Multiple injury approach and its use for toxicity studies.
We present an experimental approach that allows exposure of cells plated on a single coverslip to multiple distinct environments. The original chamber design created a small region of injury using geometrically defined flows of the control and ischemic solutions. Modifications of the original chamber design presented in this article produce a range of flow patterns that can be advantageous for a variety of imaging applications. These applications include: experiments that address effects of different treatments applied to a cell network, parallel testing of negative and positive controls using a single coverslip, border effect studies, evaluation of the treatment's reversibility, and simultaneous monitoring of a cell layer loaded with different fluorescent indicators. The method also can be used to reveal both micro- and macroscopic features of propagation, conduction, and cell coupling in a normal or altered cardiac cell network. These possibilities are illustrated in cultures of neonatal rat cardiomyocytes using oxidant- and calcium-sensitive fluorescent indicators.
Arutunyan
Ara
A
Department of Physiology, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA.
Swift
Luther
L
Sarvazyan
Narine
N
D000818:Animals; D001145:Arrhythmias, Cardiac / Q000503:physiopathology; D002448:Cell Adhesion / Q000502:physiology; D002478:Cells, Cultured; D005456:Fluorescent Dyes; D015428:Myocardial Reperfusion Injury / Q000473:pathology* / Q000503:physiopathology; D032383:Myocytes, Cardiac / Q000473:pathology*; D010477:Perfusion; D051381:Rats; D017207:Rats, Sprague-Dawley; D012008:Recurrence; D014116:Toxicology / Q000379:methods*
D005456:Fluorescent Dyes