Segmentation of cervical cell images.

A major problem in the automation of cervical cytology screening is the segmentation of cell images. This paper describes various standard segmentation methods plus one which determines a segmentation threshold based on the stability of the perimeter of the cell as the threshold is varied. As well as contour, certain structural information is used to decide upon the threshold which separates cytoplasm from the background. Once the cytoplasm threshold is found, cytoplasm and nucleus are separated by simple clustering into three groups, cytoplasm, folded cytoplasm and nucleus. These techniques have been tested on 1500 cervical cells that belong to one of eight normal classes and five abnormal classes. A minimum Mahalanobis distance classifier was used to compare results. Manually thresholded cells were classified correctly 66.0% of the time for the 13 class problem and 95.2% of the time on the two (normal-abnormal) class problem. The contour tracing technique was 52.9% and 90.0% correct, respectively.     

Segmentation of microscopic cell scenes

Different methods for the automated segmentation of microscopic cell scenes are presented with examples. The techniques discussed include edge detection by thresholding, "blob" detection by split-and-merge algorithm, global thresholding using gray-level histograms, hierarchic thresholding using color information, global thresholding using two-dimensional histograms and segmentation by "blob" labeling. Methods are more robust against insignificant changes in the scene and perform more reliably as more a priori knowledge about the scene is incorporated in the segmentation algorithm. The inclusion of both photometric and geometric a priori knowledge can result in a high level of correct segmentations, the cost of which is increased computation time.     

Laser-based microscopic approaches: application to cell signaling in environmental lung disease.

Cell-imaging approaches using new laser-based technologies have a wide applicability to thefields of pathology and cell biology. Here, we present the application of several of these techniques, including confocal scanning laser microscopy (CSLM), laser scanning cytometry (LSC), and laser capture microdissection (LCM), to studies of cell signaling by environmental agents in lung disease. Using both cells in culture and lung tissue, we show that these technologies are powerful tools for understanding signal transduction cascades elicited by toxic agents, such as oxidants and asbestosfibers, and their relationship to the development of cell injury and proliferation, responses leading to lung disease and/or repair.     

Segmentation of microscopic images of small intestinal glands with directional 2-D filters

We present an image segmentation algorithm for small intestinal glands consisting of goblet cells that are evenly distributed and arranged in parallel at the base. Making use of the properties of the chain distribution of the goblet cells, directional 2-dimensional (2-D) linear filters with different orientations were designed to enhance the rims of the intestinal glands. Segmentations are based on the combined responses of the multiple zero-phase directional 2-D linear filters. For comparisons, outputs of combined directional filters are shown along with those of the comparable nondirectional Gaussian filters. Segmentation results of small intestinal glands of both normal and cancer cases are provided.     

Interpretation of scanning electron microscopic images of abraded forming bone surfaces

The experimental abrasion of forming bone surfaces was conducted so that such surfaces could be characterized. This is particularly important to bone remodeling studies utilizing scanning electron microscope (SEM) imaging of archeological material. Forming surfaces derived from subadult macaque cranial bone were treated by particle abrasion, water abrasion, sliding abrasion, brushing, manual rubbing, weight, exfoliation, chipping and replication. Acetic acid treatments were also performed. The effects of abrasive agents are specific but generally fall into rough (particle and water abrasion) and smooth (sliding abrasion, brushing, rubbing and weight) categories. Protohistoric human and Plio-Pleistocene hominid subadult craniofacial remains were observed with the SEM for comparison with experimental data. The more recent material appeared smooth, probably as a result of specimen preparation procedures using brushes. Surfaces were still interpretable as forming, however, using a more abrasion-resistant feature called intervascular ridging (IVR) described in this study. The IVR pattern is also recognized on the hominid sample, confirming the possibility of performing remodeling studies on abraded fossil material. The abrasion characteristics are somewhat more difficult to classify, however. Abrasion is defined and discussed relative to remodeling studies and taphonomy. The usefulness of the experimental data reported here, however, in paleoenvironmental reconstruction, has yet to be fully realized. Acid and mechanical preparation techniques are briefly addressed. It is concluded that it is possible to characterize a forming surface as abraded according to the findings of this study and that acid, if handled with care, will more likely preserve microanatomical surface detail. It would also be in everyone's interest to employ a less abrasive cleaning regime on archeological specimens.     

G-CSF: From granulopoietic stimulant to bone marrow stem cell mobilizing agent

G-CSF was among the first cytokines to be identified and rapidly transitioned into clinical medicine. Initially used to promote the production of neutrophils in patients with chemotherapy-induced neutropenia it helped to revolutionize the delivery of cancer therapy. Its ability to mobilize hematopoietic stem cells from the bone marrow into the blood was subsequently exploited, changing the face of hematopoietic stem cell transplantation. Today the knowledge gained in unraveling the mechanisms of stem cell mobilization by G-CSF is being explored as a means to increase chemosensitivity in hematological malignancies. This review provides a brief history of G-CSF and then focuses on recent advances in our understanding of G-CSF-induced stem cell mobilization and the potential clinical application of this knowledge in chemo-sensitization.     

