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Biophys J, January 2001, p. 516-530, Vol. 80, No. 1
Department of Pharmacology, SUNY Health Science Center, Syracuse, New York 13210 USA
Voltage-sensitive fluorescent dyes have become powerful
tools for the visualization of excitation propagation in the heart. However, until recently they were used exclusively for surface recordings. Here we demonstrate the possibility of visualizing the
electrical activity from inside cardiac muscle via fluorescence measurements in the transillumination mode (in which the light source
and photodetector are on opposite sides of the preparation). This mode
enables the detection of light escaping from layers deep within the
tissue. Experiments were conducted in perfused (8 mm thick) slabs of
sheep right ventricular wall stained with the voltage-sensitive dye
di-4-ANEPPS. Although the amplitude and signal-to-noise ratio recorded
in the transillumination mode were significantly smaller than those
recorded in the epi-illumination mode, they were sufficient to reliably
determine the activation sequence. Penetration depths (spatial decay
constants) derived from measurements of light attenuation in cardiac
muscle were 0.8 mm for excitation (520 ± 30 nm) and 1.3 mm for
emission wavelengths (640 ± 50 nm). Estimates of emitted
fluorescence based on these attenuation values in 8-mm-thick tissue
suggest that 90% of the transillumination signal originates from a
4-mm-thick layer near the illuminated surface. A 69% fraction of the
recorded signal originates from 
1 mm below the surface.
Transillumination recordings may be combined with endocardial and
epicardial surface recordings to obtain information about
three-dimensional propagation in the thickness of the myocardial wall.
We show an example in which transillumination reveals an intramural
reentry, undetectable in surface recordings.
Biophys J, January 2001, p. 516-530, Vol. 80, No. 1
© 2001 by the Biophysical Society 0006-3495/01/01/516/15 $2.00
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