A photometric method to analyze induced erythrocyte shape changes |
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| Institution: | 1. Department of Internal Medicine I, University Hospital of St. Poelten, Karl Landsteiner University of Health Sciences, Karl Landsteiner Institute for Nephrology and Hematooncology, St. Poelten, Austria;2. Division of Angiology, Medicine II, Medical University of Vienna, Vienna, Austria;1. Department of Pharmacy, General Hospital of Shenyang Military Area Command, Shenyang 110840, China;2. School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, Shenyang 110016, China;1. Nursing Research and Evidence-Based Practice, Houston Methodist Hospital, 6565 Fannin, MGJ 11-017, Houston, TX 77030, USA;2. UT Health School of Nursing, University of Texas Health Science Center at Houston, 6901 Bertner Street, Room 682, Houston, TX 77030, USA;1. Department of Pathology and Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114, USA;2. Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA;1. Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand;2. Research and Diagnostic Center for Emerging Infectious Diseases, Mekong Health Science Institue, Khon Kaen University, Khon Kaen, Thailand;3. Faculty of Medicine, Mahasarakram University, Maha Sarakram, Thailand;4. Centre of Malariology, Parasitology and Entomology, Ministry of Health, Vientiane, Lao Democratic People’s Republic;1. Department of Materials Science and Engineering, University of Delaware, 19716, USA;2. Department of Physics and Astronomy, University of Delaware, 19716, USA |
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| Abstract: | In this paper, a photometric method was introduced to quantify biochemically-induced red blood cell (RBC) shape changes when no shear force was acting on the cells. To obtain the photometric RBC shape parameter (RF1), a monolayer of point-attached RBCs was prepared on the floor of a flat flow chamber and the transmission of light perpendicular to the monolayer plane was measured: 1) in phosphate buffered saline with 0.1% bovine serum albumin (PBS+) and 2) in PBS+, containing a shape changing compound (in both, the RBCs were not deformed due to shear flow). To normalize the data, a third transmission value at a shear stress of 3 Pa was measured in PBS+ from the same RBC monolayer. To validate the photometric data, RF1 of RBCs exposed to shape changing agents was correlated by linear regression analysis with 1) data obtained with the tangent-counting technique (TC) and 2) the morphological index (MI). The coefficient of correlation was calculated at 0.95 for the TC data and 0.94 for the MI data, respectively. The sensitivity of the photometric method was tested with stomatocytogenic chlorpromazine (CP) and echinocytogenic sodium salicylate (SA). CP (2.5 μM) induced a significant decrease of RF1 to ?0.045 (N = 6 donors, p < 0.01), whereas SA (2.5 mM) increased RF1 to +0.027 significantly (N = 6, donors, p < 0.01). Both the CP-induced and the SA-induced shape changes appeared less than 2 min after application of the shape changing agents, and changed gradually within another 30 min when the agent was present in PBS+, partly disappearing within about 2 min after reincubation of the shape transformed RBCs in PBS+ not containing the agent. |
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