CTools012411lecture08

CTools012411lecture08 - Biology 172 Lecture 8: Monday,...

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Lecture 8: Monday, January 24, 2011 Biology 172 Today’s Outline Announcements •  This Week in Discussion:  Enzymes; no quiz • Exam 1 is Mon., Jan. 31,   6-8 pm  (150 pts) How Cells are Studied Cell Structure - Prokaryotic vs. Eukaryotic Cells - Cell Size - Structures in All Cells Properties of Eukaryotes - Information Processing - Matter Processing - Synthesis - Export - Recycling
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How Cells are Studied Microscopy Cell fractionation Mutational analysis Selective disruption or inhibition Specific labeling of cells/parts
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Light Microscopy Bright Field (Transmitted light) Fluorescence
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Conventional fluorescence image
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Confocal fluorescence image
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Transmission Electron Microscopy Scanning Electron Microscopy Nucleus Most bacteria 10 m 1 m 0.1 m 1 cm 1 mm 100 µm 10 µm 1 µm 100 nm 10 nm 1 nm 0.1 nm Human height Length of some nerve and muscle cells Chicken egg Frog egg Most plant and animal cells Mitochondrion Smallest bacteria Viruses Ribosomes Proteins Lipids Small molecules Atoms Unaided eye Light microscope Electron microscope nucleus Most bacteria
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Electron Microscopy Cilia from a rabbit trachea 1 µm Scanning EM Transmission EM Uses a beam of electrons instead of light to create an image.
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DIFFERENTIAL CENTRIFUGATION Low-speed centrifugation Medium-speed centrifugation Supernatant Pellet High-speed centrifugation 1. Start with uniform cell homogenate in centrifuge tube. 2. After low-speed spin, pellet contains large components (including nuclei and cellular debris ). Transfer supernatant to new tube. 3. After medium-speed spin, pellet contains medium components . ( mitochondria., peroxi- somes, chloroplasts, ) Transfer supernatant to new tube. 4. After high-speed spin, pellet contains small components ( internal membranes, ribosomes ).
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DENSITY GRADIENT CENTRIFUGATION Lower-density solution Higher-density solution Sample 1. Add sample to tube of variable- density solution. 2.
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CTools012411lecture08 - Biology 172 Lecture 8: Monday,...

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