BIS104 Slide3 - Lecture 3 1. Microscopes and cell studies...

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Lecture 3 1. Microscopes and cell studies -continued 2. Lipids and Membranes
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Making gene replacements in mice genome Altered genome Altered DNA Figure 8-65 Molecular Biology of the Cell (© Garland Science 2008) Embryonic stem cells
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Figure 8-65 Molecular Biology of the Cell (© Garland Science 2008)
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Figure 8-6 Molecular Biology of the Cell (© Garland Science 2008) More about ES cells Personalized ES cells for tissue repair Somatic cells
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CELL STUDY - MICROSCOPES
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Figure 9-1 Molecular Biology of the Cell (© Garland Science 2008)
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Light microscope Can magnify up to 1000 X Bacteria ~1 µ m in diameter Animal cells ~10 µ m in diameter Maximum resolution: ~ 0.2 µ m Figure 9-3 Molecular Biology of the Cell (© Garland Science 2008)
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Two ways to obtain contrast in light microscopy Stains absorb certain wavelength -color images can be made Figure 9-7 Molecular Biology of the Cell (© Garland Science 2008)
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Some cells can be visualized by chemical staining Urine collecting duct of the kidney Figure 9-11 Molecular Biology of the Cell (© Garland Science 2008) Section of plant root
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Two ways to obtain contrast in light microscopy Stains absorb certain wavelength -color images can be made Phase changes can be made visible using phase-contrast or Normaski microscopy Figure 9-7 Molecular Biology of the Cell (© Garland Science 2008)
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Bright field Dark field (+Fluorescent stains) Phase-contrast Normaski (differential interference contrast) Four types of microscopy Figure 9-8 Molecular Biology of the Cell (© Garland Science 2008)
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Fluorescence microscope Figure 9-13 Molecular Biology of the Cell (© Garland Science 2008)
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Figure 9-18 Molecular Biology of the Cell (© Garland Science 2008) Labeling proteins with Ab and fluorescence markers Indirect Immunocytochemistry
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The same area stained with fluorescent Ab against tubulin (using indirect immunocytochemistry) Figure 9-17 Molecular Biology of the Cell (© Garland Science 2008) transmission electron micrograph of the peripheral of an epithelial cell -showing the distribution of microtubules and other filaments.
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Green: spindle microtubules Red: centromeres Blue: DNA Cell in mitosis Figure 9-15 Molecular Biology of the Cell (© Garland Science 2008)
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How to visualize thick samples? -Make very thin slices
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Drosophila embryo: Conventional vs Confocal fluorescence microscopy Figure 9-21 Molecular Biology of the Cell (© Garland Science 2008)
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Pollen grains-3D reconstruction of confocal images Figure 9-22 Molecular Biology of the Cell (© Garland Science 2008)
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Transmission electron microscope Figure 9-42 (part 1 of 2) Molecular Biology of the Cell (© Garland Science 2008)
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Most biological materials are still transparent…. .
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Figure 9-46 Molecular Biology of the Cell (© Garland Science 2008) Localizing protein using EM- immunogold EM Gold particles -electron dense, appear as back dots under EM G
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Scanning electron microscope -Images of the surfaces Figure 9-49 (part 1 of 2) Molecular Biology of the Cell (© Garland Science 2008)
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BIS104 Slide3 - Lecture 3 1. Microscopes and cell studies...

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