To conclude our series on rare event analysis, it is time to discuss the statistics behind rare event analysis. The first 2 parts of this series covered the hardware aspects of measuring rare events and some specific recommendations for gating/analysis of rare events.
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 ...
Having dealt with the hardware issues related to rare event analysis in the first part of this series, it is time to turn to our second focus: how samples are prepared.
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 ...
â€śNot everything that can be counted counts and not everything that counts can be counted.â€ť â€” William Bruce Cameron (but often misattributed to Albert Einstein)
What does this quote mean in terms of flow cytometry? Flow cytometry can yield multi-parametric data on millions of cells, which makes it an excellent tool for the detection of rare biological events â€” cells with a frequency of less than 1 in 1,000.
With the development and commercialization of tools such as the Symphony, the ZE5, and others which can measure 20 or more fluorescent parameters at the same time, researchers now have the ability to ...
For those new to flow cytometry, compensation is confusing at best and terrifying at worst. Likewise, those who have been doing flow cytometry since the analog ages may be holding on to practices that, while suited to the analog instruments, should be left to the annals of history. As such, a lot of time is spent discussing compensation and the best practices for this critical process.
There are 3 rules that guide proper compensation, and theyâ€™ve been written about extensively since they first appeared in the â€śDaily Dongleâ€ť in 2011. It is always good to review and, importantly, there are some ...
Congratulations, your grant has been funded! Now comes the hard part â€” performing the work that you are being funded to do. This means generating data and publishing papers. What was that hypothesis again? It must be in the grant somewhere, right?
For the sake of this blog, the grant is to study the effects of Cordilla Virus, which is known to cause lipid membrane flipping on CD8+ T-cells. This flipping results in phosphatidylserine expression on the outer membrane, causing infected cells to be phagocytosed by macrophages. A lead compound, Masiform D, has been identified that shows promise in reducing the ...
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