Term
| Requirements for growing cells in culture |
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Definition
| surface, incubators, antibiotics, media, vitamins |
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| components of the media for growning cells? |
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Definition
| 9 essential amino acids. Plus three other amino acids that are only synthesized by specialized cells in intact animals. Also needs glutamine (nitrogen source) |
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Definition
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| what do animal cells need to grow? |
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| cells use to bind to adjacent cells and to components of the extracellular matrix such as collagen of fibronectin |
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| established by normal animal tissues or whole embryos |
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| lineage of cells originating from one initial primary culture |
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| made up of very rare cells that undergo oncogenic mutations that allow them to survive and continue dividing until their progeny overgrow the culture. They will grow indefinitely if it is appropriately diluted and fed with nutrients, these cells are immortal |
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Definition
| refers to the rare cells in a population of primary cells may undergo spontaneous oncogenic mutations, these cells are able to grow indefinitely |
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| Cells in immortalized lines |
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Definition
| have chromosomes with abnormal DNA sequences.They also usually have more chromosomes then the cell from which they arose from = aneuploid |
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| How are CHO cells unique? |
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Definition
| CHO lines and its derivative have fewer chromosomes than their hamster progenitors |
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Definition
| is used to separate cells which cannot easily be separated by equilibrium density centrifugation. Florescent dye linked to an antibody for the specific cell marker tags the cells that are to be separated. Flow cytometer quantifies the number of cells that are fluorescent |
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Definition
| fluorescence-activated cell sorter- analyze and sort cells. Droplets have to contain one cell each. Cell surface markers are specific to one type of cell, the cell mixture is incubated with a fluorescent dye linked to an antibody to the specific cell surface molecule, and the |
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Term
| significance of growing cells in three dimensions |
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Definition
| Three dimensional growths provide a support infiltrated with components of the extracellular matrix. The apical aspect of the epithelial sheet lines the lumen, the basal side of each cell is in contact with the extracellular matrix |
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Definition
| are produced via introduction of an antigen. The antigen causes B cells to produce antibodies to that epitope. Each epitope specific B cell forms a clone which produces the same antibody. |
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Definition
| grow rapidly and are immortal, they produce monoclonal antibody encoded by its B-lymphocyte parent. They are made from the fusion of a myeloma cell with a normal antibody-producing cell |
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Definition
| employed in affinity chromatography to isolate and purify proteins from a complex mixture |
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Term
| what limits the resolutions of the light microscope |
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Definition
| Limit of resolution of a light microscope using visible light is 0.2 um. Light microscopes can never resolve objects that are less than 0.2um apart or reveal details smaller than 0.2um. |
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Term
| the purpose of fixing, sectioning and staining in microscopy |
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Definition
Fixing helps in light/electron microscopy to reveal fine details of structure, the solution contains chemicals that cross-link most proteins and nucleic acids
Sectioning is when the tissue sample is embedded in parafilm and cut into sections
Staining is used to visualize the main structural features of cell or tissue, the stain binds to molecules that have specific features |
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Definition
H = hematoxylin binds to basic amino acids on many different kinds of proteins
E = eosin binds to acidic molecules
H & E visualizes different parts of the cell |
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Term
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Definition
| sensitive to Ca2+, binds Ca2+ in the cell, binding is proportional to cytosolic Ca2+, can be used to quantify rapid changes in the fraction of fura-2 that has a bound Ca2+ion and thus in the concentration of cytosolic Ca2 |
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Definition
sensitive to the H+ concentration, monitors the cytosolic pH of living cells
Fluorescence microscopy is a type of light microscopy, it is the most versatile and powerful technique for localizing molecules within a cell by light microscopy |
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Term
| how fluorescent dyes can be used to visualize the levels of specific molecules in living cells |
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Definition
| Modern techniques are configured to pass the excitation light through the objective into the sample and then selectively observe the emitted fluorescent light coming back through the objective from the sample |
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Term
| Immunofluorescense microscopy |
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Definition
| detects specific protein with an antibody to which a fluorescent dye has been covalently attached. Antibody is covalently attached to fluorochrome; complex binds to corresponding antigen then light up when illuminated by exciting wavelength. |
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Definition
| is when two proteins can be visualized simultaneously, using antibodies made in different animals and second antibodies labeled with different fluorochromes. Used to study two proteins at one time |
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Definition
| uses computational methods to remove fluorescence contributed from out-of-focus parts of the sample. Images display impressive detail without blurring |
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Definition
| is green fluorescent protein comes from jellyfish; it contains serine, tyrosine, and glycine sequence whose side chains spontaneously cyclizes to form green-fluorescing chromophore when illuminated with blue light. |
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Term
| How is GFP used to study cells using fluorescence microscopy |
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Definition
| It can be used to study cells by tagging proteins for GFP and covalently linked to GFP as part of the same polypeptide. Used to visualize the protein of interest and assess dynamics of various cell treatments |
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Term
| the two types of confocal microscopy |
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Definition
| (1)Laser-scanning confocal microscope and (2) spinning disk confocal microscope |
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| What does confocal microscopy allow for? |
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Definition
| Confocal microscopy provides an accurate three dimensional representation can be computationally generated |
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Definition
| total internal reflection fluorescence microscopy- used in a complex mixture of fluorescent structures to see those structures that are close to the thickness of the cover slip |
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Definition
| fluorescence recovery after photobleaching- used to show dynamics of the cellular components |
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Definition
| Forster resonance energy transfer- utilized two fluorescent proteins in which the emission wavelength of the first is the same as the excitation wavelength of the second |
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Term
| Two types of electron microscopy |
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Definition
| TEM (transmission electron microscopy) and SEM (scanning electron microscope) |
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Definition
| can localize specific proteins via antibodies associated with heavy metal marker |
