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Immunofluorescence Technique for Location Analysis of Artificial Red Blood Cells Cytoskeletal Proteins-Spectrin
Ying Wang1, Xiang Wang1*, Yao-Jin Li2, Feng Liu1, Ke Xiao1, Yan-Lian Xiong1
1Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University,Chongqing 400044, China; 2Southwest Hospital, Third Military Medical University, Chongqing 400038, China
Abstract: In this paper， we reported a new way to locate membrane protein of artificial red blood cells (RBC) and also provided a new method to estimate blood substitute by using immunofluorescence technique. By appling this method， we analyzed the artificial RBC reconstructed by our laboratory from the view of membrane structural proteins. The conformation of artificial RBC and its reconstrucing processes were analyzed by the comparison of distribution of spectrin in both natural RBC and artificial RBC， while spectrin αl was coloured in situ by indirect immunofluorescence method using fluorescence labeled rabbit-anti-human immunoglobulina and the fluorescence detected with laser scanning confocal microscope (LSCM) was excited by blue light of 488nm wavelength. It is similar to natural RBC， spectrin also distributed in the cytomembrane of RBC. Besides， our results further revealed the reconstructing processes of artificial RBC in vitro.
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