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Cited by 4 publications
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Fifty-three maturing bone marrow cells of the granulocyte cell series stained with Giemsa stain and magnified 1,000 times were scanned by a "computerized microscope" consisting of a LSI-11/23 microprocessor and a black-and-white video camera attached to a "frame grabber ." Each sampled cell was digitized into 70 X 70 pixels, each pixel representing 0.04 micron of the real image. The pixel gray values ranged between 0 and 255. Zero stood for white, 255 represented black, while the numbers in between stood for the various shades of gray. The cells represented six different stages of granulocytic maturation: myeloblast, promyelocyte, myelocyte, metamyelocyte , band form, and polymorphonuclear granulocyte. A discriminant analysis program selected 19 features best distinguishing between the six different cell types and computed five canonical discriminant functions defining a Space in which maturation was studied. In the Space, distance between two cells serves as a measure of similarity. The closer two cells are, the more similar they are and vice versa. This measure was applied here to express the degree of similarity between the neutrophil maturation classes, and since they represent states in the neutrophil life history, it is applicable also as a yardstick for the quantitation of differentiation. In the Space, the life history of a cell is represented by a trajectory originating in the myeloblast and terminating in the granulocyte state. Displacement along the trajectory represents cell maturation that is expressed relatively to the least differentiated state of the myeloblast. The further a cell from this state the more mature it is. The same yardstick also serves for differentiation rate estimates represented in the Space by displacement velocities that are derived from the known "transit times" of a cell in each state. The methodology is also applied for cell production estimates. Unlike other "computerized microscopes" serving for cell classification, the instrument described in this study is primarily a cell-comparator providing a precise measure of similarity between any two cells.
Fifty-three maturing bone marrow cells of the granulocyte cell series stained with Giemsa stain and magnified 1,000 times were scanned by a "computerized microscope" consisting of a LSI-11/23 microprocessor and a black-and-white video camera attached to a "frame grabber ." Each sampled cell was digitized into 70 X 70 pixels, each pixel representing 0.04 micron of the real image. The pixel gray values ranged between 0 and 255. Zero stood for white, 255 represented black, while the numbers in between stood for the various shades of gray. The cells represented six different stages of granulocytic maturation: myeloblast, promyelocyte, myelocyte, metamyelocyte , band form, and polymorphonuclear granulocyte. A discriminant analysis program selected 19 features best distinguishing between the six different cell types and computed five canonical discriminant functions defining a Space in which maturation was studied. In the Space, distance between two cells serves as a measure of similarity. The closer two cells are, the more similar they are and vice versa. This measure was applied here to express the degree of similarity between the neutrophil maturation classes, and since they represent states in the neutrophil life history, it is applicable also as a yardstick for the quantitation of differentiation. In the Space, the life history of a cell is represented by a trajectory originating in the myeloblast and terminating in the granulocyte state. Displacement along the trajectory represents cell maturation that is expressed relatively to the least differentiated state of the myeloblast. The further a cell from this state the more mature it is. The same yardstick also serves for differentiation rate estimates represented in the Space by displacement velocities that are derived from the known "transit times" of a cell in each state. The methodology is also applied for cell production estimates. Unlike other "computerized microscopes" serving for cell classification, the instrument described in this study is primarily a cell-comparator providing a precise measure of similarity between any two cells.
Erythroid cell types were quantitatively assessed on the basis of the analysis of their nuclear images after Feulgen and Heidenhain blue counterstaining. An original nonsupervised classification method, developed in our laboratory, was applied to cell cycle analysis. Results obtained suggest that, in the four proliferative generation cycles, which concur with the successive morphological stages, there is a sequential flow of cells from one morphological stage to the next by cell cycling. We show that the chromatin pattern alone made it possible to set up a metric of the whole differentiation process occurring in the normal human erythroblastic lineage in which proliferating events occur. Key terms: Cell cycle analysis, differentiation in erythroblastic lineage, erythropoiesisErythropoiesis is an appropriate model for analyzing descriptors obtainable by means of high-resolution imcell cycle kinetic events and the maturation process in age analysis may become more accurate and convenient morphological changes during the evolution from prim-for functional interpretation (36,43,44). itive precursor cell to the mature erythrocyte, Differen-In a previous paper (261, we described a quantitative tiation of erythroid cells proceeds through a series of color image analysis of the normal maturation sequence recognizable morphological changes, which give rise to of the erythroblastic May-Grunwald-Giemsa (MGG) a cell type classification used by hematologists. The stained cells using the SAMBA 200 image analyzer. We subsequent stages of erythroid maturation are termed concluded that the machine successfully reproduced the mature erythrocyte is continuous although arbitrarily "he structural changes occurring during differentia-broken up by hematologists. tion are associated with striking biochemical alterationsThe present investigation deals with a combined quanas shown by combined methods of morphology and au-titative study of cell proliferation and cell differentiation toradiography (10,16,18,21,30,33,35), Unfortunately, of Feulgen Heidenhaim CFH1 blue-stained erythroid cells, these methods are time-consuming, and discrepancies in based on the concept that the functional properties of a the results reported were probably due to errors in the cell (DNA synthesis and template DNA activity) are experimental techniques used (34). A new approach to reflected in chromatin pattern. The nonsupervised this problem has been provided by analytical cytology method of classification (3,4) has the advantage of permethods based on computer-assisted cell-image process-mitting cell kinetic analysis from a single sample of a ing. The results already obtained by Zajicek et al. (46,471 cell population and does not require any a priori knowland Dormer et al. (17,19,20) for the characterization of edge about the proliferating state of the cells or a kinetic the grey-level images of erythroblasts at the various model of cell population. The results obtained will be steps of the maturation process have shown that it is discussed in terms of the coo...
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