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Definition
| reveals the surface features of specimens |
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Definition
| controls movement of molecules in and out of the cell and functions in cell-cell signaling and cell adhesion, located around the outside of the cell |
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Definition
| surrounded by a double membrane, generate ATP by oxidation of glucose and fatty acids, located in the cytoplasm |
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Definition
| have an acidic lumen, degrade material internalized by the cell and worn-out cellular membranes and organelles, located in animal cells only in cytoplasm |
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Definition
| a double membrane, encloses the contents of the nucleus; the outer membrane is continuous with the rough ER, located outside the nucleus |
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Definition
| nuclear subcompartment where most of the cell’s rRNA is synthesized, located inside the nucleus |
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Definition
| synthesizes lipids and detoxifies certain hydrophobic compounds, located outside of nucleus |
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Definition
| functions in the synthesis, processing, and sorting of secreted proteins, lysosomal proteins, and certain membrane compounds, located outside the smooth ER |
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Definition
| processes and sorts secreted proteins and fuse with the plasma membrane to release their contents, located outside the rough ER |
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Definition
| store secreted proteins and fuse with the plasma membrane to release their contents, located near Golgi complex |
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Definition
| detoxifies various molecules and also breaks down fatty acids to produce acetyl groups for biosynthesis, located outside the Golgi |
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Definition
| form networks and bundles that support cellular membranes help organize organelles, and participate in cellular movement, located near the cell wall/outside of cell |
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Definition
| increase surface area for absorption of nutrients from surrounding medium, located on the outside of animal cells |
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Definition
| composed of largely cellulose, helps maintain the cell’s shape and provides protection against mechanical stress, located on plant cells |
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Definition
| stores water, ions, and nutrients, degrades macromolecules and functions in cell elongation during growth, located in plant cells |
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Definition
| carry out photosynthesis, are surrounded by a double membrane and contain a network of internal membrane bound sacs, found in plant cells |
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Term
| Differential Centrifugation |
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Definition
| filters cell homogenate at increasingly higher speeds, supernatant is poured off and centrifuged at a higher speed. |
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Term
| Equilibrium density gradient centrifugation |
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Definition
| separates cellular components according to their density, after the fraction is resuspended; it is layered on top of a solution that contains a gradient of a dense non-ionic substance. Equilibrium density gradient centrifugation is used to further separate the pellets of differential centrifugation |
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Term
| how drugs are used to study cellular process in cell biology research |
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Definition
| Inhibit specific and essential processes of cells |
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Term
| How siRNA can be used to study cellular processes |
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Definition
| Treatment with siRNA leads to destruction of target mRNAs and hence “knock-down” of the encoded protein |
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Term
| mechanism by which siRNA functions |
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Definition
| If the single stranded siRNA sequence can base pair exactly with a target mRNA sequence, the argonaute protein-RNA complex cleaves the target mRNA, which is then degraded |
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Term
| Does siRNA have a natural cell function? |
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Definition
| Originally used in defense |
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Term
| What does the Smooth ER do? |
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Definition
| synthesizes the phospholipids that form the plasma membrane |
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Term
| the pH of lysosomes is lower than that of the cytosol because of the action of |
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Definition
| H+ and Cl- transport proteins in the lysosomal membrane |
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Term
| Oxidation of fatty acid occurs in |
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Definition
| mitochrondria and perioxisome |
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Term
| Which organelle is composed of series of compartments that function to sequentially modify proteins and lipids |
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Definition
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Term
| The phenomenon in which a chemical absorbs light at one wavelength and emit light at a different wavelength is |
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Definition
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Term
| what is used to visualize subcellular organelles in living cells? |
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Definition
| differential intereface light microscopy |
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Term
| osmium tetroxide is commanly used to |
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Definition
| stain specimens for tranmission electron microscopy |
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Term
| TO visualize cells by immunoflorescense microscopy cells must be |
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Definition
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Term
| fluorsence properites of SNARF 1 can provide information on the |
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Definition
| concentration of H+ ions in specific regions of the cell |
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Term
| The first pellet is made of what (in differential centrifugation) |
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Definition
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Term
| a method of disrupting the cell using sound |
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Definition
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| Ultracentrifugation isolated on the basis of |
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Definition
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Term
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Definition
| an immortal immune cell that cannot produce antibodies |
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Term
| Factors that are necessary for growth of animal cells in culture provided by serum? |
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Definition
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Term
| What are produced by animal cells in culture and other cells to adhere to culture disk |
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Definition
glycoproteins, collagen, hyalauronic acid
NOT phospholipase A |
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Term
| characteristics of a tranformed cell (3) |
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Definition
anueploidy, ability to differentiate, and presence of integrated viral genes
NOT tight junctions |
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Term
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Definition
| can knockdown (eliminate) the expression of specific genes, can leads to degradation of target mRNA and leads to increased gene expression of targeted genes |
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Term
| What is used to study protein-protein interactions in vivo |
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Definition
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Term
| Which technique combines protein specific anitbodies and fluorsence to detect proteins in fixed cells? |
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Definition
| immunoflorescense microscopy |
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