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Local Field Fluorescence Microscopy: Imaging Cellular Signals in Intact Hearts
Yuriana Aguilar-Sanchez1, Diego Fainstein2, Rafael Mejia-Alvarez3
1School of Natural Sciences, University of California, Merced.
Journal of Visualized Experiments : Jove
|April 1, 2017
Summary
Local Field Fluorescence Microscopy (LFFM) enables cellular signal measurement in intact hearts, advancing cardiovascular physiology research. This technique offers new insights into heart function under normal and pathological conditions.
Area of Science:
- Cardiovascular Physiology
- Biophysics
- Optical Microscopy
Background:
- Molecular signaling studies in the heart typically use isolated myocytes.
- Pathological conditions like ischemia and arrhythmias require whole-organ level understanding.
Purpose of the Study:
- To introduce Local Field Fluorescence Microscopy (LFFM) for measuring cellular signals in intact hearts.
- To demonstrate LFFM's applicability in cardiovascular physiology.
Main Methods:
- Utilized a Langendorff-perfused heart system combined with optical fibers.
- Employed three LFFM epifluorescence arrangements: pulsed LFFM (PLFFM), continuous wave LFFM (CLFFM), and pulsed LED fluorescence microscopy (PLEDFM).
- Monitored cardiac variables like cytosolic [Ca2+], intra-sarcoplasmic reticulum [Ca2+], and membrane potentials using fluorescent indicators.
Main Results:
- Successfully demonstrated LFFM for in-situ cellular signal measurement in the intact heart.
- Showcased the versatility of LFFM with different light sources (laser pulses, continuous laser, LEDs) and modulation techniques.
- Validated the monitoring of key physiological parameters within the whole organ context.
Conclusions:
- LFFM is a powerful spectroscopic technique for studying the intact heart at the cellular level.
- This method provides a valuable tool for investigating cardiovascular physiology under both normal and pathological conditions.
- LFFM opens new avenues for understanding complex cardiac diseases.