scholarly journals The fusion of trout spermatozoa with Chinese hamster fibroblasts

1982 ◽  
Vol 53 (1) ◽  
pp. 307-321
Author(s):  
J.H. Yip ◽  
N.C. Bols
Keyword(s):  
Electron Microscopy ◽  
Polyethylene Glycol ◽  
Light Microscopy ◽  
Chinese Hamster ◽  
Cell Types ◽  
Culture Conditions ◽  
Peg Treatment ◽  
Sperm Nuclei

Polyethylene glycol (PEG) was used to fuse trout sperm with Chinese hamster fibroblasts (CHW-1102). As judged by light microscopy, PEG significantly increased the number of associations between the two cell types. Electron microscopy revealed that the sperm nuclei were in the cytoplasm of CHW-1102 in cultures that had been treated with PEG and in phagosomes of CHW-1102 in cultures not treated with PEG. In the cytoplasm many sperm nuclei had an extensive network of fibres whereas before fusion the sperm nuclei contained chromatin blocks which were packed together tightly. In phagosomes they consisted of dispersed blocks of chromatin. The chromatin of the few sperm that were found outside CHW-1100 cells but in the same culture that was treated with PEG also lacked fibres and consisted of loosely packed blocks. Most nuclei were unaltered by PEG treatment alone although in a few the packing of the chromatin blocks was loosened. Thus the decondensation of sperm nuclei within CHW-1102 cells appears to be brought about by the mammalian cytoplasm rather than by digestion in phagosomes, the culture conditions, or the treatment used to induce fusion.

THE FUSION OF ERYTHROCYTES BY TREATMENT WITH PROTEOLYTIC ENZYMES AND POLYETHYLENE GLYCOL

1976 ◽  
Vol 18 (3) ◽  
pp. 503-512 ◽  
Author(s):  
James X. Hartmann ◽  
J. D. Galla ◽  
D. A. Emma ◽  
K. N. Kao ◽  
O. L. Gamborg
Keyword(s):  
Electron Microscopy ◽  
Molecular Weight ◽  
Polyethylene Glycol ◽  
Proteolytic Enzymes ◽  
Fusion Process ◽  
Significant Extent ◽  
Multinucleated Cells ◽  
Peg Treatment ◽  
Previously Treated ◽  
High Degree

Polyethylene glycol (PEG) has been utilized to induce homokaryocyte formation in avian and mammalian erythrocytes previously treated with proteolytic enzymes. PEG of molecular weight 6,000–7,500 was found superior to 1,500 and 20,000 MW PEG. Cells exposed to protease alone, prior to PEG treatment, fused to a high degree (60–95% multinucleated cells), whereas trypsin or pepsin treatment alone allowed very little fusion (2.5%). Trypsin lowered the effectiveness of protease when used in combination. Cells which were not treated with proteolytic enzymes agglutinated in the presence of PEG but did not fuse to a significant extent (0.01%). Fusion was also markedly dependent upon the rate at which PEG was eluted during the fusion process. Electron microscopy indicated that fusion began during the elution of PEG from the agglutinated cells.

scholarly journals Analysis by Light, Scanning, and Transmission Microscopy of the Intima Synovial of the Temporomandibular Joint of Human Fetuses during the Development

2014 ◽  
Vol 2014 ◽  
pp. 1-6
Author(s):  
Carlos Sabu Alvez ◽  
Luis Otavio Carvalho de Moraes ◽  
Sergio R. Marques ◽  
Roberto C. Tedesco ◽  
Leandro J. C. Harb ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Light Microscopy ◽  
Temporomandibular Joint ◽  
Cell Types ◽  
Human Fetuses ◽  
Transmission Microscopy ◽  
Joint Cavity ◽  
Transmission Electron ◽  
Intima Layer

Objective. To characterize morphologically and ultrastructurally using light microscopy, the scanning electron microscopy and transmission electron microscopy the intima synovial of the temporomandibular joint (TMJ) of human fetuses between the 10th and the 38th week of development. Materials and Methods. The TMJ was dissected bilaterally in 37 human fetuses belonging to the Institute of Embryology of the University Complutense of Madrid and of the Federal University of São Paulo. Results. The outcome by light microscopy showed the morphology of the TMJ and that the formation of inferior joint cavity precedes the superior joint cavity and the presence of blood vessels in the synovial. Conclusion. By scanning and transmission electron microscopy we observed the presence of two well-defined cell types in the intima layer of synovial of the TMJ of human fetuses, macrophage-like type A cell and fibroblast-like type B cell, and the presence of the a third cell type, defined by the name of intermediate lining cell in the intima layer of the synovial.

