Femnou Armel N, Giles Abigail, Balaban Robert S
Laboratory of Cardiac Energetics, National Heart, Lung, and Blood Institute, National Institutes of Health; Department of Biomedical Engineering, The George Washington University.
Laboratory of Cardiac Energetics, National Heart, Lung, and Blood Institute, National Institutes of Health.
J Vis Exp. 2019 May 12(147). doi: 10.3791/58992.
Absorbance spectroscopy of cardiac muscle provides non-destructive assessment of cytosolic and mitochondrial oxygenation via myoglobin and cytochrome absorbance respectively. In addition, numerous aspects of the mitochondrial metabolic status such as membrane potential and substrate entry can also be estimated. To perform cardiac wall transmission optical spectroscopy, a commercially available side-firing optical fiber catheter is placed in the left ventricle of the isolated perfused heart as a light source. Light passing through the heart wall is collected with an external optical fiber to perform optical spectroscopy of the heart in near real- time. The transmission approach avoids numerous surface scattering interference occurring in widely used reflection approaches. Changes in transmural absorbance spectra were deconvolved using a library of chromophore reference spectra, providing quantitative measures of all the known cardiac chromophores simultaneously. This spectral deconvolution approach eliminated intrinsic errors that may result from using common dual wavelength methods applied to overlapping absorbance spectra, as well as provided a quantitative evaluation of the goodness of fit. A custom program was designed for data acquisition and analysis, which permitted the investigator to monitor the metabolic state of the preparation during the experiment. These relatively simple additions to the standard heart perfusion system provide a unique insight into the metabolic state of the heart wall in addition to conventional measures of contraction, perfusion, and substrate/oxygen extraction.
心肌的吸收光谱分别通过肌红蛋白和细胞色素的吸光度对细胞质和线粒体氧合进行无损评估。此外,线粒体代谢状态的许多方面,如膜电位和底物进入,也可以进行估计。为了进行心脏壁透射光谱分析,将市售的侧射光纤导管作为光源置于离体灌注心脏的左心室中。穿过心脏壁的光由外部光纤收集,以近乎实时地对心脏进行光谱分析。这种透射方法避免了在广泛使用的反射方法中出现的大量表面散射干扰。使用发色团参考光谱库对透壁吸收光谱的变化进行解卷积,同时提供所有已知心脏发色团的定量测量。这种光谱解卷积方法消除了使用应用于重叠吸收光谱的常见双波长方法可能导致的固有误差,同时还提供了拟合优度的定量评估。设计了一个定制程序用于数据采集和分析,使研究人员能够在实验过程中监测制剂的代谢状态。除了收缩、灌注和底物/氧提取的传统测量方法外,对标准心脏灌注系统进行这些相对简单的补充,能够独特地洞察心脏壁的代谢状态。