A fast solution switching system with temperature control for single cell measurements

J Neurosci Methods. 2011 Jul 15;199(1):35-42. doi: 10.1016/j.jneumeth.2011.04.023. Epub 2011 Apr 21.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / administration & dosage
  • Calcium / pharmacology
  • Capsaicin / pharmacology
  • Electrophysiology / instrumentation*
  • Equipment Design
  • Exocytosis / drug effects
  • Exocytosis / physiology*
  • Fluorescent Dyes / analysis
  • Glucose / pharmacology
  • Heating / instrumentation*
  • Indoles / analysis
  • Ion Channel Gating / drug effects
  • Ion Channel Gating / physiology*
  • Islets of Langerhans / cytology
  • Islets of Langerhans / drug effects
  • Islets of Langerhans / physiology*
  • Mice
  • Mice, Inbred BALB C
  • Micromanipulation / instrumentation
  • Patch-Clamp Techniques / instrumentation
  • Perfusion / instrumentation*
  • Serotonin / analysis
  • Single-Cell Analysis / instrumentation*
  • Single-Cell Analysis / methods
  • Solutions / administration & dosage*
  • Solutions / pharmacology
  • TRPV Cation Channels / drug effects
  • TRPV Cation Channels / physiology*
  • Temperature*

Substances

  • Fluorescent Dyes
  • Indoles
  • Solutions
  • TRPV Cation Channels
  • TRPV1 protein, mouse
  • Serotonin
  • Glucose
  • indo-1
  • Capsaicin
  • Calcium