The Relationship Between Pit Membrane Ultrastructure and Chemical Impregnability of Wood

Holzforschung ◽  
1999 ◽  
Vol 53 (4) ◽  
pp. 341-346 ◽  
Author(s):  
Adya Singh ◽  
Bernard Dawson ◽  
Robert Franich ◽  
Faye Cowan ◽  
Jeremy Warnes
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Light Microscopy ◽  
Cell Types ◽  
Transmission Electron ◽  
Pit Membrane ◽  
Before And After ◽  
Ray Cells ◽  
Membrane Ultrastructure ◽  
The Relationship

Summary The woods of Alder and Eucalypt were examined by light microscopy before and after a chemical treatment by the Indurite process to increase the hardness of the wood. The pattern of wood cell impregnation for Alder differed significantly from Eucalypt in some respects. In Alder wood all cell types eg. vessels, fibres and rays, were impregnated in similar proportions. In comparison, in Eucalypt wood the impregnation material was largely confined to ray cells and the lumina of vessels; other cell types were either not impregnated or impregnated in very small numbers. Transmission electron microscopy of Alder and Eucalypt woods suggests that ultrastructural differences in the texture and porosity of pit membranes may be the main reason for the observed differences between these wood species with regard to their impregnability by the impregnation material used.

In Vitro Evaluation of a 3D PLGA–TCP Composite Scaffold in an Experimental Bioreactor

2009 ◽  
Vol 24 (1_suppl) ◽  
pp. 75-83 ◽  
Author(s):  
Antti A. Mäkitie ◽  
Yongnian Yan ◽  
Xiaohong Wang ◽  
Zhuo Xiong ◽  
Kaija-Stiina Paloheimo ◽  
...  
Keyword(s):  
Composite Scaffold ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Cell Types ◽  
Human Cell Line ◽  
Culture Conditions ◽  
Poly Lactic Acid ◽  
Scaffold Material ◽  
Cultivation In Vitro

A 3D poly(lactic acid-co-glycolic acid)/tricalcium phosphate (PLGA-TCP) composite scaffold, generated with the low-temperature deposition modeling rapid prototyping technique, was tested for its viability in a 3D cell cultivation in vitro. The aim was to find optimal cell culture conditions for the selected scaffold material and to monitor cell division, differentiation, and migration of selected cell types in this environment. In addition, the behavior and cell-matrix interactions of selected cell types were monitored as well as the biodegradation rate of the tested scaffold material. Chinese hamster ovary cells as well as a human cell line 293 epithelial cells were cultured on the scaffolds. A variety of different preconditioning protocols were deployed to prepare the scaffolds before seeding with the cells. Cell cultivations were conducted for 1–4 weeks and the coverage of the luminal surfaces was analyzed with light microscopy. Long cultivation periods were required to achieve partial coverage of the luminal surfaces of the scaffolds. Tissue engineering with 3D cell cultures and biomaterials represents a promising approach for organ manufacturing research. It may have potential for eventual on-demand high-throughput production of artificial tissues but the process has many challenges. The culture system in a well controlled bioreactor environment is discussed.

Two- or three-way observations of the identical cell elements within the same sections by LM, TEM and/or SEM

1978 ◽  
Vol 36 (2) ◽  
pp. 82-83 ◽  
Author(s):  
Nakazo Watari ◽  
Yasuaki Hotta ◽  
Yoshio Mabuchi
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Fine Structure ◽  
Light Microscopy ◽  
Thin Sections ◽  
Transmission Electron ◽  
Identical Cell ◽  
Electron Microscopes ◽  
Scanning Electron

It is very useful if we can observe the identical cell elements within the same sections by light microscopy (LM), transmission electron microscopy (TEM) and/or scanning electron microscopy (SEM) sequentially, because, the cell fine structure can not be indicated by LM, while the color is; on the other hand, the cell fine structure can be very easily observed by EM, although its color properties may not. However, there is one problem in that LM requires thick sections of over 1 μm, while EM needs very thin sections of under 100 nm. Recently, we have developed a new method to observe the same cell elements within the same plastic sections using both light and transmission (conventional or high-voltage) electron microscopes.In this paper, we have developed two new observation methods for the identical cell elements within the same sections, both plastic-embedded and paraffin-embedded, using light microscopy, transmission electron microscopy and/or scanning electron microscopy (Fig. 1).

