The applications of DNA cloning and fluorescent in situ hybridization (FISH) techniques have strengthened the hypothesis of an ordered chromatin structure in interphase nuclei, strongly suspected to vary with functional state. The nonrandom distribution of the centromeres and their dynamic rearrangement during the cell cycle have been well documented. A close proximity of specific centromeres to nucleoli has also been reported, but the functional meaning of this association is still unknown. In order to investigate whether the chromosome 1 centromere region to nucleolus association depends on the cell cycle and chromosome status, we combined FISH of probes specific for the lq12 region with Ki-67 nucleolar antigen fluorescent immunocytochemical (FICC) detection on the MCF-7 human breast cancer cell line and on the MRC-5 normal fibroblastic cell line. Both FISH and FICC signals were interactively localized in a one-step fluorescent microscopic observation and further analyzed using the Highly Optimized Microscope Environment (HOME) graphics microscope workstation, which provided computerized interactive marking of lq12 to nucleolus associations (lql2-nu) at the individual nucleus and nucleolus levels. This study confirms that centromeric regions, other than those adjacent to the major ribosomal cistrons, contribute to the perinucleolar chromatin and demonstrate that, during the cell cycle, the heterochromatic band lq12 is dynamically rearranged with regard to both the nuclear volume and the nucleoli. A relationship between the association of the chromosome 1 pericentromeric region with nucleoli and the nucleolar transcriptional activity is also strongly suggested. Key terms: Combination of FISH and immunocytochemistry, functional genome mapping, chromosome 1, nucleolus, interactive cytometry, HOME systemThe recent applications of DNA cloning and fluorescent in situ hybridization (FISH) techniques have strengthened the hypothesis of an ordered chromatin structure, strongly suspected to vary with functional state, in the interphase nuclei (2,(20)(21)(22)(23)27,29,32). Using specific antibodies or nucleic acid probes, several authors have demonstrated the nonrandom distribution and the cell cycle dependent rearrangement of centromeres in interphase nuclei (1,9,13,36,38). The proximity of specific chromosome centromeres and nucleoli has also been reported (3,14,16,23,24). The short distance between the secondary [nucleolus organizer regions (NORs)] and the primary (centromeres) constrictions in acrocentric chromosomes is most likely responsible for the centromere to nucleolus proximity, which is the case for chromosomes 13,14,15,21, and 22 bearing the ribosomal DNA in humans. However, this hypothesis does not apply when chromosomes such as 1 and 9 are involved (23,331 and when the number of chromosome to nucleolus associations is higher than the number of NOR-bearing chromosomes (25).The aim of this work was thus to investigate whether the chromosome 1 centromeric region to nucleolus association depends on the cel...
The Highly Optimized Microscope Environment (HOME) is a computerized microscope designed to assist pathologists and cytotechnicians in clinical routine tasks. The prototype system consists of a IBM-PC compatible computer and a light microscope in which a built-in highresolution computer display image is superimposed on the optical image of the specimen. Also, a manually operated encoding stage and objective turret encoder are used to provide continuous monitoring of the stage coordinates and microscope magnification to the computer. This allows any position on a slide to be uniquely defined and makes it possible to measure interactively lengths and areas larger than the size of the microscope field. Software, written in the C language and operating under the MS-DOSMS-Windows environment, is controlled by means of a mouse-driven cursor moving over menu light-buttons displayed on the microscope image. The HOME microscope workstation is potentially useful in a wide range of applications such as i) tagging information on particular cells and tissue structures that can thus be accurately located and relocated, ii) performing morphometric measurement, differential counting, and stereological assessment of biological specimens, and iii) training and educating laboratory personnel. Finally, HOME will offer in the near future a userfriendly interface for automatic image processing of cells and tissue entities in interactively selected specimen areas.Key terms: Microscopy, image cytometry, image histometry, computerized microscopy In modern hospitals, the pathology laboratory plays a n essential role in the diagnosis and prognosis of most serious diseases, and therefore in the subsequent medical decision-making and health care for the patient. In spite of this essential role, pathology practice has to date remained relatively untouched by recent developments in information technology: routine pathology is still carried out largely by microscopic examination of specimens by the pathologist, followed by the reporting of his opinion concerning the appearance of the specimen back to the requesting clinician. While this is satisfactory for the majority of specimens, for most types of pathological sample there is a "grey area" containing some specimens for which the result is not clear and is therefore subjective, unreliable, and poorly reproducible [lo].One approach to improving the diagnosis for such specimens is the introduction of quantitative analysis. Counting is the first quantitative approach which has been used extensively in haematology (differential white blood cell count), tumour pathology (mitotic index), and hormonal cytology (pycnotic index). Several other quantitative techniques for pathology have been investigated extensively for many years. Interactive computer morphometry, in which the user can delineate lengths and areas on a projected microscope image using a digital tablet or similar instrument, has been found to be diagnostically useful in many types of tumour pathology, bone pathology, and muscle patholo...
