The Microrheology of Sickle Hemoglobin Gels

• Published in: Biophysical journal Vol. 99; no. 4; pp. 1149 - 1156
• Main Authors: Zakharov, Mikhail N., Aprelev, Alexey, Turner, Matthew S., Ferrone, Frank A.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 09.08.2010; Biophysical Society; The Biophysical Society
• Subjects: Biophysics; Carboxyhemoglobin - metabolism; Droplets; Elastic Modulus; Emulsions; Gels; Hemoglobin; Hemoglobin, Sickle - metabolism; Hemorheology - physiology; Humans; Mathematical models; Modulus of elasticity; Pathology; Polymerization; Protein; Rheological properties; Rheology; Sickle cell disease; Temperature
• ISSN: 0006-3495; 1542-0086
• DOI: 10.1016/j.bpj.2010.04.079

Sickle cell disease is a rheological disease, yet no quantitative rheological data exist on microscopic samples at physiological concentrations. We have developed a novel method for measuring the microrheology of sickle hemoglobin gels, based on magnetically driven compression of 5- to 8-μm-thick emulsions containing hemoglobin droplets ∼80 μm in diameter. Using our method, by observing the expansion of the droplet area as the emulsion is compressed, we were able to resolve changes in thickness of a few nanometers with temporal resolution of milliseconds. Gels were formed at various initial concentrations and temperatures and with different internal domain structure. All behaved as Hookean springs with Young's modulus from 300 to 1500 kPa for gels with polymerized hemoglobin concentration from 6 g/dl to 12 g/dl. For uniform, multidomain gels, Young's modulus mainly depended on the terminal concentration of the gel rather than the conditions of formation. A simple model reproduced the quadratic dependence of the Young's modulus on the concentration of polymerized hemoglobin. Partially desaturated samples also displayed quadratic concentration dependence but with a smaller proportionality coefficient, as did samples that were desaturated in steps; such samples were significantly less rigid than gels formed all at once. The magnitude of the Young's modulus provides quantitative support for the dominant models of sickle pathophysiology.