I’d like to (re)introduce Flow Core users to the Amnis ImageStream MK II, an imaging flow cytometer. With the ImageStream, individual cells are illuminated by both a brightfield source and 405 nm, 488 nm, 561 nm, and 642 nm lasers at 20x, 40x, or 60x magnification. The ImageStream combines the throughput and quantitative ability of flow cytometry with the detailed imagery [...]
I am not sure how widely appreciated it is that the temperature at which you stain your cells can make a huge difference with some markers. This was shown years ago in this publication, among others. Below is a recent experiment we did testing the staining of CCR7 in human peripheral blood. Conclusion, staining on ice completely destroyed any CCR7 staining. [...]
Most of you folks probably don't pay enough attention to this sort of thing. But the Cell Sorters are approved for BSL2+ sorting. However this ideally involves a significant level of PPE which is often times not utilized. In order to accommodate a wider array of sample types and materials, while at the same time keeping sorter operators (both staff and [...]
Presentation -There are now 3 Beckman Coulter Cytoflex instruments on campus. These instruments have different detectors, electronics, software than what most people are used to, i.e., BD and FacsDiva. -Flowjo default settings are generally set up to analyze data collected on BD instruments. -Getting data from the Cytoflex instruments to be visualized properly may take some adjustments to settings you may [...]
There are a few factors that get evaluated when designing immunophenotyping panels. Antigen density Brightness of fluorophore Fluorophore overlap (Compensation) Expression of protein (eg. is antigen A also expressed on the same cells as antigen B) It takes a lot of experience to be able to put these all together. Over the last little while Tessa has been comparing a bunch of [...]
I've been getting a lot of questions from folks about what dyes can and can't be used together in the UV and Violet detectors. I've put together a little presentation that will hopefully shed some light on a few potential options that will make your life easier a little easier by incorporating the UV laser into your panels. As always, let [...]
I am in the process of developing a training seminar specifically tailored for compensation. I came across this presentation and thought I would post it since it's pretty darn good. It's relatively complicated so if doesn't make sense right away that's ok. We will go over this material when the seminar is finished Take home messages: Voltages don't make a difference [...]
Counting Statistics and the Poisson distribution In the process of flow cytometry cells (events) are presented for analysis at random and their distribution will be described by Poisson statistics. Any sub-set of cells will also be distributed at random within the parent population and we can consider the statistics of these separately. The essential feature of Poisson distributions is that if [...]
The intended audience for most of these "rules" is the universal flow cytometry core facility staff member. They were developed for training and educational purposes over the last 10 years. However, in order for everyone's expectations to be met within the University of Utah core, I believe this list is applicable for both core facility staff as well as the client. [...]
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