From Purity To Biosafety, Understanding The Cell Sorting Process
Cell sorting is a combination of a numbers game (Recovery), quality of output (Purity) and speed. For any experiment, the end goal is going to be measured by these three characteristics, and as soon as one of these measures is more heavily favored, the other two must be compromised in some manner.
When designing a sorting experiment, start with the question of what will the cells be used for after sorting, and how many cells will you need for those experiments? That will set the minimum recovery that is needed. The second question is how pure do you need the cells? The requirements of the downstream assay will also dictate the purity needed.
The cell type being used will, in part, dictate the speed of sorting. Smaller cells can be sorted faster because a smaller nozzle can be used.
When you start a cell sort it’s important that you are aware of the downstream analysis and assays that you want to run. This will determine how you perform the sort and how you determine if your sort was successful or not.
Successful cell sorting involves balancing recovery, yield and speed. What do these three terms mean and what influences each of these factors?
Best Flow Cytometry Cell Sorting Practices
As a researcher, you want to achieve the best cell sorting possible. So, how can you achieve that? There are clear strategies you can use to achieve great cell sorting results, including finding your ideal sample concentration, using magnetic sorting to enrich your population, suspending cells in the right buffer to avoid cell clumps, changing your instrument settings when sorting small cells, and optimizing your sample preparation and instrument when sorting large cells.Read More
5 Essential Beads For Flow Cytometry Experiments
Flow cytometry is designed to measure physical and biochemical characteristics of cells and cell-like particles using fluorescence. Fundamentally, any single-particle suspension (within a defined size range) can pass through the flow cytometer. Beads, for better or worse, are a sine qua non for the flow cytometrist. From quality control,to standardization, to compensation, there is a bead for every job. They are important — critical, even — for flow cytometry.Read More
Use This Preparation Guide For Accurate Flow Cytometry Results
There is a lot of preparatory work that must be done before the first flow cytometry experiment can be attempted. Each step builds upon the previous one and extends where the assay is going. Be prepared for some trial and error in this process, and don’t expect perfect results the first time around. An educated user is a good user, and makes the SRL staff’s job that much easier. The partnership between investigator and SRL staff is a rewarding one, when both parties work together to achieve the ultimate goal of generating excellent data and sort results that help answer the biological question being tested.Read More
5 Essential Calculations For Accurate Flow Cytometry Results
Flow cytometry is a numbers game. There are percentages of a population, fluorescence intensity measurements, sample averages, data normalization, and more. Many of these common calculations are useful, but surrounded by misconceptions. This primer will help you decide which calculation to use, when to use it, and how to interpret the results.Read More