title: A modular perfused chamber for low- and normal-temperature microscopy of living cells.
abstract: An inexpensive modular perfused chamber (MPC) designed for low- and normal-temperature live-cell imaging is presented. The device consists of four lathed pieces of stainless steel assembled as a cylindrical open chamber that can hold either round or square glass coverslips. The chamber is connected to a thermal-bath operating with recirculation. For image acquisition at 4°C, cooled air is blown toward the coverslip surface to prevent condensation. Principal advantages of this device are thermal stability in the sample environment, rapid response to changes in temperature set point, and easy sample insertion. The device enables the study of dynamic processes in cells governed by large temperature differences such as those imposed by hypothermic preservation of cells (0-4°C) followed by rewarming to normothermia (37°C). The capabilities of the MPC were demonstrated by monitoring the internalization of fluorescent quantum dots (QDs) in rat hepatocytes after hypothermic storage and during rewarming with an inverted microscope.
authors.affiliation: Centro Binacional (Argentina-Italia) de Investigaciones en Criobiología Clínica y Aplicada, Universidad Nacional de Rosario, Rosario, Argentina. vsigot@fbioyf.unr.edu.ar
authors.forename: Valeria
authors.initials: V
authors.lastname: Sigot
authors.forename: José Manuel
authors.initials: JM
authors.lastname: Pellegrino
authors.forename: Edgardo Elvio
authors.initials: EE
authors.lastname: Guibert
authors.forename: Joaquín Valentín
authors.initials: JV
authors.lastname: Rodriguez
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation*; D003080:Cold Temperature; D003584:Cytological Techniques / Q000295:instrumentation*; D004867:Equipment Design; D005456:Fluorescent Dyes / Q000737:chemistry / Q000493:pharmacokinetics; D022781:Hepatocytes / Q000737:chemistry / Q000378:metabolism; D008853:Microscopy / Q000295:instrumentation* / Q000379:methods; D045663:Quantum Dots; D051381:Rats; D017208:Rats, Wistar
chemical_list: D005456:Fluorescent Dyes