Term
1) What is the difference between direct immunofluorescence and indirect immunofluorescence?
2) How are these techniques different from using GFP? |
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Definition
1) Direct immunofluorescence involves incubation of tissue with antigen-specific antibodies coupled to flourophores such as FITC or Alexa dye. Indirect immunofluorescence involves a fluorophore-coupled secondary antibody, which targets a primary antibody that is bound to a target antigen. This method requires less primary antibody, and is therefore often preferred.
2) Using GFP is somewhat similar to direct immunoflourescence in that it involves fusing GFP to a target protein and visualizing that protein through the fluorescence of the GFP. |
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Term
| Why are hematoxyylin & eosin stains used in light microscopy? |
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Definition
Hematoxylin is a positively charged, basic stain that interacts with negatively charged acidic components of the cell such as heterochromatin, nucleoi and rough ER (appear blue to purple)
Eosin is a negatively charged, acidic stain that interacts with positively charged, basic structures such as cytoplasm and ECM components (pink) |
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Term
1) When might you use electron microscopy (EM) instead of light microscopy?
2) What are the two types of EM and how are they different? |
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Definition
When you need a resolution of over 1000X (can reach 100,000X)
2) Transmission Electron Microscopy allows one to study of the inner structure and contours of objects (tissues, cells, virusses)
Scanning Electron Microscopy is applied to visualize the surface of tissues, macromolecular aggregates and materials (individual organelles) |
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Term
| What are the primary functions of the cell nucleus? |
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Definition
1) contain chromatin
2) regulate import/export of factors that modulate transcription
3) modulate protein/RNA synthesis by controlling transcript production |
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Term
| What are the differential functions of the rough and smooth ERs? |
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Definition
Rough ER- synthesis of integral membrane and soluble proteins destine for secretion or for delivery to biosynthetic organelles.
Smooth ER- lipid synthesis and carbohydrate and lipid metabolism. Also acts as a reservoir for intracellular calcium that can be released by second messengers. |
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Term
| What is the primary role of the Golgi Aparatus? |
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Definition
the Golgi Apparatus is held near the nucleus via microtubule attachments.
It is important for Post-translational modifications such as proteolytic processing, maturation of oligosacharides, lipid attachment to proteins and phosphorylation. |
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Term
| What is the normal travel route of a protein from the inside to the outside of the cell? |
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Definition
| ER- ERGIC- cis Golgi- medial Golgi- trans Golgi- TGN- PM |
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Term
| The plasma membrane can have a number of types of projections, in addition to membrane receptors. What are these and what are they composed of (hint. there are 3 types)? |
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Definition
1) Microvilli- actin-based finger projections
2) Cilia- microtubule-based, long and motile structures
3) Stereocilia- actin-based, very long similar to cilia in appearance and found in inner ear and epididymis |
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Term
| What are the primary functions of mitochondria? |
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Definition
| Synthesize ATP and NADH for cell using energy derived from electron transport and OXPHOS |
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Term
| What processes are peroxisomes particularly import for? |
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Definition
1) breakdown of fatty acids and detoxification of various molecules (alcohol and bile acids) producing hydrogen peroxide as a byproduct.
Remember, peroxisomes are also membrane-bounded organelles |
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Term
| What are the primary functions of the cytoskeleton. What are the 3 cytoskeletal filament systems? |
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Definition
Cytoskeleton is important for mechanical support, chromosome movements, cell locomotion, vesicular transport and phagocytosis.
1) Actin- composed of two strands of f-actin in an alpha helix (4-7nm diameter)
2) Microtubule (14 columns of alpha and beta tubulin forming a hollow tube)
3) Intermediate Filament- lamins (lamina), vimenten (fibroblast), desmin (muscle), glial fibrillary acid protein (astrocytes and schwann cells), peripherin (neurons), cytokeratin (epithelial) |
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