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Journal of Histochemistry and Cytochemistry, Vol. 45, 1097-1108, Copyright © 1997 by The Histochemical Society, Inc.

ARTICLE

Chromatin Condensation in Erythropoiesis Resolved by Multipixel Spectral Imaging: Differentiation Versus Apoptosis

Chana Rothmanna, Amos M. Cohenb, and Zvi Malika
a Life Sciences Department, Bar Ilan University, Ramat-Gan, Israel
b Hematology Unit, Rabin Medical Center, Golda Campus, Petach Tiqua, Israel

Correspondence to: Zvi Malik, Life Sciences Dept., Bar Ilan University, Ramat-Gan 52900, Israel.

Chromatin condensation and nuclear organization of May-Grunwald-Giemsa (MGG)-stained normal erythropoietic bone marrow cells and apoptotic red cell precursors were resolved by spectral bio-imaging. Multipixel spectra were obtained from single cells displaying a range of wavelengths of both transmitted and absorbed light. Two groups of spectra, of low- and high-intensity transmitted light, were revealed in the nuclei of each cell. The absorbance spectra served for the reconstruction of "absorbance images" depicting the affinity of MGG stain for the chromatin of proerythroblasts and of basophilic, polychromatic, and orthochromatic normoblasts. The localization of different spectral components in the nuclei was resolved employing two mathematical methods, spectral similarity mapping and principal component analysis. Novel structures of high symmetry revealing windmill-like organization were detected in basophilic, polychromatic, and orthochromatic normoblast cells. Matching structures were detected in apoptotic normoblasts obtained from an agnogenic myeloid metaplasia patient. Apoptosis was associated with a gradual breakdown of the ordered arrays in the nucleus. We propose that DNA cleavage may lead to fragmentation of the symmetrical windmill-like superstructure of the basic nuclear domains. (J Histochem Cytochem 45:1097-1108, 1997)

Key Words: multipixel spectroscopy, spectral similarity mapping, absorbance image, eigen images, apoptosis imaging, erythropoiesis imaging


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