The BD FACSDiscoverTM S8 Cell Sorter is a new cytometer with a wide range of capabilities leading to special preferences recommendations.
The BD FACSDiscoverTM S8 Cell Sorter acquires traditional fluorescent parameters as well as a set of imaging parameters. There are a handful of recommendations we can offer to make the use of this data easier in FlowJo version 10.x. Preferences are accessed through the heart icon in the upper righthand corner of the workspace. The preferences for specific cytometers will be found under the Cytometers tab/button in the bottom row of options.
To set the preferences for the BD FACSDiscoverTM S8 Cell Sorter, find the name in your list list of cytometers and select it.
The BD FACSDiscoverTM S8 Cell Sorter fluorescent parameters will specify a Log transform function by default. We recommend checking the box for ‘Enable Transforms’ and setting a width basis of -100 to display the data on a biexponential scale, with a linear region of 100 units on either side of zero.
Next, we recommend using the parameter filtering options to filter out any parameter with -W, -H, or -T. These are each different methods of calculating a measure of fluorescent intensity from the measured electronic pulse. The -A parameter, which is the area of the pulse, is the most complete measurement, and -A is commonly the only measurement used for subsequent data analysis, making the other options redundant. Filtering them out pulse width (-W), height (-H), and time to peak (-T) can cut the displayed parameter list by 75%. However, alternate measures of some parameters can be useful for diagnostics and identifying single cells, so exempting the additional measures of side scatter seems useful and can be done by choosing ‘And not’ SSC.
*IMPORTANT NOTE: Changes to Cytometers Preferences will only affect data loaded into a new workspace.
Finally, the search tool in the parameter list becomes of paramount importance with these data. Typing in one of the image parameters (or alternatively one of the target antigen labels or specific parameter names), will limit the displayed parameters to those that partially match the search criteria. The example images below depict parameter lists after searching for either all of the light loss measurements (LightL), returning 16 imaging derived measurements, or searching for CD123, returning the 16 image derived measurements of CD123 and the one florescent measure.
