Difference between revisions of "Team:MIT/results"

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<p>Graphs, explanation of results</p>
 
<p>Graphs, explanation of results</p>
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<h2> Interpreting the Results of Cytometry</h2>
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<p>We used CytoFlow, a software developed by our iGEM advisor, Brian Teague, to analyze the results of our HEK transfections.
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Import samples from file generated by the flow cytometer.</p>
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<p> 1. Assign each sample a unique set of conditions, which could be Booleans, Categories or Numbers. These distinctions can be taken from your transfection planning document. Name carefully, as these variable will be what you use to process your data later
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<p>Gate the live cells based off their forward scatter height (FSC-H) and side scatter width (SSC-W). These are the laser scatters caused by the cells passing through the cytometer. You want to select a population of cells that have a high FSC-H and SSC-W. [Example figure goes here]</p>
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<p> Set your transfection threshold, by measuring the amount of fluorescence outputted by your transfection marker. Transfection efficiencies typically range from approximately 10-85%.  [Example figure goes here] </p>
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<p>Bin your cells by the amount of plasmid in each cell. Binning is a method of standardizing your output across different cell plasmid concentrations. If a cell receives more plasmid, we would expect it to have a higher fluorescence output across all colors. We divided our population into 40 transfection bins. [Example figure goes here] </p>
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<p>Based on your variable and output fluorescence, you want to group your cells by variables and compare the fluorescence in every case.
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</p>

Revision as of 01:00, 29 October 2017

Results

Graphs, explanation of results

Interpreting the Results of Cytometry

We used CytoFlow, a software developed by our iGEM advisor, Brian Teague, to analyze the results of our HEK transfections. Import samples from file generated by the flow cytometer.

1. Assign each sample a unique set of conditions, which could be Booleans, Categories or Numbers. These distinctions can be taken from your transfection planning document. Name carefully, as these variable will be what you use to process your data later

Gate the live cells based off their forward scatter height (FSC-H) and side scatter width (SSC-W). These are the laser scatters caused by the cells passing through the cytometer. You want to select a population of cells that have a high FSC-H and SSC-W. [Example figure goes here]

Set your transfection threshold, by measuring the amount of fluorescence outputted by your transfection marker. Transfection efficiencies typically range from approximately 10-85%. [Example figure goes here]

Bin your cells by the amount of plasmid in each cell. Binning is a method of standardizing your output across different cell plasmid concentrations. If a cell receives more plasmid, we would expect it to have a higher fluorescence output across all colors. We divided our population into 40 transfection bins. [Example figure goes here]

Based on your variable and output fluorescence, you want to group your cells by variables and compare the fluorescence in every case.