Large-scale investigation of the olfactory receptor space using a microfluidic microwell array.
The mammalian olfactory system is able to discriminate among tens of thousands of odorant molecules. In mice, each odorant is sensed by a small subset of the approximately 1000 odorant receptor (OR) types, with one OR gene expressed by each olfactory sensory neuron (OSN). However, the sum of the large repertoire of OR-OSN types and difficulties with heterologous expression have made it almost impossible to analyze odorant-responsiveness across all OR-OSN types. We have developed a microfluidic approach that allowed us to screen over 20,000 single cells at once in microwells. By using calcium imaging, we were able to detect and analyze odorant responses of about 2900 OSNs simultaneously. Importantly, this technique allows for both the detection of rare responding OSNs as well as the identification of OSN populations broadly responsive to odorants of unrelated structures. This technique is generally applicable for screening large numbers of single cells and should help to characterize rare cell behaviors in fields such as toxicology, pharmacology, and cancer research.
Figueroa
Xavier A
XA
Department of Bioengineering, University of Washington, Box 355061, Seattle, WA 98195, USA.
Cooksey
Gregory A
GA
Votaw
Scott V
SV
Horowitz
Lisa F
LF
Folch
Albert
A
D001681:Biological Assay / Q000295:instrumentation*; D002469:Cell Separation / Q000295:instrumentation*; D002478:Cells, Cultured; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006801:Humans; D046228:Microarray Analysis / Q000295:instrumentation*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D009812:Odorants; D018034:Olfactory Receptor Neurons / Q000187:drug effects / Q000502:physiology*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012903:Smell / Q000187:drug effects / Q000502:physiology*
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