Cardiovascular disease, especially coronary heart disease (CHD) caused by coronary atherosclerosis, has been a leading cause of death world-wide. Traditional methods of diagnosing CHD, including coronary CT angiography and histopathological examination, are limited either by the low image resolution or time-consuming sample preparation. Spectrum- and time-resolved multiphoton microscopy has the potential to circumvent this problem because it can not only interrogate the three-dimensional structure of coronary arterial intima but also reveal its biochemical components at subcellular resolution. However, its clinical utility hasn’t been thoroughly assessed. Here, we adopted a spectrum- and timeresolved multiphoton microscope to characterize unstained human coronary atherosclerotic plaque ex vivo. According to the fluorescence spectra, the collagen fibers and elastic fibers in the coronary arterial intima can be separated clearly. We also analyzed the changes in the relative content of collagen fibers and elastic fibers by calculating the ratio of these two kinds of fibers in normal and diseased coronary arterial wall and found that the coronary atherosclerotic plaque regions has a higher ratio compared to that of normal tissue. Finally, we assessed the biochemical variations in coronary atherosclerotic tissues by measuring the fluorescence lifetime and found a lower mean fluorescence lifetime in coronary atherosclerotic plaque. This study indicates that the spectrum- and time-resolved multiphoton microscopy has the potential to distinguish atherosclerotic plaque on the coronary arterial wall from normal tissues.
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