BE 167L Lecture 5 General

BE 167L Lecture 5 General - BE 167L Bioengineering Lab...

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BE 167L – Bioengineering Lab Lecture 5
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Today Quiz 1 overview Why Do Cell Culture? Mammalian Cell Culture Cell Assay Techniques Upcoming Thursday: Three-dimensional Cell Culture and the Cellular Microenvironment Next Tuesday: Case Study 3
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Quiz 1 stats 70 points total Low – 16 (23%) High – 66 (94%) Mean – 45.6 (65%) Median – 45 (64%) SD – 11
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Problem areas Kohler Illumination Amino acid reactivity Importance of fluorescence spectrum overlap Designing scientific experiments Misconceptions about the use of stamps with cells
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What is Kohler illumination and how do you adjust it? A way to create uniform lighting on the sample If lighting is not uniform this should be one of the first things you check Some microscopes allow you to focus the condenser by moving it up and down until the light is defocused. The aperture should also be centered. Some microscopes have automatic kohler illumination fixed, so if it is off, you may have to call to have it fixed. Always try to read the manual of
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Focusing the condenser in brightfield Aligning the phase rings for phase contrast
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Carboxylic acids aren’t good nucleophiles You have to put a leaving group on it to activate it Good nucleuophiles will displace them COO- doesn’t displace NHS All ionizable amino acids don’t undergo the same
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Fluorescence is a continuous spectrum, make sure there are no critical overlaps Spectral overlap of the abs/ems can create unintended signals Depending on your filters and camera you may see more wavelengths than you think Emission from one and absorption by another can contribute incorrect signals λ / hc hv E = =
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How do you design a complete scientific experiment? (mini-projects this quarter) Define a hypothesis to be tested Outline experimental conditions that test it
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BE 167L Lecture 5 General - BE 167L Bioengineering Lab...

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