title: A microfluidic device to sort cells based on dynamic response to a stimulus.
abstract: Single cell techniques permit the analysis of cellular properties that are obscured by studying the average behavior of cell populations. One way to determine how gene expression contributes to phenotypic differences among cells is to combine functional analysis with transcriptional profiling of single cells. Here we describe a microfluidic device for monitoring the responses of single cells to a ligand and then collecting cells of interest for transcriptional profiling or other assays. As a test, cells from the olfactory epithelium of zebrafish were screened by calcium imaging to identify sensory neurons that were responsive to the odorant L-lysine. Single cells were subsequently recovered for transcriptional profiling by qRT-PCR. Responsive cells all expressed TRPC2 but not OMP, consistent with known properties of amino-acid sensitive olfactory neurons. The device can be adapted for other areas in biology where there is a need to sort and analyze cells based on their signaling responses.
authors.affiliation: Microfluidics Systems Biology Lab, Institute of Molecular and Cell Biology, Singapore, Singapore.
authors.forename: Swee Jin
authors.initials: SJ
authors.lastname: Tan
authors.forename: Michelle Z L
authors.initials: MZ
authors.lastname: Kee
authors.forename: Ajay Sriram
authors.initials: AS
authors.lastname: Mathuru
authors.forename: William F
authors.initials: WF
authors.lastname: Burkholder
authors.forename: Suresh J
authors.initials: SJ
authors.lastname: Jesuthasan
mesh_terms: D000818:Animals; D002118:Calcium / Q000737:chemistry; D058948:Cell Tracking / Q000379:methods*; D015870:Gene Expression / Q000235:genetics; D008024:Ligands; D008239:Lysine / Q000008:administration & dosage; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D009831:Olfactory Mucosa / Q000187:drug effects / Q000378:metabolism; D018034:Olfactory Receptor Neurons / Q000187:drug effects / Q000378:metabolism*; D011984:Sensory Receptor Cells / Q000187:drug effects; D059010:Single-Cell Analysis / Q000295:instrumentation* / Q000379:methods; D015027:Zebrafish
chemical_list: D008024:Ligands; D008239:Lysine; D002118:Calcium