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Cell Bio Lecture 3

Cell Bio Midterm 1

What is cell fractionation used for? Lyses cells and separates organelles by size and density.
What is velocity sedimentation used for? Achieves a finer degree of separation and organizes cell components by size and shape.
What is equilibrium sedimentation used for? Achieves a finer degree of separation and organizes cell components by density.
What is chromatography used for? Proteins can be separated based on their properties (eg. charge, size, hydrophobicity, ability to bind small molecules).
Which type of chromatography is the method of choice for smaller molecules/proteins? HPLC
What is SDS-PAGE used for? Proteins migrate through a gel polyacrylamide matrix at a rate depending on charge, size, and shape; useful for membrane (hydrophobic) proteins; provides information about molecular weight and subunit composition.
What is mass spectrometry used for? Useful in identifying unknown proteins by mass/charge ratio; give an idea for the end protein amino acid sequence.
What is a yeast two-hybrid system used for? Useful in detecting protein-protein interactions using bait/prey resulting in transcription.
What is FRET/GFP used for? To see protein interactions/where in a cell a protein is located through labeling molecules with fluorochromes and seeing where fluorescence occurs.
What us X-Ray crystallography used for? It gives the 3D structure of proteins (provided that the protein can form crystals); usually used for larger proteins.
What is NMR used for? It gives the 3D structure of proteins without the need for a crystal; usually used for smaller molecules.
What is a Southern blot used for? Allows for the detection of a specific DNA sequence via probe hybridization; gives information about identity, size, and abundance.
What is PCR used for? Amplification of DNA; useful if the genome sequence is known.
What are expression vectors used for? Sequence of DNA is inserted into a vector (plasmid) and cloned for virtually unlimited amounts of protein expression.
What is RNA interference (RNAi) used for? Useful in figuring out what a gene does via knockout and degradation of specific mRNAs and viewing the consequences in a living organism.
What are microarrays used for? Allow for simultaneous analysis of thousands of RNAs through use of probes which bond to mRNA and monitor expression; probe sequences must be known in advance. Can compare genes under different environmental conditions.
Bacteria and mitochondria are generally the smallest objects which can be seen with a _____ microscope. light
_____ light microscopy allows for the visualization of density/cell structure. phase-contrast
_____ light microscopy takes advantage of light scattering as it passes through a specimen. dark-field
Define selective staining. Sections can be stained with dyes which have affinity for specific cellular components - hematoxylin has affinity for - charge molecules, eosin has affinity for + charge molecules.
What is in situ hybridization used for? Visualization of the distribution of RNAs in a sample of tissue with fluorescently labeled probes.
Describe fluorescence microscopy. Molecules are labeled with fluorescent markers which absorb one wavelength of light and emit another. It is used to detect specific proteins/molecules in the cell.
Define fluorescent nanoparticles. Fluorochromes which are more stable and fade less quickly than traditional methods; ie. quantum dots.
How are antibodies utilized in microscopy? Antibodies are labeled with fluorescent dyes; the antibody combines with an antigen and allows for location of molecules in the cell.
Describe confocal microscopy. Pin-point illumination of a single point in a specimen. Eg. seeing where are virus goes.
T or F: Electron microscopes give lower resolution than light microscopes. False
Describe electron microscopy. Specimens are stained and put through an electron beam/vacuum; gives fine cellular structure.
T or F: Immunogold microscopy is analogous to antibody detection with fluorescent microscopy. True
What is one pro and one con to immunogold microscopy? Allows to locate multiple proteins in one sample, but can only detect antigens at the surface of the section.
T or F: Scanning electron microscopy can only produce 3D structure on the surface of the specimen. True
What is scanning electron microscopy typically used for? Visualization of whole cells/tissues.
Describe electron microscope tomography. Allows for the specimen to be tilted and viewed from different angles in order to perceive depth; 3D images are combined and taken at the average.
What are the advantages of GFP? GFP can be produced by the cell. Cells are dynamic so it is possible to visualize movement of molecules in the cell; moniters gene expression.
What are fluorescent indicators used for? Allow for the measurement of concentration of a substance in the cell (eg. Ca) at different locations by binding to the molecule of interest.
What are five different methods of getting substances into cells? Micropipette injection, electroporation, lipid fusion, DEAE dextran, biolistics
Created by: goberoi