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4 Steps To Validate Flow Cytometry Antibodies And Improve Reproducibility

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Until we have access to well-validated recombinant antibodies produced under tightly regulated conditions, researchers need to exercise good judgment regarding these critical biological reagents. These 4 steps will help ensure that your results are consistent and reproducible. This will both reassure your reviewers that your data is of high quality, and allow for researchers at other institutions to successfully replicate your results. In addition, identifying antibody duds early on will save you time and money in the long run. Don’t shirk the work of ensuring your antibodies are working correctly and targeting the right proteins.

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How To Use Flow Cytometry To Measure Apoptosis, Necrosis, and Autophagy

Flow Cytometry measuring apoptosis, necrosis, and autophagy
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Using flow cytometry and a host of different reagents, it is possible to tease out how your cells may have died. Using these tools, you can readily eliminate the various suspects and come to your conclusion as to how your treatment may have killed your cells of interest. Here are some reagents to consider when measuring apoptosis, necrosis, and autophagy.

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Flow Cytometry Protocols To Prevent Sample Clumping

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Good flow cytometry depends on a high quality, single cell suspension. If the cells put through the instrument are not of high quality, the ensuing data will be difficult to analyze. Likewise, if the sample is clumpy, one will not be able to readily distinguish cells of interest from the clumps they are attached to. Sample preparation becomes the critical first step in any flow cytometry experiment. To get high quality results, follow these 3 sample preparation steps.

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How To Compensate A 4-Color Flow Cytometry Experiment Correctly

4-Color Flow Cytometry Experiment | Expert Cytometry | multicolor flow cytometry
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Compensation in flow cytometry is a critical step to ensure accurate interpretation of data. It is also one of the areas that’s steeped in mystery, myths and misinformation. Manually adjusting the compensation values based on how the populations look, or so-called ‘Cowboy Compensation’, is not the correct way to determine proper compensation. The best practices for compensation involve following some very specific rules. Here are 4 steps to correctly compensating 4+ color flow cytometry experiments.

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What Is A Statistical Analysis T-Test And How To Perform One Using Flow Cytometry Data

t-test statistical analysis of flow cytometry data | Expert Cytometry | t-test formula for data analysis
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The T-Test compares the differences between the means of two populations to determine if the null hypothesis should be rejected. At a minimum, to perform the T-Test, one needs the means and standard deviations of both populations, and the number of measurements. The researcher also needs to set the threshold value, also termed the α. Then, you will compare this threshold to the P-value. If the P-value is greater than the α, there is no significance in the data. However, if the P-value is less than the α, there is significance in the data. Here’s how to run a T-Test.

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