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.
Copyright © 2011 Elsevier B.V. All rights reserved.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Calcium / administration & dosage
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Calcium / pharmacology
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Capsaicin / pharmacology
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Electrophysiology / instrumentation*
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Equipment Design
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Exocytosis / drug effects
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Exocytosis / physiology*
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Fluorescent Dyes / analysis
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Glucose / pharmacology
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Heating / instrumentation*
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Indoles / analysis
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Ion Channel Gating / drug effects
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Ion Channel Gating / physiology*
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Islets of Langerhans / cytology
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Islets of Langerhans / drug effects
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Islets of Langerhans / physiology*
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Mice
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Mice, Inbred BALB C
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Micromanipulation / instrumentation
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Patch-Clamp Techniques / instrumentation
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Perfusion / instrumentation*
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Serotonin / analysis
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Single-Cell Analysis / instrumentation*
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Single-Cell Analysis / methods
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Solutions / administration & dosage*
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Solutions / pharmacology
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TRPV Cation Channels / drug effects
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TRPV Cation Channels / physiology*
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Temperature*
Substances
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Fluorescent Dyes
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Indoles
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Solutions
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TRPV Cation Channels
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TRPV1 protein, mouse
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Serotonin
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Glucose
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indo-1
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Capsaicin
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Calcium