Developments in optical experimental techniques have helped in elucidating how blood flows through microvessels. Although initial developments were encouraging, studies on the flow properties of blood in microcirculation have been limited by several technical factors, such as poor spatial resolution and difficulty obtaining quantitative detailed measurements at such small scales. Recent advances in computing, microscopy, and digital image processing techniques have made it possible to combine a particle tracking velocimetry (PTV) system with a confocal microscope. We document the development of a confocal micro-PTV measurement system for capturing the dynamic flow behavior of red blood cells (RBCs) in concentrated suspensions. Measurements were performed at several depths through 100-μm glass capillaries. The confocal micro-PTV system was able to detect both translational and rotational motions of individual RBCs flowing in concentrated suspensions. Our results provide evidence that RBCs in dilute suspensions (3% hematocrit) tended to follow approximately linear trajectories, whereas RBCs in concentrated suspensions (20% hematocrit) exhibited transversal displacements of about 2% from the original path. Direct and quantitative measurements indicated that the plasma layer appeared to enhance the fluctuations in RBC trajectories owing to decreased obstruction in transversal movements caused by other RBCs. Using optical sectioning and subsequent image contrast and resolution enhancement, the system provides previously unobtainable information on the motion of RBCs, including the trajectories of two or more RBCs interacting in the same focal plane and RBC dispersion coefficients in different focal planes.
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Abramoff, M., Magelhaes, P., Ram, S. 2004 Image Processing with Image J, Biophotonics Int. 11: 36-42.
Baker M., Wayland H. 1974 On-line volume flow rate and velocity profile measurement for blood in microvessels. Microvasc. Res. 7: 131-143.
Born G., Melling A., Whitelaw J. 1978 Laser Doppler microscope for blood velocity measurement. Biorheology 15: 163-172.
Caro, C., T. Pedley, R. Schroter, and W. Seed (1978). The Mechanics of the Circulation. Oxford: Oxford University Press
Chien, S. 1970 Shear dependence of effective cell volume as a determinant of blood viscosity. Science 168: 977-979.
Conchello J., Lichtman, J. 2005 Optical sectioning microscopy. Nat. Methods 2: 920–931.
Fahraeus, R., Lindqvist, T. 1931 The viscosity of the blood in narrow capillary tubes. Am. J. Physiol. 96: 562-568.
Fischer T., Stohr-Lissen M, Schmid-Schonbein, H. 1978 The red cell as a fluid droplet: tank tread-like motion of the human erythrocyte membrane in shear flow. Science 202: 894-896.
Fujiwara, H., Ishikawa T., Lima, R., Matsuki, N., Imai, Y., Kaji, H., Nishizawa, M., Yamaguchi, T. 2009 Red blood cell motions in high-hematocrit blood flowing through a stenosed microchannel. J. Biomech., 42: 838-843.
Gaehtgens P., Meiselman H., Wayland H. 1970 Velocity profiles of human blood at normal and reduced hematocrit in glass tubes up to 130 μ diameter. Microvasc. Res. 2: 13-23.
Goldsmith H. 1971 Red cell motions and wall interactions in tube flow, Fed. Proc. 30: 1578-1588.
Goldsmith H. 1971 Deformation of human red cells in tube flow, Biorheology 7: 235-242.
Goldsmith, H., Marlow, J. 1979 Flow behavior of erythrocytes. II. Particles motions in concentrated suspensions of ghost cells, J. Colloid Interface Sci. 71: 383-407.
Goldsmith, H., Turitto, V. 1986 Rheological aspects of thrombosis and haemostasis: basic principles and applications. ICTH-Report-Subcommittee on Rheology of the International Committee on Thrombosis and Haemostasis. Thromb. Haemost. 55: 415–435.
Inoue, S., and T. Inoue (2002) Direct-view high-speed confocal scanner: the CSU-10. In: Matsumoto B (ed.), Cell Biological Applications of Confocal Microscopy. San Diego: Academic Press, pp. 87–127.
Ishikawa, T. and Pedley, T. 2007 Diffusion of swimming model micro-organisms in a semi-dilute suspensions. J. Fluid Mech., 588: 437-462.
Kinoshita, H., Kaneda, S., Fujii, T.,Oshima, M. 2007 Three-dimensional measurement and visualization of internal flow of a moving droplet using confocal micro-PIV. Lab Chip 7: 338-346.
Lima, R. Analysis of the blood flow behavior through microchannels by a confocal micro-PIV/PTV system. Ph.D. Thesis, Tohoku University, Japan, 2007.
Lima, R., Ishikawa T., Imai, Y., Takeda, M., Wada, S., Yamaguchi, T. 2008 Radial dispersion of red blood cells in blood flowing through glass capillaries: role of hematocrit and geometry. J. Biomech. 41: 2188-2196.
