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Imaging the Lateral Distribution of Fluorescently Labeled Membrane Components of Human Erythrocytes Under Deformation

Published online by Cambridge University Press:  02 July 2020

D.W. Knowles ,
N. Mohandas ,
C. Ortiz de Solorzano  and
S.J. Lockett
D.W. Knowles
Affiliation:
Life Sciences Division, Lawrence Berkeley National Laboratory, University of California, Berkeley, CA94720
N. Mohandas
Affiliation:
Life Sciences Division, Lawrence Berkeley National Laboratory, University of California, Berkeley, CA94720
C. Ortiz de Solorzano
Affiliation:
Life Sciences Division, Lawrence Berkeley National Laboratory, University of California, Berkeley, CA94720
S.J. Lockett
Affiliation:
Life Sciences Division, Lawrence Berkeley National Laboratory, University of California, Berkeley, CA94720
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Extract

The human erythrocyte membrane comprises many different biological macromolecules arranged into a cohesive two dimensional structure. It has long been a model system for studying cell membrane component interactions and their related underlying cell biology. Erythrocyte membrane rigidity emanates from its hexagonal semi-solid cytoskeletal network of spectrin polymers joined at junctional complexes by globular proteins. The network supports a 2D fluid double layer of lipid in which is dissolved a large array of receptor proteins some of which completely span the lipid bilayer and link to the underlying cytoskeletal network.

To study membrane component - component interactions, we have used the technique of fluorescence imaged microdeformation (1). This technique combines fluorescence labeling of specific membrane components, single cell microdeformation and state-of-the-art image collection and analysis to map the distribution of labeled components on the surface of the cell.

Type
Novel Approaches to Microscopy Of Living Cells
Information
Microscopy and Microanalysis , Volume 5 , Issue S2: Proceedings: Microscopy & Microanalysis '99, Microscopy Society of America 57th Annual Meeting, Microbeam Analysis Society 33rd Annual Meeting, Portland, Oregon August 1-5, 1999 , August 1999 , pp. 1044 - 1045
Copyright
Copyright © Microscopy Society of America

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References

1.Discher, D.E., Mohandas, N. & Evans, E.A.Molecular maps of red cell deformation: hidden elasticity and in situ connectivity. Science 266, 1032–1035 (1994)CrossRefGoogle ScholarPubMed
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3.Cho, M.R., Knowles, D.W., Smith, B.L, Moulds, J.J., Agre, P., Mohandas, N., Golan, D.E., Membrane dynamics of the water transport protein AQP1 in intact human red cells. Biophys. J. 76:1136–1144 (1999)CrossRefGoogle ScholarPubMed