Method for the study of the three‐dimensional orientation of the nuclei of myocardial cells in fetal human heart by means of confocal scanning laser microscopy

Usson, Yves; Parazza, F.; Jouk, Pierre‐Simon; Michalowicz, Gabrielle

doi:10.1111/j.1365-2818.1994.tb03454.x

Cited by 28 publications

(37 citation statements)

References 21 publications

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“…no trabeculae visible on the non-apical side) lateral invertion of the image preserved the standard orientation. The first of two techniques used for the evaluation of trabecular orientation used a polar histogram (adapted from Usson et al, 1994) of the lengths of the intertrabecular spaces oriented in each direction. The necessary data were obtained by measuring the length and orientation of individual intertrabecular spaces ( Fig.…”

Section: Methodsmentioning

confidence: 99%

“…either macroscopic (Streeter, 1979) or microscopic (Usson et al, 1994) examination of the ventricular The existence of a specific pattern of cardiomyocyte wall. During the embryonic period, the trabeculae are assembly in different regions of the heart is clear from radially arranged in the ventricles (Challice & pumping function (de Jong et al, 1992;Broekhuisen et al, 199.5;Hu & Keller, 1995).…”

Section: Introductionmentioning

confidence: 99%

“…These studies prompted us to develop a simple method for systematic, both qualitative and quantitative, description of the embryonic myoarchitecture. Several investigations have been performed on the myocardial architecture of human fetal and adult hearts (Usson et al, 1994;Sanchez-Quintana et al, 1995), and various methods have been developed for studying muscle fibre orientation (for review see Streeter, 1979). Jouk et al (1995) have recently provided a new computer-based method enabling threedimensional reconstruction and tracing of individual muscle fibres.…”

Section: Introductionmentioning

confidence: 99%

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A Quantitative Study of the Ventricular Myoarchitecture in the Stage 21?29 Chick Embryo Following Decreased Loading

Sedmera

Pexieder

et al. 1998

European Journal of Morphology

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During the early developmental period, ventricular myoarchitecture undergoes a transition from a smooth-walled cardiac tube, to left and right ventricular chambers filled with a sponge-like network of trabecular struts. We measured the quantitative changes of ventricular myocardium properties in normal stage 21-29 chick embryos and after chronic verapamil suffusion, which is known to decrease work load and decelerate ventricular growth. The morphologic parametres (compact layer thickness, ventricular wall composition, porosity of different layers and trabecular orientation) were determined from scanning electron micrographs of transversely dissected perfusionfixed hearts. A vascular bed of stage 21 chick embryos was suffused with 1 ng of verapamil at 1 pl per hour up to stages 24, 27 and 29 via a miniosmotic pump. From stage 24, the thickness of the compact myocardium in the left ventricle was greater than that of the right. The increase in thickness was minimal between stages 24 and 27, while the predominantly radially arranged trabeculae comprised up to 75% of total myocardial mass. The ratio of intertrabecular spaces to trabeculae (local porosity) decreased from the ventricular center (70%) towards the compact myocardium (0%). In verapamil-treated embryos, the hearts were smaller and showed delayed development. The compact myocardium was thinner than normal, and the proportion of trabeculae was higher than in controls. The local porosity values were similar in control and experimental groups. Decreased load resulted in delayed growth and morphogenesis, expressed as a persistence of trabeculae and a thinner compact myocardium. Embryonic heart pumping function is largely based on extensively developed trabeculation with regionally different properties.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A Quantitative Study of the Ventricular Myoarchitecture in the Stage 21?29 Chick Embryo Following Decreased Loading

Sedmera

Pexieder

et al. 1998

European Journal of Morphology

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“…Similar cells were examined in the present study, while they were maintained on glass slides. In this preparation, the nuclei maintain a particular figure and position during Zsectioning, thus allowing sorting of cells by the cell cycle phase using image cytometry, and three-dimensional localization of pericentromeres using CLSM (12,13,19,(36)(37)(38).…”

Section: Formation Of Pericentromeres and Entire Chromatinmentioning

confidence: 99%

“…Recent advances in biotechnology and the development of con-focal laser scanning microscopy (CLSM) have allowed a three-dimensional analysis of the nucleus, cell, or tissue structures (12,17,19,36,37,38). Using this technique, the morphology and distribution of chromosomes have been outlined in great detail in addition to identifying distinct chromosome territories (12,17).…”

