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Label-free quantitation of glycated hemoglobin in single red blood cells by transient absorption microscopy and phasor analysis.


ABSTRACT: As a stable and accurate biomarker, glycated hemoglobin (HbA1c) is clinically used to diagnose diabetes with a threshold of 6.5% among total hemoglobin (Hb). Current methods such as boronate affinity chromatography involve complex processing of large-volume blood samples. Moreover, these methods cannot measure HbA1c fraction at single-red blood cell (RBC) level, thus unable to separate the contribution from other factors such as RBC lifetime. Here, we demonstrate a spectroscopic transient absorption imaging approach that is able to differentiate HbA1c from Hb on the basis of their distinct excited-state dynamics. HbA1c fraction inside a single RBC is derived quantitatively through phasor analysis. HbA1c fraction distribution of diabetic blood is apparently different from that of healthy blood. A mathematical model is developed to derive the long-term blood glucose concentration. Our technology provides a unique way to study heme modification and to derive clinically important information void of bloodstream glucose fluctuation.

SUBMITTER: Dong PT 

PROVIDER: S-EPMC6510558 | biostudies-literature | 2019 May

REPOSITORIES: biostudies-literature

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Label-free quantitation of glycated hemoglobin in single red blood cells by transient absorption microscopy and phasor analysis.

Dong Pu-Ting PT   Lin Haonan H   Huang Kai-Chih KC   Cheng Ji-Xin JX  

Science advances 20190510 5


As a stable and accurate biomarker, glycated hemoglobin (HbA1c) is clinically used to diagnose diabetes with a threshold of 6.5% among total hemoglobin (Hb). Current methods such as boronate affinity chromatography involve complex processing of large-volume blood samples. Moreover, these methods cannot measure HbA1c fraction at single-red blood cell (RBC) level, thus unable to separate the contribution from other factors such as RBC lifetime. Here, we demonstrate a spectroscopic transient absorp  ...[more]

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