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Proceedings Paper

Developing strategies to enhance loading efficiency of erythrosensors
Author(s): Sandra C. Bustamante Lopez; Sarah C. Ritter; Kenith E. Meissner
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Paper Abstract

For diabetics, continuous glucose monitoring and the resulting tighter control of glucose levels ameliorate serious complications from hypoglycemia and hyperglycemia. Diabetics measure their blood glucose levels multiple times a day by finger pricks, or use implantable monitoring devices. Still, glucose and other analytes in the blood fluctuate throughout the day and the current monitoring methods are invasive, immunogenic, and/or present biodegradation problems. Using carrier erythrocytes loaded with a fluorescent sensor, we seek to develop a biodegradable, efficient, and potentially cost effective method to continuously sense blood analytes. We aim to reintroduce sensor-loaded erythrocytes to the bloodstream and conserve the erythrocytes lifetime of 120 days in the circulatory system. Here, we compare the efficiency of two loading techniques: hypotonic dilution and electroporation. Hypotonic dilution employs hypotonic buffer to create transient pores in the erythrocyte membrane, allowing dye entrance and a hypertonic buffer to restore tonicity. Electroporation relies on controlled electrical pulses that results in reversible pores formation to allow cargo entrance, follow by incubation at 37°C to reseal. As part of the cellular characterization of loaded erythrocytes, we focus on cell size, shape, and hemoglobin content. Cell recovery, loading efficiency and cargo release measurements render optimal loading conditions. The detected fluorescent signal from sensor-loaded erythrocytes can be translated into a direct measurement of analyte levels in the blood stream. The development of a suitable protocol to engineer carrier erythrocytes has profound and lasting implications in the erythrosensor’s lifespan and sensing capabilities.

Paper Details

Date Published: 28 February 2014
PDF: 6 pages
Proc. SPIE 8951, Optical Diagnostics and Sensing XIV: Toward Point-of-Care Diagnostics, 895114 (28 February 2014); doi: 10.1117/12.2042119
Show Author Affiliations
Sandra C. Bustamante Lopez, Texas A&M Univ. (United States)
Sarah C. Ritter, Texas A&M Univ. (United States)
Kenith E. Meissner, Texas A&M Univ. (United States)


Published in SPIE Proceedings Vol. 8951:
Optical Diagnostics and Sensing XIV: Toward Point-of-Care Diagnostics
Gerard L. Coté, Editor(s)

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