Microhemodynamics after Infusion of an Artificial O_2 Carriers Using Laser Flash Irradiation.

（1）血红蛋白囊泡（HbV）已被开发以替代RBC的功能。我们开发了新的方法来分析输注HbV后啮齿动物的微血流动力学，并评估HbV的安全性和有效性。将标记的HbV注入仓鼠体内，用激光共聚焦显微镜原位观察肝脏。这允许可视化在窦状隙中流动的HbV，并且颗粒最终被捕获并积聚在枯否细胞中，这可以被观察为增强的荧光。使用抗人Hb抗体和血清临床化学的组织学分析表明，HbV的迅速代谢和排泄没有任何不可逆的变化。（2）采用无创性经皮闪光照明技术，在显微镜下观察了白蛋白极度血液稀释时大鼠尾部皮肤毛细血管内红细胞的流动。在60%水平下，RBC流速单调增加至基础值的294 ± 110%。 ...更多信息 血液交换，与腹主动脉血流量相比增加到148 ± 20%。毛细血管红细胞压积的下降比例似乎与全身红细胞压积不同，这与先前的观察结果一致。这些结果表明，在血液稀释过程中的微血流动力学特性可以transmitted分析与这种新技术，而无需手术准备的皮肤。（3）用内径为28 μm的透氧聚合物管在缺氧环境中检测HbVs的氧释放。用倒置显微镜和带光子计数检测器的扫描光栅分光光度计组成的装置测量O_2释放，根据Hb在Q带的可见吸收光谱确定O_2释放速率。HbV/RBC混合物的O_2释放速率与单独RBC的O_2释放速率相似。另一方面，添加脱细胞血红蛋白溶液的红细胞显着提高脱氧速率。这主要是由于颗粒尺寸的差异（250与7 nm）影响了它们的扩散以促进O_2的传输。少

(1)Hemoglobin-vesicles (HbV) has been developed to substitute the function of RBCs. We developed new methods to analyze the microhemodynamics after infusion of HbV into rodents, and evaluated the safety and efficacy of HbV. Fluorescence-labeled HbV was infused into hamsters and the liver was observed in situ with a laser con focal microscopy. This allowed visualization of HbV flowing in the sinusoid, and the particles were finally captured and accumulated in the Kupffer cells, which could be observed as intensified fluorescence. Histopathological analysis using anti-human Hb antibody and serum clinical chemistry demonstrated the prompt metabolism and excretion of HbV without any irreversible changes.(2)Capillary RBC flow in the skin of rat tail during an extreme hemodilution with albumin was microscopically observed with a noninvasive transcutaneous technique using flash epi-illumination. The RBC flow rates monotonously increased up to 294 ± 110% of the basal value at the 60% level of … More blood exchange, which was in contrast to the abdominal aortic blood flow increase to 148 ± 20%. Capillary hematocrit did not appear to fall in the same proportion as the systemic hematocrit, which coincided with the previous observations. These results indicate that microhemodynamic properties during hemodilution can be transcutaneously analyzed with this new technique without surgical preparation of the skin.(3)The O_2 release from flowing HbVs was examined using an O_2-permeable polymer tube (inner diameter, 28 μm) exposed to a deoxygenated environment. Measurement of O_2 release was performed using an apparatus that consisted of an inverted microscope and a scanning-grating spectrophotometer with a photon-count detector, and the rate of O_2 release was determined based on the visible absorption spectrum in the Q band of Hb. The rate of O_2 release from the HbV/RBC mixtures was similar to that of RBCs alone. On the other hand, the addition of acellular Hb solution to RBCs significantly enhanced the rate of deoxygenation. This should mainly be due to the difference in the particle size (250 vs. 7 nm) that affects their diffusion for the facilitated O_2 transport. Less