HOME is a new computerized microscope designed to assist pathologists and cytotechnicians in routine examinations. The HOME workstation is composed of a standard light microscope fitted with objective and stage encoders, and a built-in high resolution computer display which superimposes dialog, drawing, and messages onto the optical microscope image. The software runs under Windows 3.x and provides interactive facilities such as accurate localization and relocation of zones of interest, morphometric measurements, patient data access, and quality control processes.
In this paper, we present a new appronch to a computerized microscope to assist puthologists and cytotechnicians in routine exanzlnntions. The HOME workstutlon (Highly Optinzizeci Mici oscope Environnzent) i s conzyosed of a standui-ti light irzicroscoye .fitled with objective and stage encoders, iind N built-in high lasolution computer display which supei-rtiiyoses di-awrngs und messages onto the opticcil nzrci-oscope inzage. The so~flwure runs Windows 3.x und pi-ovrdes intermtrve facilities such as accurate localization and relocation of zone of interest, nzorphonzeti I C and densitometric nzeusurenzents, putient duta uccess unci quality conti-o I processes. THE IMPACT PROJECTThe diagnosis and prognosis of most severe diseases require microscopic examinations of slide-mounted specimens in order to evaluate the deviation of cell (cytology) and/or tissues (histology) from normal. In spite of the essential role of pathology in health care, this medical activity remains almost untouched by recent technological development : the routine clinical examinations are still cai-ried out by cytotechinicians or pathologists in a strictly manual mode and are qualitative, context-dependent and poorly reproducible. Quantitative microscopy (counting, stereology, morphometry and densitometry) is one approach to improving the quality of the diagnosis [ 11. The impact of this approach on pathology practice still remains limited because of low cost-effectiveness and poor user-friendliness.The IMPACT project is undertaken within the framework of the AIM (Advanced Informatics in Medicine) programme of the European Communities and involves several research laboratories, hospital pathology laboratories and industrial partners. The project specifically addresses the integration of computer teclmologies into the medical examinations performed at the microscope in cy t opa tho1 og y , 11 is t opa t h 0 1 og y ;in d cytogenetics hospital laboratories by designing and developing a microscope workstation based on a new concept named HOME (Highly Optimized Microscope Environment). HOME puts the emphasis on man-machine cornmunication and ergonomy for routine examinations in pathology. The HOME workstation has been designed to fulfil, given the constraints of the medical routine real world, the following requirements : -The user is more or less computer-innocent. Thus, the interactions between the U the computer should be le as possible and d by a unique means of namely, a mouse driven graphics interface. -The user should access a command environment split into four basic modules : (i) Data access : to perform the examination, the user needs access to differe information such as patient and specimen cl result of previous examinations, (ii) Measurements : accurate graphic tools should be available to mark and count relevant objects and to measure significant features (shape, area, length, densitometry, ratios), (iii) Control : the consistency between clinical data, current diagnosis and the adequacy of the operating features (size of the scanned...
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