The use of flow cytometry to examine calcium signalling by TRPV1 in mixed cell populations

Flow cytometric analysis of calcium mobilisation has been in use for many years in the study of specific receptor engagement or isolated cell:cell communication. However, calcium mobilisation/signaling is key to many cell functions including apoptosis, mobility and immune responses. Here we combine multiplex surface staining of whole spleen with Indo-1 AM to visualise calcium mobilisation and examine calcium signaling in a mixed immune cell culture over time. We demonstrate responses to a TRPV1 agonist in distinct cell subtypes without the need for cell separation. Multi parameter staining alongside Indo-1 AM to demonstrate calcium mobilization allows the study of real time calcium signaling in a complex environment. •This paper aims to help discuss calcium signaling in TRPV1+ splenocytes via flow cytomerty.•We re-introduce a previously used technique to help identify different cell populations accurately with adequate sensitivity.•This technique is economical and easy to reproduce.