Development of a dual microscope for simultaneous observation of oxygen distribution and blood flow in the microvascular bed

开发同时观察微血管床内氧气分布和血流的双显微镜

• 批准号: 05557007
• 负责人: ITOH Toshiyuki
• 金额: $ 4.74万
• 依托单位: Kyoto Prefectural University of Medicine
• 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
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-05557007/
• 关键词: microcirculation; microvascular bed; oxygen transport; oxygen tension; oxygen quenching; transport phenomenon; intravital microscopy; fluorescence microscopy; 拡散現像; 輸送現像

We developed an oxygen-quenching fluorescent membrane technique for in vivo visualization of pO2 distribution in the rat mesenteric microcirculation as optical patterns on the oxygen-sensitive membrane by epifluorescence microscopy. This membrane is highly gas-permeable to allow pO2 measurement, and is transparent enough to also permit observation of the microcirculation. By placing the oxygen-sensitive membrane in close proximity to the surface of mesentery without disturbing blood flow, it is possible to visualize the pO2 distribution in the microvascular network. We also developed a new microscope system for dual observation of pO2 distribution and microcirculation by using the oxygen-sensitive membrane technique.This new system is composed of two microscopes aligned face-to-face, a gastight and watertight chamber for the rat mesentery placed between two objectives, and a video recording system. One microscope (M1) equipped with a mercury lamp is employed for epi-illumination, and the other (M2) equipped with a halogen lamp for transillumination. A correcting diagonal mirror inserted in the optical path of M1 ensures proper direction of the two images. After -1 x1 mm area of the mesentery is selected through microscope M1 under transillumination, the light source is switched to the mercury lamp (B-excitation) for simultaneous observation of the fluorescence image through M1 and the microcirculation through M2. The recorded fluorescence image is digitized and converted to pO2 distribution by using Stern-Volmer relation, which can be displayd as 2-dimensional, 3-dimensional, or isobaric contour maps.The present system, which allows the simultaneous determination of pO2 distribution and microcirculatory parameters, should provide a novel approach to studies on the relationship between hemodynamics and oxygen transport dynamics under normal and abnormal conditions.

我们开发了一种氧猝灭荧光膜技术，用于通过落射荧光显微镜在氧敏感膜上的光学图案在大鼠肠系膜微循环中的pO 2分布的体内可视化。该膜具有高度透气性，可进行pO 2测量，并且足够透明，还可观察微循环。通过将氧敏感膜放置在肠系膜表面附近而不干扰血流，可以可视化微血管网络中的pO 2分布。我们还利用氧敏膜技术研制了一种新的显微镜系统，该系统由两个面对面的显微镜、置于两个物镜之间的气密水密的大鼠肠系膜室和一个视频记录系统组成，用于pO 2分布和微循环的双重观察。一个显微镜（M1）配备了汞灯用于落射照明，另一个（M2）配备了卤素灯用于透照。在M1的光路中插入的校正对角镜确保两个图像的正确方向。在透照下通过显微镜M1选择肠系膜的约1 × 1 mm区域后，将光源切换到汞灯（B激发），以同时观察通过M1的荧光图像和通过M2的微循环。利用Stern-Volmer关系，将荧光图像数字化后转换为pO 2分布图，可显示为二维、三维或等压等值线图，该系统可同时测定pO 2分布和微循环参数，为研究正常和异常情况下血流动力学与氧转运动力学之间的关系提供了一种新的方法。

项目成果

伊藤俊之 他: "蛍光法によるマイクロサ-キュレーションの計測" 非破壊生体計測研究連絡会ニュースレター. 16. 25-28 (1993)

Toshiyuki Ito 等：“使用荧光法测量微循环”无损生物仪器研究协调小组通讯。16. 25-28 (1993)

K.Yaegashi et al.: "A new method to visualize tissue oxygen distribution using fluorescent microscopy." Jpn.J.Physiol.43. S97 (1993)

K.Yaegashi 等人：“一种使用荧光显微镜可视化组织氧分布的新方法。”

上坂龍至 他: "生体組織における酸素輸送過程の二次元計測と解析" 生物物理. 33. S121 (1993)

Ryuji Uesaka 等人：“活组织中氧传输过程的二维测量和分析”生物物理学 33. S121 (1993)

K.Yaegashi et al.: "Oxygen transport in rat mesenteric microvessels visualized by an oxygensensitive membrane" In : "Progress in Microcirculation Research" (1994, ed.H.Niimi, M.Oda, T.Sawada, and R.-J.Xiu, Pergamon, Oxford, UK,pp.225-229.).

K.Yaegashi 等人：“通过氧敏感膜可视化大鼠肠系膜微血管中的氧传输”，见：“微循环研究进展”（1994 年，ed.H.Niimi、M.Oda、T.Sawada 和 R.-J）

K. Yaegashi et al.: "A new method to visualize tissue oxygen distribution using fluorescent microscopy." Jpn. J. Physiol.43. S97 (1993)

K. Yaegashi 等人：“一种使用荧光显微镜可视化组织氧分布的新方法。”