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      Off-Line Analysis of Red Blood Cell Velocity in Renal Arterioles

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          Abstract

          Videomicroscopic methods with off-line analysis of microcirculatory parameters by multifunctional computer-assisted image analysis systems have significant advantages for in vivo microvascular research. A limitation of these methods is, however, that red blood cell velocities (V<sub>RBC</sub>) exceeding 2 mm/s cannot be measured using standard video framing rates. In the present study, a high-speed video camera, recording up to 600 frames per second, was incorporated in the set-up, and V<sub>RBC</sub> was measured off-line with the line-shift-diagram method. The aim of this study was to test the reproducibility and validity of the method using a high-speed video camera and to evaluate its applicability in vivo. V<sub>RBC</sub> were measured in arterioles of the split hydronephrotic kidney. The intra- and interindividual variability was small for V<sub>RBC</sub> below 40 mm/s. The validity of the method was tested using the mass conservation principle and found to be at least as good as that of the dual-slit photometric technique. The present approach extends the application of videomicroscopy coupled to image analysis systems to the analysis of high V<sub>RBC</sub>.

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          Determination of Red Blood Cell Velocity by Video Shuttering and Image Analysis

          A novel modification of conventional video imaging techniques has been developed to determine the velocity of red blood cells (RBCs), which offers compatibility with existing video-based methods for determining blood oxygenation and hemoglobin concentration. Traditional frame-by-frame analysis of video recordings limits the maximum velocity that can be measured for individual cells in vivo to about 2 mm/s. We have extended this range to about 20 mm/s, by electronic shuttering of an intensified charge-coupled device camera to produce multiple images of a single RBC in the same video frame. RBCs were labeled with fluorescein isothiocyanate and the labeled cells (FRBCs) were used as probes to determine RBC velocities in microvessels of the hamster retractor muscle. Velocity was computed as the product of the distance between centroids of two consecutive image positions of a FRBC and the shuttering frequency of the camera intensifier. In vitro calibrations of the system using FRBC and Sephadex beads coated onto a rotating disk yielded an average coefficient of variation of about 6%. Flow conservation studies at bifurcations indicated that the maximum diameter of microvessels below which all the FRBCs in the lumen could be detected was 50 microm. The technique was used to estimate mean-FRBC velocity distributions in vessels with diameters ranging from 8 to 50 microm. The mean-FRBC velocity profiles were found to be blunter than would be expected for Poiseuille flow. Single FRBCs tracked along an unbranched arteriole exhibited significant temporal variations in velocity.
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            Author and article information

            Journal
            JVR
            J Vasc Res
            10.1159/issn.1018-1172
            Journal of Vascular Research
            S. Karger AG
            1018-1172
            1423-0135
            2000
            February 2000
            07 March 2000
            : 37
            : 1
            : 26-31
            Affiliations
            aRenal Unit, Department of Internal Medicine, University Hospital, Gent, Belgium, and bServier Research Institute, Suresnes, France
            Article
            25710 J Vasc Res 2000;37:26–31
            10.1159/000025710
            10720883
            4dcd2dd3-f671-4a04-af2e-5d88b41f8888
            © 2000 S. Karger AG, Basel

            Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher. Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug. Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

            History
            Page count
            Figures: 3, Tables: 1, References: 15, Pages: 6
            Categories
            Research Paper

            General medicine,Neurology,Cardiovascular Medicine,Internal medicine,Nephrology
            Video camera, high-speed,Videomicroscopy,Red blood cell velocity measurements,Microcirculation

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