Term
| Adv/Disadv. of organ perfusion, explant cultures, tissue slices, minces, and homogenates |
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Definition
- Adv: closer to in vivo situation
- Disadv: specific cell's contribution lost as cell response averaged out
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Term
| Laser capture microdissection |
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Definition
- isolate specific cells or components from tissue
- laser dissection and catapult under microscope
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Term
| procedure for single cell isolation |
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Definition
- Digest with trypsin and collagenase
- Filter
- Purify
- Purity determination by microscopy and/or cell markers
- if long term use, then culture
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Term
| Primary cell culture (adv/disadv) |
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Definition
- adv: provides info about cell's contribution
- tissue availability and shorter lifespan of cells due to telomerase depletion and shortening of telomeres
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Term
| Immortalized cells (what, how, adv, disadv) |
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Definition
- Tumor cells
- immortalize normal cells by introducing the gene encoding telomerase catalytic subunit or oncogene that inactivates the cell cycle checkpoint mechanisms
- disadvantage: lose features of primary cells over time
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Term
| Fluorescence activated cell sorting (FACS) |
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Definition
- separate diff. cell types by tagging them with fluorescent molecules
- cells labeled, into droplets, excited with laser, fluorescence measured, droplets given + or - charge based on measurement, diverted to collection tube by charged plates
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Term
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Definition
- cytoplast = cell minus nucleus
- nucleoplast = nucleus alone
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Term
| production of hybrid cells (why, how) |
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Definition
- to immortalize (Ab. production)
- fuse differentiated normal cell with mouse tumor cell using PEG or inactivated viruses, fused cell called heterocaryon, selective medium allows only heterocaryons to survive, enter mitosis, nuc envelopes dissassemble, chromosomes mix, immortal hybridoma cell line
- Fig 8-7 pg 509 in the cell
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Term
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Definition
- break open cells in isotonic buffer
- differential centrifugation to obtain crude fractions
- purification by continuous or discontinuous sucrose density gradient centrifugation
- determine purity by microscopy and/or marker enzymes
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Term
| cell fractionation by centrifugation |
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Definition
- cell homogenate> low speed> pellet: whole cells, nuclei, cytoskeletons > sup. med speed > pellet: mitochondria, lysosomes, peroxisomes > sup. high speed > pellet: microsomes, small vessicles > sup. very high speed > pellet: ribosomes, viruses, large macromolecules
- Fig 8-10 pg 511 in "the cell"
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Term
| mag and resolution of LM and TEM |
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Definition
- LM: mag - 1000X; res - 0.2 μm
- TEM: mag - 250,000X; res - 2nm
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Term
| 4 types of LM used to study living cells |
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Definition
- bright field
- dark field
- nomarski differential interference contrast
- phase contrast
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