Abstract
The photophysical properties of human sickle cell disease (SCD) Hemoglobin (Hb) is characterized by multi-photon microscopy (MPM). The intrinsic two-photon excited fluorescence (TPEF) signal associated with extracted hemoglobin was investigated and the solidified SCD variant (HbS) was found to demonstrate broad emission peaking around 510 nm when excited at 800 nm. MPM is used to dynamically induce and image HbS gelling by photolysis of deoxygenated HbS. For comparison, photolysis conditions were applied to a healthy variant of human hemoglobin (HbA) and found to remain in solution not forming fibers. The use of this signal to study the mechanism of HbS polymerization associated with the sickling of SCD erythrocytes is discussed.
Highlights
Sickle cell disease (SCD), the most common hereditary blood disorder worldwide, causes a number of health complications including chronic pain, anemia, chronic infection and stroke
It has been thought for decades that the polymerization of HbS into rigid fibers is the primary pathogenic event causing the deformation of red blood cells [1,2,3]
To compliment microscopic investigations of SCD hemoglobin, multi-photon microscopy (MPM) is proposed as a means of direct visualization of the pathogenic sickling event by utilizing the intrinsic two-photon excited fluorescence (TPEF) signature described here
Summary
Sickle cell disease (SCD), the most common hereditary blood disorder worldwide, causes a number of health complications including chronic pain, anemia, chronic infection and stroke. It is caused by a single point mutation resulting in an amino acid substitution in the oxygen transport molecule hemoglobin (Hb). Cytological and rheological methods have been demonstrated to study macroscopic effects of various environmental and treatment conditions on the sickling rate [9, 11] in addition to older, direct microscopic visualization utilized to study the more fundamental, sub-cellular kinetics of the sickling mechanism [4, 12]. The use of MPM as an investigative research tool to further illuminate environmental conditions impacting the dynamics of HbS polymerization is discussed
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