Abstract

Introduction: Imaging the coronary microcirculation in the beating heart is challenging due to motion artifact from cardiac contraction and respiration, which limits the application of high-resolution microscopy techniques. In the brain and other tissues with less motion artifact, optical coherence tomography angiography (OCTA) has been successfully used in rodents to create label-free maps of blood flow down to the capillary level. We hypothesized that advanced cardiac and respiratory gating together with high-speed optical coherence tomography can be used to implement OCTA of the capillary microcirculation in vivo in the mouse heart. Methods: C57BL/6 mice (n=30) were imaged through a sternotomy while anesthetized and ventilated. OCTA was performed using a custom tissue stabilizer and a Thorlabs TEL 321 Spectral Domain OCT system, which was synchronized to physiologic signals using custom hardware and software and cardiac pacing protocols. The system had an optical center wavelength of 1300 nm and produced axial (depth) and lateral resolutions of 4.2 μm and 13 μm, respectively. Angiograms were created from the difference between two consecutive images acquired during diastole, and a volumetric three-dimensional scan was acquired over multiple cardiac cycles. Images were analyzed in MATLAB and ImageJ to produce projection angiograms of the coronary microcirculation. Results: OCTA with cardiac and respiratory gating can produce high-resolution angiograms of the coronary microcirculation without exogenous contrast agents. OCTA has improved imaging depth and field of view compared to conventional confocal microscopy, and can produce widefield coronary angiograms (Fig. 1A) while maintaining the ability to visualize individual capillaries (Fig. 1B). Conclusion: In vivo OCTA can image the coronary microcirculation in the heart and will provide a powerful new tool for investigating coronary microvascular dysfunction in mouse models of heart disease.

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