is inserted between light source and condenser, spliQng prism beam in the condenser tart, DIC combining prism at the back of the objec5ve and an analyzer in the infinity space in the tube lens.
BrighOiled microscope with the addi5on of special phase objec5ve containing a ring, and a condenser annulus instead of a diaphragm, annulus located in the condenser tarot.
An5bodies or Stains that are specific to certain material is used. Then brighOield microscopy is done.
Fluorescnet
proteins are some5mes expressed in the cell. Descrip(on
