title: Quantifying morphological alteration of RBC population from light scattering data.
abstract: Studies of RBC morphological alterations, despite their potential clinical and experimental application, are compromised due to lack of simple and rapid techniques. As a complementary approach toward quantitative microscopy, we have reconstituted morphological information from light scattering data obtained from flow cytometer. Normal and poikilocytic agent treated samples were analyzed by microscopy and respective morphological index (MI) was calculated from the morphology based scores assigned to RBC. The samples were simultaneously analyzed by flowcytometer and the scatter data were obtained. Accordingly, the best correlated parameters of both forward scatter and side scatter were chosen to formulate a suitable regression model with MI as response. Flow cytometry data was also verified with another instrument (BD FACS Verse) and the equation obtained was validated with separate set of samples. The multivariate regression analysis yields a quadratic model with MI as response (R2 = 0.96, p <  0.001). The flow cytometric data from both instruments were in good agreement (Intra class correlation ∼0.9, p <  0.001). The model was found to simulate the sample MI with high accuracy (R2 = 0.97, p <  0.001). This proposed method was verified to be simple, rapid, quantitative and cost effective for the measurement of morphological alteration of RBC.
authors.affiliation: Department of Physiology, University of Calcutta, Kolkata, India.|Department of Biochemistry, NRS Medical College, Kolkata, India.
authors.forename: Raghwendra
authors.initials: R
authors.lastname: Mishra
authors.affiliation: Department of Chemical Engineering, University of Calcutta, Kolkata, India.
authors.forename: Debasish
authors.initials: D
authors.lastname: Sarkar
authors.affiliation: Department of Physiology, University of Calcutta, Kolkata, India.
authors.forename: Sourav
authors.initials: S
authors.lastname: Bhattacharya
authors.affiliation: CU BD Centre of Excellence for Nanobiotechnology, University of Calcutta, Kolkata, India.|BD Biosciences, Salt Lake, Sector V, Kolkata, India.
authors.forename: Sanjaya
authors.initials: S
authors.lastname: Mallick
authors.affiliation: Department of Physiology, University of Calcutta, Kolkata, India.
authors.forename: Mousumi
authors.initials: M
authors.lastname: Chakraborty
authors.affiliation: Department of Physiology, University of Calcutta, Kolkata, India.|Department of Biochemistry, NRS Medical College, Kolkata, India.
authors.forename: Debarati
authors.initials: D
authors.lastname: Mukherjee
authors.affiliation: Department of Biochemistry, NRS Medical College, Kolkata, India.
authors.forename: Manoj
authors.initials: M
authors.lastname: Kar
authors.affiliation: Department of Physiology, University of Calcutta, Kolkata, India.
authors.forename: Roshnara
authors.initials: R
authors.lastname: Mishra
mesh_terms: D000328:Adult; D004912:Erythrocytes / Q000473:pathology*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D008027:Light; D008297:Male; D012542:Scattering, Radiation
keywords: Red blood cell; flow cytometry; light scattering; morphological alteration; morphological index