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首页> 外文期刊>Computer Methods and Programs in Biomedicine: An International Journal Devoted to the Development, Implementation and Exchange of Computing Methodology and Software Systems in Biomedical Research and Medical Practice >Validation of automated spectrofluroimetry for measurement of regional organ perfusion using fluorescent microspheres.
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Validation of automated spectrofluroimetry for measurement of regional organ perfusion using fluorescent microspheres.

机译：验证了使用荧光微球的自动分光光度法测量区域器官灌注的有效性。

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The fluorescent microsphere (FM) method for determination of regional organ blood flow is labor intensive, requiring processing of 100 s to 1000 s of samples per organ. To save time and reduce handling errors, we developed an automated fluorescence analysis system by interfacing a commercially available spectrofluorimeter and sample delivery unit to a PC, operated by a Windows95(R)-based program, WINFAC. WINFAC allows versatile analysis setup and instrument control for fluorescent intensity acquisition at fixed wavelengths or by synchronous scanning. Data is presented on screen, in real-time, and stored in text format. Reference analyses are performed at specified intervals and the coefficient of variation is continuously updated to monitor instrument performance. The automated system was validated against radioactive microspheres (RM) for lung perfusion in a pig and sheep and evaluated for analysis reproducibility. Fluorescent and radioactive microspheres were delivered by simultaneous injection into a femoral vein. Lungs were excised, flushed, dried at total lung capacity, and cubed into approximately 2cm(3) pieces (n=833 and 1560, pig and sheep, respectively). Radioactive count rates were determined for each lung piece (corrected for decay, background and spillover). Fluorescent dyes were extracted in solvent and intensities were determined at fixed wavelengths, using the automated spectrofluorimeter (corrected for background and spillover). Multi-color reference solutions were measured every 50 samples to monitor instrument reproducibility. Blood flow estimates for each piece determined by RM and FM methods were highly correlated: R(2)=0.98+/-0.017, Slopes=1.00+/-0.007 and Intercepts=0.00+/-0.006 (mean+/-SD). The CV of repeat reference analyses was 0.71%+/-0.16, a 30% to 50% reduction relative to manual analysis. Automated spectrofluorimetry reduces measurement errors and is a reliable and time saving advancement. With this technology, use of FM to measure regional lung perfusion approaches the ease and accuracy of the RM method.
机译：用于确定局部器官血流的荧光微球（FM）方法需要大量劳动，每个器官需要处理100 s至1000 s的样品。为了节省时间并减少处理错误，我们通过将市售的荧光分光光度计和样品输送装置连接到PC上，开发了一种自动荧光分析系统，该PC由基于Windows95（R）的程序WINFAC操作。 WINFAC允许进行多种分析设置和仪器控制，以固定波长或通过同步扫描来采集荧光强度。数据实时显示在屏幕上，并以文本格式存储。参考分析以指定的时间间隔执行，并且变异系数会不断更新以监控仪器性能。该自动化系统针对放射性微球（RM）进行了猪和绵羊肺灌注的验证，并评估了分析的可重复性。通过同时注射入股静脉来递送荧光和放射性微球。肺被切除，冲洗，在肺总容量下干燥，并切成约2cm（3）的小块（分别为n = 833和1560，猪和羊）。确定每个肺片的放射性计数率（针对衰变，背景和溢出进行校正）。使用自动荧光分光光度计（针对背景和溢出进行校正），在溶剂中提取荧光染料，并在固定波长下确定强度。每50个样品测量一次多色参比溶液，以监测仪器的重现性。通过RM和FM方法确定的每件血流估计值高度相关：R（2）= 0.98 +/- 0.017，斜率= 1.00 +/- 0.007和截距= 0.00 +/- 0.006（平均值+/- SD）。重复参考分析的CV为0.71％+ /-0.16，相对于手动分析降低了30％至50％。自动化的荧光分光光度法减少了测量误差，是一种可靠且节省时间的进步。借助这项技术，使用FM来测量局部肺灌注可以接近RM方法的简便性和准确性。

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