The FACS Discover S8 spectral imaging sorter is a new cytometer with a wide range of capabilities leading to special preference recommendations.
The S8 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 cytometers are under the Cytometer tab.
To set the preferences for the FACS Discover S8, find it in your preference list and select it.
The S8’s fluorescent parameters are recorded as log currently. We recommend checking ‘on the ‘Enable Transforms’ and setting a width basis of -100 to display the data on a biexponential scale and 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 in most cases enough, 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.
Finally, the search tool in the parameter list becomes of paramount importance with this data. Typing in one of the image parameters (or alternatively one of the targets), particularly if you have filtered, gets you a very manageable list to work with. In these examples we search for either all of the light loss measures, returning 16 imaging derived measurements, or for CD123 returning the 16 image derived measurements of CD123 and the one florescent measure.