Easily-handled method to isolate mesenchymal stem cells from coagulated human bone marrow samples

AIM: To establish an easily-handled method to isolate mesenchymal stem cells (MSCs) from coagulated human bone marrow samples.METHODS: Thrombin was added to aliquots of seven heparinized human bone marrow samples to mimic marrow coagulation. The clots were untreated, treated with urokinase or mechanically cut into pieces before culture for MSCs. The un-coagulated samples and the clots were also stored at 4 °C for 8 or 16 h before the treatment. The numbers of colony-forming unit-fibroblast (CFU-F) in the different samples were determined. The adherent cells from different groups were passaged and their surface profile was analyzed with flow cytometry. Their capacities of in vitro osteogenesis and adipogenesis were observed after the cells were exposed to specific inductive agents.RESULTS: The average CFU-F number of urokinase-treated samples (16.85 ± 11.77/10⁶) was comparable to that of un-coagulated control samples (20.22 ± 10.65/10⁶, P = 0.293), which was significantly higher than those of mechanically-cut clots (6.5 ± 5.32/10⁶, P < 0.01) and untreated clots (1.95 ± 1.86/10⁶, P < 0.01). The CFU-F numbers decreased after samples were stored, but those of control and urokinase-treated clots remained higher than the other two groups. Consistently, the numbers of the attached cells at passage 0 were higher in control and urokinase-treated clots than those of mechanically-cut clots and untreated clots. The attached cells were fibroblast-like in morphology and homogenously positive for CD44, CD73 and CD90, and negative for CD31 and CD45. Also, they could be induced to differentiate into osteoblasts and adipocytes in vitro.CONCLUSION: Urokinase pretreatment is an optimal strategy to isolate MSCs from human bone marrow samples that are poorly aspirated and clotted.     

Adipogenesis in a myeloid supporting bone marrow stromal cell line.

The bone marrow stroma contains pre-adipocyte cells which are part of the hemopoietic microenvironment. Cloned stromal cell lines differ both in their ability to support myeloid and lymphoid development and in their ability to undergo adipocyte differentiation in vitro. These processes have been examined in the +/+2.4 murine stromal cell line and compared to other stromal and pre-adipocyte cell lines. In long-term cultures, the +/+2.4 stromal cells support myeloid cell growth, consistent with their expression of macrophage-colony stimulating factor mRNA. However, despite the presence of mRNA for the lymphoid supportive cytokines interleukins 6 and 7, +/+2.4 cells failed to support stromal cell dependent B lineage lymphoid cells in vitro, suggesting that these stromal cells exhibit only a myelopoietic support function. The +/+2.4 cells differentiate into adipocytes spontaneously when cultured in 10% fetal bovine serum. The process of adipogenesis can be accelerated by a number of agonists based on morphologic and gene marker criteria. Following induction with hydrocortisone, methylisobutylxanthine, indomethacin, and insulin in combination, a time dependent increase in the steady state mRNA and enzyme activity levels of the following adipocyte specific genes was observed: adipocyte P2, adipsin, CAAT/enhancer binding protein, and lipoprotein lipase. In contrast, adipogenesis was accompanied by a slight decrease in the signal intensity of the macrophage-colony stimulating factor mRNA level, similar to that which has been reported in other bone marrow stromal cell lines. These data demonstrate that although the lympho-hematopoietic support function of pre-adipocyte bone marrow stromal cell lines is heterogeneous, they share a common mechanism of adipogenesis.     

Immunoelectron microscopic study of the insoluble antigens of bone marrow cells in rats

An immunoperoxidase study of the localization of insoluble antigens was carried out on the rat bone marrow cells. The effect of different fixatives and inhibitors of endogenous peroxidase on the cell ultrastructure and the preservation of immunoreactivity of the cell antigens. The best results were obtained while fixing with 1% paraformaldehyde and 0.05% glutaraldehyde mixture added with 0.5 saponin and using 10% acetic acid as an inhibitor of endogenous peroxidase. Differences were found in the localization of antigens and the intensity of immunoperoxidase staining in cells of different lines of differentiation and degree of maturity.     

Human bone marrow lymphocytes. III. Polyclonal activation of B lymphocytes in human bone marrow measured by a direct plaque-forming cell assay.

Lymphocytes from the bone marrow and peripheral blood of the same normal individuals were assayed simultaneously for blast transformation as well as polyclonal activation with differentiation to antibody-forming cells after stimulation with pokeweed mitogen. Blastogenic responses were measured by tritiated thymidine incorporation and antibody-forming cell assay. There was no significant difference between the blastogenic responses of lymphocytes in the peripheral blood compared to the bone marrow of the same individuals. However, differentiation to antibody-forming cells measured by the plaque-forming cell response was significantly greater in lymphocytes in the bone marrow as compared to peripheral blood of the same individuals. These studies demonstrate that the lymphocytes in human bone marrow are at a stage of differentiation whereby they can be readily induced to differentiation toward antibody production by polyclonal activation, even more so than peripheral blood lymphocytes. This supports the concept that the bone marrow is a major source of immunoglobulin production in man.