# Translation of the Morphological Hallmarks of Dyserythropoiesis to Objective Morphometric Parameters by Imaging Flow Cytometry

## TL;DR

This study shows how imaging flow cytometry can objectively detect abnormal red blood cell development in myelodysplastic neoplasms.

## Contribution

The study introduces novel objective morphometric parameters for dyserythropoiesis using imaging flow cytometry.

## Key findings

- MDS cells showed increased cytoplasmic contrast and variance compared to normal bone marrow.
- Abnormal nuclei and binucleated erythroblasts were significantly more common in MDS samples.
- MDS erythroblasts had decreased nuclear condensation and increased cell size across all maturation stages.

## Abstract

Imaging flow cytometry (IFC) is a unique method combining multiparameter flow cytometry (MFC) with morphological evaluation of single cells. Since both analyses are integrated in the diagnostic work‐up of myelodysplastic neoplasms (MDS), we wanted to explore the possibilities of IFC as a diagnostic tool for MDS, with focus on dyserythropoiesis.

We analysed fresh bone marrow (BM) aspirates from 26 patients with untreated MDS and MDS/MPN and compared them with 12 normal BM specimens (NBM) exploring the cytoplasmic compartment, nuclear abnormalities, and megaloblastoid changes.

The cytoplasmic compartment in MDS showed higher contrast and variance values compared to NBM (p < 0.001). Cells with abnormal nuclei and binucleated forms were significantly increased in MDS compared to NBM (p < 0.05 for both features). Most binucleated forms were found in the mature compartment, and many of them were G1 phase arrested. All maturation stages showed a significant increase in cell size in MDS compared to NBM (p < 0.001). In addition, we found decreased nuclear condensation combined with increased cell size for all erythropoietic maturation stages in MDS compared to NBM (p < 0.001). Finally, our previously described MDS‐specific aberrant population of mature erythroblasts with decreased expression of CD36 and/or CD71 showed denser chromatin than both the mature erythropoietic MDS cells without immunophenotypic aberrancies (p < 0.001) and NBM (p = 0.024).

IFC can detect the major morphological changes associated with dyserythropoiesis in MDS, including the novel findings of increased cytoplasmic texture and bilobated non‐proliferating erythroblasts, allowing for objectivity and standardization.