Identifying channels available in combination with Fura Red calcium dye.

<p>Expanded human T cells were loaded with 1 μM Fura Red, AM for 30 minutes. Calcium flux in response to ionomycin (5 μg/ml added at 25 seconds) was recorded on all available channels using an LSRII SORP flow cytometer. Cells were selectively gated on lymphocytes and singlets. The names assigned to the different detectors represent fluorochromes that are commonly detected in that channel. (A) Channels used to perform ratiometric analysis of Fura Red dye. (B) Representative examples of channels that display a shift in MFI following calcium mobilization. Channels such as these cannot be used for surface protein characterization in combination with Fura Red. (C) Channels with no change in MFI following calcium mobilization. Such channels are available for surface marker and cell viability analysis in combination with Fura Red dye. On the tested LSRII SORP, four available channels were identified, and fluorochromes commonly detected in these channels include: AlexFluor488, APC, APC-Cy7, and AlexaFluor700. One representative experiment of n = 2 shown.