1. Total three-dimensional imaging of phase objects using defocusing microscopy: Application to red blood cells.
- Author
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Roma, P. M. S., Siman, L., Amara, F. T., Agero, U., and Mesquita, O. N.
- Subjects
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ERYTHROCYTES , *THREE-dimensional imaging in biology , *PHASE-contrast microscopy , *HYPERTONIC solutions , *REFRACTIVE index - Abstract
We introduce Defocusing Microscopy (DM), a bright-field optical microscopy technique able to perform total three-dimensional (3D) imaging of transparent objects. By total 3D imaging, we mean the determination of the actual shapes of the upper and lower surfaces of a phase object. We propose a methodology using DM and apply it to red blood cells subject to different osmolality conditions: hypotonic, isotonic, and hypertonic solutions. For each situation, the shapes of the upper and lower cell surface-membranes (lipid bilayer/cytoskeleton) are completely recovered, displaying the deformation of red blood cell (RBC) surfaces due to adhesion on the glass-substrate. The axial resolution of our technique allowed us to image surface-membranes separated by distances as small as 300 nm. Finally, we determine the volume, surface area, sphericity index, and RBC refractive index for each osmotic condition. [ABSTRACT FROM AUTHOR]
- Published
- 2014
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