Publication detail

Tissue Response during Staining and Illumination of Voltage-Sensitive Dye in Rabbit Myocardium

NOVÁKOVÁ, M. NOGOVÁ, K. PROVAZNÍK, I. BARDOŇOVÁ, J.

Original Title

Tissue Response during Staining and Illumination of Voltage-Sensitive Dye in Rabbit Myocardium

English Title

Tissue Response during Staining and Illumination of Voltage-Sensitive Dye in Rabbit Myocardium

Type

conference paper

Language

en

Original Abstract

Voltage-sensitive dyes (VSD's) undergo changes in their electronic structure, and consequently their fluorescence spectra, in response to changes in the surrounding electric field and are therefore used for recording of monophasic action potentials (MAPs) by optical method in animal hearts. VSD di-4-ANEPPS is employed in our laboratory to record MAPs in isolated rabbit hearts perfused according to Langendorff (80mmHg) with Krebs-Henseleit solution (K-H, 37C, 1, 25mM Ca2+). After stabilization (30min), the tissue is stained with di-4-ANEPPS (2mM) diluted in K-H (20-25min) and then washed-out with dye-free K-H (20-25min). Then measurement of MAPs is performed. So called photodynamic damage or phototoxic effects of VSDs have been reported under intense illumination, with alteration of electrical activity of the spontaneously beating heart. In order to determine whether this side effect is present in our experimental model, we followed electrogram, mean coronary flow and formation of hydroxyl radicals in the control, staining and washout periods and during illumination (20min). The hearts then underwent examination on electron microscopy to detect possible ultrastructural changes. Only minor changes of electrogram were observed during staining and washout (changes in T wave morphology and episodic atrio-ventricular dissociation). The heart rate decreased slightly in staining, partially restored in washout and did not changed subsequently. No such changes were detected under focal illumination and the optical signal did not show any signs of fading. No significant changes were observed in mean coronary flow during the whole experiment, no hydroxyl radicals were formed and no ultrastructural changes were found. We conclude that rabbit myocardium is resistant to di-4-ANEPPS side effects during staining and also resistant to bleaching during intense illumination.

English abstract

Voltage-sensitive dyes (VSD's) undergo changes in their electronic structure, and consequently their fluorescence spectra, in response to changes in the surrounding electric field and are therefore used for recording of monophasic action potentials (MAPs) by optical method in animal hearts. VSD di-4-ANEPPS is employed in our laboratory to record MAPs in isolated rabbit hearts perfused according to Langendorff (80mmHg) with Krebs-Henseleit solution (K-H, 37C, 1, 25mM Ca2+). After stabilization (30min), the tissue is stained with di-4-ANEPPS (2mM) diluted in K-H (20-25min) and then washed-out with dye-free K-H (20-25min). Then measurement of MAPs is performed. So called photodynamic damage or phototoxic effects of VSDs have been reported under intense illumination, with alteration of electrical activity of the spontaneously beating heart. In order to determine whether this side effect is present in our experimental model, we followed electrogram, mean coronary flow and formation of hydroxyl radicals in the control, staining and washout periods and during illumination (20min). The hearts then underwent examination on electron microscopy to detect possible ultrastructural changes. Only minor changes of electrogram were observed during staining and washout (changes in T wave morphology and episodic atrio-ventricular dissociation). The heart rate decreased slightly in staining, partially restored in washout and did not changed subsequently. No such changes were detected under focal illumination and the optical signal did not show any signs of fading. No significant changes were observed in mean coronary flow during the whole experiment, no hydroxyl radicals were formed and no ultrastructural changes were found. We conclude that rabbit myocardium is resistant to di-4-ANEPPS side effects during staining and also resistant to bleaching during intense illumination.

Keywords

tissue, voltage-sensitive dye, rabbit, heart

RIV year

2007

Released

01.12.2007

Publisher

IEEE

Location

Durham, USA

ISBN

978-1-4244-2533-4

Book

Computers in Cardiology

Pages from

665

Pages to

668

Pages count

4

BibTex


@inproceedings{BUT27952,
  author="Marie {Nováková} and Kateřina {Nogová} and Ivo {Provazník} and Jana {Kolářová}",
  title="Tissue Response during Staining and Illumination of Voltage-Sensitive Dye in Rabbit Myocardium",
  annote="Voltage-sensitive dyes (VSD's) undergo changes in their electronic structure, and consequently their fluorescence spectra, in response to changes in the surrounding electric field and are therefore used for recording of monophasic action potentials (MAPs) by optical method in animal hearts. VSD di-4-ANEPPS is employed in our laboratory to record MAPs in isolated rabbit hearts perfused according to Langendorff (80mmHg) with Krebs-Henseleit solution (K-H, 37C, 1, 25mM Ca2+). After stabilization (30min), the tissue is stained with di-4-ANEPPS (2mM) diluted in K-H (20-25min) and then washed-out with dye-free K-H (20-25min). Then measurement of MAPs is performed. So called photodynamic damage or phototoxic effects of VSDs have been reported under intense illumination, with alteration of electrical activity of the spontaneously beating heart. In order to determine whether this side effect is present in our experimental model, we followed electrogram, mean coronary flow and formation of hydroxyl radicals in the control, staining and washout periods and during illumination (20min). The hearts then underwent examination on electron microscopy to detect possible ultrastructural changes. Only minor changes of electrogram were observed during staining and washout (changes in T wave morphology and episodic atrio-ventricular dissociation). The heart rate decreased slightly in staining, partially restored in washout and did not changed subsequently. No such changes were detected under focal illumination and the optical signal did not show any signs of fading. No significant changes were observed in mean coronary flow during the whole experiment, no hydroxyl radicals were formed and no ultrastructural changes were found. We conclude that rabbit myocardium is resistant to di-4-ANEPPS side effects during staining and also resistant to bleaching during intense illumination.",
  address="IEEE",
  booktitle="Computers in Cardiology",
  chapter="27952",
  institution="IEEE",
  year="2007",
  month="december",
  pages="665--668",
  publisher="IEEE",
  type="conference paper"
}