Fluorescence Imaging of Electrical Activity in Cardiac Cells Using An All-Solid-State System

Emilia Entcheva, Yordan Kostov, Elko Tchernev, Leslie Tung

Research output: Contribution to journalArticle

Abstract

Tracking spatial and temporal determinants of cardiac arrhythmogenesis at the cellular level presents challenges to the optical mapping techniques employed. In this paper, we describe a compact system combining two nontraditional low-cost solutions for excitation light sources and emission filters in fluorescence measurements of transmembrane potentials, V m, or intracellular calcium, [Ca2+]i in cardiac cell networks. This is the first reported use of high-power blue and green light emitting diodes (LEDs), to excite cell monolayers stained with Vm- (di-8-ANEPPS) or [Ca2+]i - (Fluo-3) sensitive dyes. In addition, we use simple techniques for fabrication of suitable thin emission filters with uniform properties, no auto-fluorescence, high durability and good flexibility for imaging Vm or [Ca 2+]i. The battery-operated LEDs and the fabricated emission filters, integrated with a fiber-optic system for contact fluorescence imaging, were used as tools to characterize conduction velocity restitution at the macro-scale. The versatility of the LEDs for illumination is further emphasized through 1) demonstration of their usage for epi-illumination recordings at the single-cell level, and 2) demonstration of their unique high-frequency light modulation ability. The LEDs showed excellent stability as excitation light sources for fluorescence measurements; acceptable signal-to-noise ratio and negligible cell photodamage and indicator dye photobleaching were observed.

Original languageEnglish (US)
Pages (from-to)333-341
Number of pages9
JournalIEEE Transactions on Biomedical Engineering
Volume51
Issue number2
DOIs
StatePublished - Feb 1 2004

Keywords

  • Calcium
  • Cultured cardiac cells
  • LED excitation
  • Optical mapping
  • Voltage-sensitive dyes

ASJC Scopus subject areas

  • Biomedical Engineering

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