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Instrument Quality Control For Reproducible Flow Cytometry Experiments

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The flow cytometer is an integral component of any flow cytometry experiment, and special attention should be paid to ensuring that it is working correctly and consistently. As an end-user, the researcher should be able to sit down at a machine and know that it is performing the same way today as it was yesterday and last week. Equally important is that if any changes in instrument performance have occured, the end-user knows how they have been addressed and corrected, rather than letting them fester and potentially affect the results. Quality control measurements can include a variety of targets, such as PMT sensitivity, laser alignment, fluidic stability, background issues, and more.

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Experimental Controls For Reproducible Flow Cytometry Measurements

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With the increased focus on reproducibility of scientific data, it is important to look at how data is interpreted. To assist in data interpretation, the scientific method requires that controls are built into the experimental workflow. These controls are essential to minimize the effects of variables in the experiment so that changes caused by the independent variable can be properly elucidated. Getting into the mindset to improve the reproducibility of flow cytometry experiments requires a hard look at the appropriate controls to use in each experiment.

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Statistical Challenges Of Rare Event Measurements In Flow Cytometry

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It is necessary to sort through hundreds of thousands or millions of cells to find the few events of interest. With such low event numbers, we move away from the comfortable domain of the Gaussian distribution and move into the realm of Poisson statistics. There are 3 points to consider to build confidence in the data that the events being counted are truly events of interest and not random events that just happen to fall into the gates of interest.

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Procedural Limitations That Impact The Quality Of Rare Event Flow Cytometry

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Stem cells, circulating tumor cells, and minimal residual disease in cancer patients were all discovered through the power of rare event flow cytometry. When preparing for rare event analysis, sample preparation and data analysis must be taken into account at the beginning. How will we stain our cells? How will we analyze our cells? What controls will we use to help us identify our rare events? What statistical methods do we use to analyze our results? Here are 5 procedural limitations that impact the quality of rare event flow cytometry data and how to optimize your assay to get the best results possible.

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How to Optimize Flow Cytometry Hardware For Rare Event Analysis

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Preparing for rare event analysis requires an understanding of the power and limitation of the instrument to be used. From how fast to run the fluidics, to how the signal is processed to the number of gates that can be used in the sorting experiment, each factor impacts the outcome of the experiment.

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