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DEVELOPMENT OF INTRAVITAL LASER MICROSCOPE FOR THE OBSERVATION OF OXYGEN DISTRIBUTION

用于观察氧气分布的活体激光显微镜的研制

基本信息

批准号：
05558105
负责人：
KAMIYA Akira
金额：
$ 10.56万
依托单位：
UNIVERSITY OF TOKYO
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research (B)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

项目摘要

Although it has been recognized for over 100 years that one of the principal functions of the microcirculation is to deliver oxygen to tissue, our understanding of the processes involved in the transport of oxygen from blood to cells is incomplete. A major problem is that we do not know how the concentration of oxygen in tissue varies, particularly in the vicinity of blood vessels and around individual cells. In the present study we used newly designed phosphorescence intravital microscopy, which enable to measure oxygen tension (pO2) in small regions of tissue, to examine the gradient of pO2 in the tissue around the arteriole. Experiments were performed in anesthetized rat cremaster muscle. Pd-porphyrin complexes, the oxygen-dependent : phosphorescence dye were used to measure the quenching decay curve in the tissue. The pO2 was calculated from the time constant of the decay curve, following the Stern-Volmer equation : to/t=1+Kq・to・pO2 where to and are the phosphorescence lifetimes in the absence of O2 and in the tissue being measured and Kq is the quenching constant. Obtained arteriolar pO2 values decreased according to increase the number of branching (1st-order : 70mmHg, 2nd : 55mmHg, 3rd : 45mmHg), however venular pO2 values at the different position showed almost same level(30mmHg). Tissue pO2 values showed large gradient between the area closed to arterioles (arteriolar pO2 minus 15-20mmHg) and 100mum apart (5-8mmHg) . It was concluded that present method offers the continuous, non-invasive determina-tion of oxygen distribution in the skeletal muscle on a microscopic length scale.

虽然100多年来人们已经认识到微循环的主要功能之一是向组织输送氧气，但我们对氧气从血液到细胞的运输过程的了解还不完全。一个主要问题是，我们不知道组织中的氧浓度是如何变化的，特别是在血管附近和单个细胞周围。在本研究中，我们使用了新设计的磷光活体显微镜，它能够测量组织小区域的氧分压(PO2)，以检查小动脉周围组织的PO2梯度。实验在麻醉的大鼠提睾肌上进行。用氧依赖的磷光染料钯-卟啉络合物测定了组织中的猝灭衰减曲线。根据衰减曲线的时间常数计算pO2，遵循Stern-Volmer方程：to/t=1+kq·to·pO2，其中to和是在无氧和被测组织中的磷光寿命，kq是猝灭常数。动脉血氧分压(PO2)值随动脉分支数目的增加而降低(1级：70 mm Hg，2级：55 mm Hg，3级：45 mm Hg)，而不同部位的静脉PO2值基本相同(30 Mm Hg)。组织氧分压显示接近小动脉的区域(动脉血氧分压减15-20毫米汞柱)与相隔100微米(5-8毫米汞柱)之间有很大的梯度。结论：本方法提供了在显微长度尺度上连续、非侵入性地测定骨骼肌氧分布的方法。