Scalable multi-parametric imaging of excitable tissue

Thesis

The field of cardiac electrophysiological imaging has advanced tremendously in the past three decades with developments in fluorescent dyes, photodetectors, optical filters, illumination sources, computers and electronics. This thesis describes several scalable multi-parametric imaging systems and their application to cardiac tissue preparations at various levels of complexity. Using off-the-shelf components, single-camera multi-parametric optical mapping systems are described for various fluorescent dye combinations and single-element photodiode-based fibre-optic detection systems are described for drug-testing applications. The instruments described take advantage of modern voltage-sensitive dyes, multi-band optical filters and powerful light-emitting-diodes, from the ultraviolet to the red.

The two electrophysiological parameters focused on were transmembrane voltage and the intracellular calcium concentration. Several voltage and calcium dye combinations were established, which produce no signal cross-talk. Furthermore, second- and third-generation voltage dyes were characterized in cardiac tissue, in vitro and in vivo.

The developed systems were then applied to isolated Langendorff-perfused whole-hearts, in vivo whole-hearts, thin ventricular tissue-slices and human induced pluripotent stem cell-derived cardiac tissue. The interventions applied include accurately-timed electrical and mechanical local stimulation of the whole-heart to generate ectopic beats, cardiotoxic drugs and flash-photolysis of caged-compounds.

With the high-throughput demands of drug discovery and testing, further development of scalable optical electrophysiological systems may prove critical in reducing attrition and costs. And for in vivo optical mapping, development of minimally-invasive and clinically-relevant optical systems will be essential in validating existing theories based on in vitro experiments and exploring cardiac function and behaviour with the heart intact in the organism.

Authors: Lee, P

• Alternative title: cardiac imaging
• Publication date: 2012
• Type of award: DPhil
• Level of award: Doctoral
• Awarding institution: Oxford University, UK
• Language: English
• Keywords: fluorescence; electrophysiology; multi-parametric; imaging; cardiac
• Subjects: Biophysics