Lima, R., Wada, S., Takeda, M., Tsubota, K., Yamaguchi, T. 2007 In vitro confocal micro-PIV measurements of blood flow in a square microchannel: the effect of the haematocrit on instantaneous velocity profiles. J. Biomech. 40: 2752-2757.
Lima, R., Wada, S., Tanaka, S., Takeda, M., Ishikawa, T., Tsubota, K., Imai, Y., Yamaguchi, T. 2008 In vitro blood flow in a rectangular PDMS microchannel: experimental observations using a confocal micro-PIV system. Biomed. Microdevices, 10: 153-167.
Lima, R., Wada, S., Tsubota, K.,Yamaguchi, T. 2006 Confocal micro-PIV measurements of three dimensional profiles of cell suspension flow in a square microchannel. Meas. Sci. Technol. 17: 797-808.
Meijering E., Smal I., Danuser G. 2006 Tracking in Molecular Bioimaging, IEEE Signal Process. Mag., 23: 46-53.
Meinhart C, Wereley S., Gray H. 2000 Volume illumination for two-dimensional particle image velocimetry. Meas. Sci. Technol. 11: 809-814.
Meinhart C, Wereley S, Santiago J. 1999 PIV measurements of a microchannel flow. Exp. Fluids 27: 414-419.
Miyazaki, H. and Yamaguchi, T. 2003 Formation and destruction of primary thrombi under the influence of blood flow and von Willebrand factor analysed by a D. E. M., Biorheology 40: 265-272.
Nash, G., Meiselman, H. 1983 Red cell and ghost viscoelasticity. Effects of hemoglobin concentration and in vivo aging. Biophys. J. 43: 63-73.
Park J, Choi C, and Kihm K 2004 Optically sliced micro-PIV using confocal laser scanning microscopy (CLSM). Exp. Fluids 37: 105-119.
Parthasarathi A., Japee S., Pittman R. 1999 Determination of red blood cell velocity by video shuttering and image analysis. Ann. Biomed. Eng. 27: 313-325.
Schmid-Schonbein, H., Wells, R. 1969 Fluid drop-like transition of erythrocytes under shear. Science 165: 288-291.
Shiga, T., Maeda N., Kon K. 1990 Erythrocyte rheology. Crit. Rev. Oncol. Hematol. 10: 9-48.
Sugii Y, Okuda R, Okamoto K, Madarame H 2005 Velocity measurement of both red blood cells and plasma of in vitro blood flow using high-speed micro PIV technique. Meas. Sci. Technol. 16: 1126-1130.
Tanaani T, Otsuki S, Tomosada N, Kosugi Y, Shimizu M., Ishida H. 2002 High-speed 1-frame/ms scanning confocal microscope with a microlens and Nipkow disks. Appl. Opt. 41: 4704-4708.
Tsubota, K., Wada, S., Yamaguchi, T. 2006 Particle method for computer simulation of red blood cell motion in blood flow. Comput. Methods Programs Biomed. 83: 139-146.
Uijttewaal W., Nijhof E., Heethaar R. 1994 Lateral migration of blood cells and microspheres in two-dimensional Poiseuille flow: a laser Doppler study. J. Biomech. 27: 35-42.
Vennemann P., K. Kiger, R. Lindken, B. Groenendijk, S. Stekelenburg-de Vos, T. Hagen, N. Ursem, R. Poelmann, J. Westerweel, B. Hierk 2006 In vivo micro particle image velocimetry measurements of blood-plasma in the embryonic avian heart. J. Biomech. 39: 1191-1200.
Wootton, D., Ku D. 1999 Fluid mechanics of vascular systems, diseases, and thrombosis. Annu. Rev. Biomed. Eng. 1: 299-329.
Yamaguchi, T., Ishikawa, T., Tsubota, K., Imai, Y., Nakamura M., Fukui T. 2006 Computational blood flow analysis—new trends and methods. J. Biomech. Sci. Eng. 1: 29-50.
This study was supported in part by the following grants: International Doctoral Program in Engineering from the Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT), “Revolutionary Simulation Software (RSS21)” next-generation IT program of MEXT; Grants-in-Aid for Scientific Research from MEXT and JSPS Scientific Research in Priority Areas (768) “Biomechanics at Micro- and Nanoscale Levels”, “Scientific Research (S) No. 19100008”.
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Lima, R., Ishikawa, T., Imai, Y. et al. Measurement of Individual Red Blood Cell Motions Under High Hematocrit Conditions Using a Confocal Micro-PTV System. Ann Biomed Eng 37, 1546–1559 (2009). https://doi.org/10.1007/s10439-009-9732-z
- Blood flow
- Red blood cells
- Confocal micro-PTV
- RBC dispersion