Morphometric and colorimetric analysis of peripheral blood smears lymphocytes in B-cell disorders: Proposal for a scoring system

Distinguishing leukemic phases of B-cell disorders in peripheral blood smea rs is well recognized to be difficult in some cases since it depends on sub tle and subjective criteria. In order to quantify cytological features and to assess objective descriptions, a morphometric analysis was performed on 83 peripheral blood smears of B-cells disorders (n=77) and healthy donors ( n=6). Using standardized May-Grunwald Giemsa staining, standardized image a cquisition system and well defined microscopic fields, we have analyzed lym phoid cells, measuring morphometric and color parameters. By combining seve n relevant morphometric criteria (the nuclear shape, the cellular shape and area, the nucleo-cytoplasmic ratio, the nuclear red/blue ratio, the cytopl asmic green/blue ratio and the proportion of cells with nucleolus), we have established a score that could range from a minimum of -3 (large B-CLL typ e) to a maximum of +8 (large MCL type): negative scores corresponds to diff erent types of B-CLL (n=30), including "atypical B-CLL" (n=6), the score ze ro correspond to healthy donors (n=6) used as baseline, the positive score values correspond to +1 for Follicular lymphoma (n=2), +3 for Splenic Lymph oma with Villous Lymphocytes (n=12), +4 for Hairy Cell Leukemia (n=7), +5 f or Hairy Cell Leukemia-variant (n=2), +6 for B-prolymphocytic leukemia (n=6 ) and +7 and +8 for most Mantle Cell Lymphoma (n=18). Testing T-cell disord ers samples (n=10) using the same protocol, the profile is different and ca nnot be confused with B-cell diseases. Our scoring system indicates that measurement of some common morphologic fe atures in standardized conditions provides objective criteria to characteri ze those diseases and might be helpful for diagnosis.