detailed 3D structures of sub-cellular macromolecular objects no requirement for ultra-thin serial section reconstruction an example is CAT
Scanning Electron Microscopy (SEM)
type of electron microscope that images a sample by scanning it with a high-energy beam of electrons in a raster scan pattern. (outside of a cell) The electrons interact with the atoms that make up the sample producing signals that contain information about the sample's surface topography, composition, and other properties such as electrical conductivity.
What is the most widely used procedure to separate fractions?
Centrifugation will separate sub-cellular structure and macromolecules into fractions by shape, size and density.
What are the different ways that cells and tissues may be broken?
high-frequency sound, mild detergent, forcing through a small hole, shearing cells with a rotating plunger
broken cells and tissue in one solution
repeated centrifugation at higher speeds will fractionate the cell homogenate.
also known as isopycnic density gradient centrifugation, is a type of centrifugation procedure widely used in biochemistry to separate molecules based on their isopycnic point (their buoyant density).
What is SDS-Page? What does it do?
SDS polyacrylamide gel electrophoresis proteins are separated in the electrophoresis by size (not shape, or charge)
(protein immuno-blot) detects specific proteins in tissue homogenate gel electrophoresis to separate proteins by 3-D structure or denatured proteins by the length protein transferred to a membrane and detected using antibodies specific to the target protein
detects RNA (isolated mRNA) used by molecular biology research to study gene expression
detects specific DNA sequences developed by Edwin Southern
separates mixtures of proteins, nucleic acids, and other molecules the principal that molecules dissolved in a solution will interact with a solid surface ion-exchange chromatography, and gel filtration chromatography, affinity chromatography
a flow cytometer can ID cells by measuring the light they scatter; sorts cells of a particular type from a mixture
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