Imaging flow cytometry for automated detection of hypoxia‐induced erythrocyte shape change in sickle cell disease. Issue 6 (12th April 2014)

• Record Type: Journal Article
• Authors: van Beers, Eduard J.; Samsel, Leigh; Mendelsohn, Laurel; Saiyed, Rehan; Fertrin, Kleber Y.; Brantner, Christine A.; Daniels, Mathew P.; Nichols, James; McCoy, J. Philip; Kato, Gregory J.
• Is Part Of: American journal of hematology. Volume 89:Issue 6(2014:Jun.)
• Page Start: 598
• Page End: 603
• Publication Date: 2014-04-12
• Subjects: Hematology -- Periodicals; 616.15
• DOI: 10.1002/ajh.23699
• Languages: English
• ISSNs: 0361-8609

Abstract

<abstract abstract-type="main"> <title> <x xml:space="preserve">Abstract</x> </title> <p>In preclinical and early phase pharmacologic trials in sickle cell disease, the percentage of sickled erythrocytes after deoxygenation, an ex vivo functional sickling assay, has been used as a measure of a patient's disease outcome. We developed a new sickle imaging flow cytometry assay (SIFCA) and investigated its application. To perform the SIFCA, peripheral blood was diluted, deoxygenated (2% oxygen) for 2 hr, fixed, and analyzed using imaging flow cytometry. We developed a software algorithm that correctly classified investigator tagged "sickled" and "normal" erythrocyte morphology with a sensitivity of 100% and a specificity of 99.1%. The percentage of sickled cells as measured by SIFCA correlated strongly with the percentage of sickle cell anemia blood in experimentally admixed samples (<italic>R</italic> = 0.98, <italic>P</italic> ≤ 0.001), negatively with fetal hemoglobin (HbF) levels (<italic>R</italic> = −0.558, <italic>P</italic> = 0.027), negatively with pH (<italic>R</italic> = −0.688, <italic>P</italic> = 0.026), negatively with pretreatment with the antisickling agent, Aes‐103 (5‐hydroxymethyl‐2‐furfural) (<italic>R</italic> = −0.766, <italic>P</italic> = 0.002), and positively with the presence of long intracellular fibers as visualized by transmission electron microscopy (<italic>R</italic> = 0.799, <italic>P</italic> = 0.002). This study shows proof of principle that the automated, operator‐independent SIFCA is associated with predictable physiologic and clinical parameters and is altered by the putative antisickling agent, Aes‐103. SIFCA is a new method that may be useful in sickle cell drug development. Am. J. Hematol. 89:598–603, 2014. © 2014 Wiley Periodicals, Inc.</p> </abstract>