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confidence: 99%

Differences in spatial localization and chromatin pattern during different phases of cell cycle between normal and cancer cells

et al. 1997

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We studied differences in chromatin patterns and the spatial localization of centromeres of chromosome 11 during the cell cycle between normal peripheral blood lymphocytes (PBL) and human promyclocytic leukemia cells (HL‐60) using fluorescence in situ hybridization. The pericentromeres in both cells were located at the periphery during Gq (quiescent) phase, but moved towards the nuclear center in G1 and mid‐S phase. During G2, the pericentromeres of PBL continued to move towards the nuclear center whereas those of HL‐60 returned to the periphery. The angle defining the spatial location of two pericentromeres, in reference to the center of the nucleus, increased in PBL cells from a mean of 67° during Gq phase to 106° during G1 phase (P < 0.01), and the two pericentromeres remained wide apart throughout the entire cell cycle. In HL‐60, the angle also increased during G1, but then decreased during mid‐S and G2 phases. Both cells exhibited pericentromeric signals during Gq that were round and compact, and the entire chromatin was loosely condensed. The signal became more loose and dispersed during the G1 and mid‐S phases. The pericentromere signal varied during G2 and was generally rod‐like or bipartite with condensation of the entire chromatin or chromosome‐like. Our results suggest that subtle but important differences in spatial localization of pericentromeres are present during the interphase between normal PBL and HL‐60 cells. Cytometry 27:327–335, 1997. © 1997 Wiley‐Liss, Inc.

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Three-Dimensional Confocal Laser Scanning Dna Ploidy Cytometry in Thick Histological Sections

et al. 1996

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DNA ploidy measurement by flow (FCM) or image cytometry (ICM) of single cell suspensions of solid tumour has prognostic value, but it would be a definite advantage if the assessment could be done on histological sections. However, this is usually not possible by means of standard ICM, due to the capping of nuclei in thin sections, or overlap in thick sections. Three‐dimensional (3D) microscopy by means of confocal laser scanning microscopy (CLSM) could solve this problem in theory but the results published so far are not very satisfactory. A new method has been developed in which the DNA content of haploid (human testis spermatozoa), diploid, tetraploid, octaploid (human and rat liver and human spermatogonia), and near‐triploid (human breast cancer) nuclei stained with YOYO‐1 iodide has been measured by a newly developed 3D image cytometry method (3DICM) in 20 μm thick histological sections. YOYO‐1 iodide is a new highly sensitive, specific, stoichiometric, and stable fluorescent dye for DNA. DNA ploidy of a breast cancer which was near‐triploid with FCM and ICM was also assessed with 3DICM in a tissue section adjacent to the section used for FCM and ICM and the results were compared. The integrated 3DICM fluorescence intensity showed good linearity (r=0·99) with the real DNA content of all nuclei analysed. In human tissue, the coefficient of variation of 3DICM for haploid (n=12), diploid (n=63), triploid (n=13), tetraploid (n=12), and octaploid (n=3) ploidy distributions was 5·1, 6·6, 4·2, 4·0, and 0·6 per cent, respectively (n=the number of nuclei). For the rat liver, the CV of the diploid (n=21), tetraploid (n=31), and octaploid (n=3) peaks was 6·7, 4·8, and 1·6 per cent, respectively. Repeated ‘blind’ measurements of nuclei with different DNA indices showed excellent reproducibility between different observers (r=0·98). It is concluded that the 3DICM method used is accurate, reproducible, and clinically feasible in thick histological sections. This is especially important in small lesions, or if the results of DNA ploidy measurement of single cell suspensions (by FCM) or imprints (by ICM) are inadequate.

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Method for the study of the three‐dimensional orientation of the nuclei of myocardial cells in fetal human heart by means of confocal scanning laser microscopy

Cited by 28 publications

References 21 publications

A Quantitative Study of the Ventricular Myoarchitecture in the Stage 21?29 Chick Embryo Following Decreased Loading

A Quantitative Study of the Ventricular Myoarchitecture in the Stage 21?29 Chick Embryo Following Decreased Loading

Differences in spatial localization and chromatin pattern during different phases of cell cycle between normal and cancer cells

Three-Dimensional Confocal Laser Scanning Dna Ploidy Cytometry in Thick Histological Sections

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