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Final notes
 

Final notes

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    Final notes Final notes Document Transcript

    • Final Notes:<br />
      • Cellular Chemistry
      • Types of sugars
      • Sucrose – Aglucose + Afrucrose = alpha glycosidic
      • Lactose – Bglucose + Bgalactose = beta glycosidic
      • Maltose – Aglucose + Aglucose = alpha glycosidic
      • Types of bonds
      • DNA/nucleotides – phosphodiester
      • Maltose/disaccharide – glycosidic bond
      • Enzyme/amino acid – peptide bonds
      • Metabolism
      • Catabolic – breakdown – release heat
      • Anabolic – creation of molecules
      • Ana- means “to go back to”
      • Energy
      • G = H -TS
      • Studying Cells
      • Cell-lines
      • Some cells reach confluency – a single layer of cells, stopped growing due to social signals of the cells that there will not be enough resources if the cells keep dividing
      • Primary culture – initial cells placed on slide
      • Secondary culture – ‘immortal’ cells produced from primary cultures after confluency. (some cells die after confluency)
      • Oncogenic transformation – cells not as dependant on substrate and divide on top of one another
      • Terms used in microscopy
      • Magnification – lets you see how large you can view an object while keeping the components as distinct objects. How big can the object be while maintaining objects as distinct?
      • Resolution
      • R= (0.61*lamba)/(n * sin(angular aperture))
      • a factor that measures how close two points can be in a sample and still be distinct.
      • R=194 nm. This means that two objects that are 200nm apart can be seen as distinct objects. If they are closer than this, they will not be seen as distinct structures.
      • Different types of light microscopy
      • Cell often appears transparent when there isn’t manipulation
      • Brightfield stained
      • Contrast for organelles – must be dead
      • Brightfield unstained
      • Fluorescent
      • Stains used but can be alive
      • Tagged proteins for induced expression
      • DIC – differential interference contrast imaging
      • Insertion of nomarski optic - A prism that splits light into two separate waves through specimen. Changes wavelength of light and the interference leads to increased contrast, largest difference is the edges of cells and organelles.
      • Phase contrast
      • Differentiation of organelles and differences between them w/out fixing
      • Good way to see edges of cell
      • Vesicle transport in cells
      • Takes advantage of physics of light and the characteristics of it as a waveform. When two waves are in phase, they are at the same frequ and combine to create a better resolution. If out of phase, reduced intensity of light. When in phase, the unstained cell will disrupt one of the light waves creating a slowed speed for the wave form.
      • Confocal Imaging
      • Thicker samples and image optical layers for 3d images.
      • TEM vs SEM
      • 2d vs 3d
      • Fluorescence Microscopy
      • Lots of dyes to look at parts of cells at the same times.
      • Only limited by number of filters one can place in microscope.
      • Source of light is ultraviolet passed through an excitation filter, which removes wavelengths of light not useful in expressing the fluorescence.
      • Dyes are chemically derived that bind carbohydrates, lipids, DNA.
      • One doesn’t need to use antibodies if the dyes bind directly.
      • Proteins most often identified by fluorescently linked antibodies because few dyes directly link to proteins.
      • Primarily, the fluorescent microscope does not have a condenser like in a light microscope. Must be thin samples.
      • confocal
      • has allowed us to use laser light to image specific layers/regions within a thick sample.
      • Images individual parts and put together to rebuild the picture.
      • Membranes
      • ∆Gc = RTln[in/out] + zFVm
      • F = 23,062
      • Vm = membrane potential in volts
      • R = 1.987
      • Types of ATPases
      • P type (for phsphorylation)
      • Hydrolyze ATP
      • Na+/K+
      • V-type (vacuole)
      • Used to pump H+ across membrane for pH maintenance
      • F-type (factor)
      • ATP synthases
      • ABC-type