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Automated Method for Tracking Vast Numbers of FITC‐Labeled RBCs in Microvessels of Rat Brain In Vivo Using a High‐Speed Confocal Microscope System
Author(s) -
Tomita Minoru,
Osada Takashi,
Schiszler Istvan,
Tomita Yutaka,
Unekawa Miyuki,
Toriumi Haruki,
Tanahashi Norio,
Suzuki Norihiro
Publication year - 2008
Publication title -
microcirculation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.793
H-Index - 83
eISSN - 1549-8719
pISSN - 1073-9688
DOI - 10.1080/10739680701567089
Subject(s) - confocal , microscope , biomedical engineering , confocal microscopy , in vivo , fluorescence microscope , frame rate , artificial intelligence , computer science , materials science , computer vision , chemistry , fluorescence , optics , physics , biology , medicine , microbiology and biotechnology
High‐speed camera investigation of rapidly moving red blood cells (RBCs) in the microvasculature has been limited by an inability to handle the vast volume of data. We have developed a novel method to analyze large numbers of RBC images captured by a high‐resolution, high‐speed camera fitted on a confocal fluorescence microscope, to determine the velocities of individual RBCs in capillaries in vivo . Fluorescein isothiocyanate (FITC)‐labeled RBCs flowing in the microvasculature of the cerebral cortex of urethane‐anesthetized Wistar rats were recorded through the skull window on video clips during specified periods at high frame rates (500 fps). Sequential frames of moving RBCs in the video clips for a specified period were analyzed offline with in‐house software (KEIO‐IS2). Images of RBCs acquired were numbered automatically in order of appearance and displayed in a two‐dimensional (2‐D) RBC tracking map. The velocities of individual RBCs were automatically computed based on the RBC displacement per frame multiplied by the frame rate (fps), and the results were displayed in a 2‐D velocity map and a 2‐D RBC number map. Single capillaries were identified by staining with FITC‐dextran. The mean capillary velocity of RBCs was evaluated as 2.05 ± 1.59 mm/second in video clips obtained at 500 fps. This method is considered to have wide potential applicability.