title: A fast solution switching system with temperature control for single cell measurements.
abstract: This article describes a perfusion system for biophysical single cell experiments at the physiological temperature. Our system regulates temperature of test solutions using a small heat exchanger that includes several capillaries. Water circulating inside the heat exchanger warms or cools test solutions flowing inside the capillaries. Temperature-controlled solutions are delivered directly to a single cell(s) through a multibarreled manifold that switches solutions bathing a cell in less than 1s. This solution exchange is optimal for patch clamp, single-cell microamperometry, and microfluorometry experiments. Using this system, we demonstrate that exocytosis from pancreatic β cells and activation of TRPV1 channels are temperature sensitive. We also discuss how to measure local temperature near a single cell under investigation.
authors.affiliation: Department of Physiology & Biophysics, University of Washington, Seattle, WA 98195-7290, USA. koh@u.washington.edu
authors.forename: Duk-Su
authors.initials: DS
authors.lastname: Koh
authors.forename: Liangyi
authors.initials: L
authors.lastname: Chen
authors.forename: Carmen A
authors.initials: CA
authors.lastname: Ufret-Vincenty
authors.forename: Seung-Ryoung
authors.initials: SR
authors.lastname: Jung
mesh_terms: D000818:Animals; D002118:Calcium / Q000008:administration & dosage / Q000494:pharmacology; D002211:Capsaicin / Q000494:pharmacology; D004594:Electrophysiology / Q000295:instrumentation*; D004867:Equipment Design; D005089:Exocytosis / Q000187:drug effects / Q000502:physiology*; D005456:Fluorescent Dyes / Q000032:analysis; D005947:Glucose / Q000494:pharmacology; D006361:Heating / Q000295:instrumentation*; D007211:Indoles / Q000032:analysis; D015640:Ion Channel Gating / Q000187:drug effects / Q000502:physiology*; D007515:Islets of Langerhans / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008846:Micromanipulation / Q000295:instrumentation; D018408:Patch-Clamp Techniques / Q000295:instrumentation; D010477:Perfusion / Q000295:instrumentation*; D012701:Serotonin / Q000032:analysis; D059010:Single-Cell Analysis / Q000295:instrumentation* / Q000379:methods; D012996:Solutions / Q000008:administration & dosage* / Q000494:pharmacology; D050916:TRPV Cation Channels / Q000187:drug effects / Q000502:physiology*; D013696:Temperature
chemical_list: D005456:Fluorescent Dyes; D007211:Indoles; D012996:Solutions; D050916:TRPV Cation Channels; C493921:TRPV1 protein, mouse; D012701:Serotonin; D005947:Glucose; C048960:indo-1; D002211:Capsaicin; D002118:Calcium