Term
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Definition
| break up tissue sample with proteases; machine seperates them by charge |
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Term
| what is a transformed cell? |
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Definition
| one that divides indefinitely |
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Term
| what are the three requirements for cell culturation? |
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Definition
1. growth media (salts, glucose, any amino acids they can't make for themselves, etc.)
2. serum/growth factors
3. (most animal) physical support structures |
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Term
| light microscopy vs. fluorescent microscopy vs. electron microscopy |
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Definition
light: far away/larger image
fluorescent: inside the cell
electron: close up/more detail |
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Term
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Definition
| the ability to define objects seperated by small distances |
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Term
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Definition
| numerical aparture / the light gathering power of the lens. together with wavelength of light, defines resolution |
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Term
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Definition
| NA (light-gathering power) determined by angular aperture and refractive index |
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Term
| direct protein labelling vs. indirect protein labelling |
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Definition
direct: attaching GFP to antibodies to show you where a protein is
indirect: attaching GFP to one antibody that transfers it to the antibodies already attached to a protein |
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Term
| FACS again (what they do, not how they work) |
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Definition
seperate different cells physically, then plot on a chart - every dot is a different cell
(also it stand for Flow-Activated Cell Sorting) |
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Term
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Definition
| computer analysis of a cell illuminated by fluorescence microscopy that takes multiple pictures from small points to assemble a whole, clear image without it being fuzzy/out of focus from having too big of an area |
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Term
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Definition
| a form of deconvolution where a small point of light bounces back through a pinhole; in this way, only light from the plane you're looking at is admitted and it keeps the image in focus |
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Term
| what are the weaknesses of confocal microscopy as a method of deconvolution? |
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Definition
1. photobleaching (color leeches out after focus by a camera)
2. phototoxicity (kills the sample) |
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Term
| multiphoton fluorescence microscopy |
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Definition
| dye requires two photons to be excited; this is very unlikely except on the point of focus |
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Term
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Definition
| fluorescence recovery after photobleaching - photobleach something, then watch the tagged proteins come back to study the rate of movement |
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Term
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Definition
fluorescence resonance energy
two proteins are tied to two fluorescent dyes, one of which excites the other; excite one to see if the other lights up (if so, they're linked) |
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Term
| why is electron microscopy able to achieve greater resolution than light microscopy? |
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Definition
| electrons have a very short wavelength |
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Term
| transmission electron microscopy |
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Definition
| cells are fixed and stained with heavy metals; beam of electrons is passed through; scatter pattern is marked on screen |
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Term
| what is metal shadowing and what is it usually used for? |
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Definition
| a form of electron microscopy that is usually used for examining the details of a cellular surface; you spray with a thin layer of metal at an angle so you can make out the details via shadow |
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Term
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Definition
| 3d image by computer combining various 2d images from different angles |
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Term
| scanning electron microscopy |
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Definition
| passing a beam of electrons over, but NOT through, the cell - you can see the outside surface, cell interactions, etc. |
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Term
| what are two factors by which things are split up in differential centrifugation? |
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Definition
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Term
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Definition
| breaking the cell up into small parts (necessary step before differential centrifugation) |
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Term
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Definition
| the study of how genes interact with each other |
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Term
| 3 different types of differential centrifugation |
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Definition
1. basic differential centrifugation - larger, more dense things come out at lower speeds, and smaller, less dense ones at higher speeds
2. velocity centrifugation - separated out by the speed at which they move through a fine sucrose gradient
3. equilibrium centrifugation - spun until they reach buoyancy with surrounding sucrose - seperates purely by density |
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Term
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Definition
| all the proteins that are expressed in a given cell |
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Term
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Definition
| i.d. all the proteins expressed in a cell; interactions; etc. |
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Term
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Definition
| separates proteins out based by size/charge; then they're cleaved by a protease, analyzed in a mass spectrometer, and compared to a database of known size/mass ratio |
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Term
| shot gun approach to proteomics |
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Definition
| skips the gel electrophoresis part - goes straight to digesting the proteins with protease, then sequences everything at once by tandem mass spectronomy |
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