Study of the Molecular Architecture of Keratin Filaments by Electron Microscopy

1982 ◽  
Vol 40 ◽  
pp. 118-119
Author(s):  
U. Aebi ◽  
P. Rew ◽  
T.-T. Sun
Keyword(s):  
Electron Microscopy ◽  
Structural Organization ◽  
Cell Types ◽  
Structural Features ◽  
Molecular Architecture ◽  
The Past ◽  
Keratin Filaments ◽  
Different Types ◽  
10 Nm Filaments ◽  
Different Cell Types

Various types of intermediate-sized (10-nm) filaments have been found and described in many different cell types during the past few years. Despite the differences in the chemical composition among the different types of filaments, they all yield common structural features: they are usually up to several microns long and have a diameter of 7 to 10 nm; there is evidence that they are made of several 2 to 3.5 nm wide protofilaments which are helically wound around each other; the secondary structure of the polypeptides constituting the filaments is rich in ∞-helix. However a detailed description of their structural organization is lacking to date.

Tumor Diagnosis

1982 ◽  
Vol 40 ◽  
pp. 262-265
Author(s):  
Bruce Mackay
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Differential Diagnosis ◽  
Light Microscopy ◽  
Clinical Examination ◽  
Ultrastructural Study ◽  
Diagnostic Procedures ◽  
Specific Diagnosis ◽  
Transmission Electron ◽  
Microscopic Diagnosis

The broadest application of transmission electron microscopy (EM) in diagnostic medicine is the identification of tumors that cannot be classified by routine light microscopy. EM is useful in the evaluation of approximately 10% of human neoplasms, but the extent of its contribution varies considerably. It may provide a specific diagnosis that can not be reached by other means, but in contrast, the information obtained from ultrastructural study of some 10% of tumors does not significantly add to that available from light microscopy. Most cases fall somewhere between these two extremes: EM may correct a light microscopic diagnosis, or serve to narrow a differential diagnosis by excluding some of the possibilities considered by light microscopy. It is particularly important to correlate the EM findings with data from light microscopy, clinical examination, and other diagnostic procedures.

Immunocytochemistry. A Review of Methodology for Electron Microscopic Applicaiton

1974 ◽  
Vol 32 ◽  
pp. 328-329
Author(s):  
Joseph E. Mazurkiewicz
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Ultrastructural Level ◽  
Electron Microscopic ◽  
Biological Interest ◽  
Investigative Approach ◽  
Immunologic Activity ◽  
Dense Label

Immunocytochemistry is a powerful investigative approach in which one of the most exacting examples of specificity, that of the reaction of an antibody with its antigen, isused to localize tissue and cell specific molecules in situ. Following the introduction of fluorescent labeled antibodies in T950, a large number of molecules of biological interest had been studied with light microscopy, especially antigens involved in the pathogenesis of some diseases. However, with advances in electron microscopy, newer methods were needed which could reveal these reactions at the ultrastructural level. An electron dense label that could be coupled to an antibody without the loss of immunologic activity was desired.

Electron Microscopy of Mycoplasma Pneumoniae

1971 ◽  
Vol 29 ◽  
pp. 258-259 ◽  
Author(s):  
E. S. Boatman ◽  
G. E. Kenny
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Growth Phase ◽  
Hela Cells ◽  
Sulfonic Acid ◽  
Gamma Globulin ◽  
Horse Serum ◽  
Morphological Aspects ◽  
The Family

Information concerning the morphology and replication of organism of the family Mycoplasmataceae remains, despite over 70 years of study, highly controversial. Due to their small size observations by light microscopy have not been rewarding. Furthermore, not only are these organisms extremely pleomorphic but their morphology also changes according to growth phase. This study deals with the morphological aspects of M. pneumoniae strain 3546 in relation to growth, interaction with HeLa cells and possible mechanisms of replication.The organisms were grown aerobically at 37°C in a soy peptone yeast dialysate medium supplemented with 12% gamma-globulin free horse serum. The medium was buffered at pH 7.3 with TES [N-tris (hyroxymethyl) methyl-2-aminoethane sulfonic acid] at 10mM concentration. The inoculum, an actively growing culture, was filtered through a 0.5 μm polycarbonate “nuclepore” filter to prevent transfer of all but the smallest aggregates. Growth was assessed at specific periods by colony counts and 800 ml samples of organisms were fixed in situ with 2.5% glutaraldehyde for 3 hrs. at 4°C. Washed cells for sectioning were post-fixed in 0.8% OSO4 in veronal-acetate buffer pH 6.1 for 1 hr. at 21°C. HeLa cells were infected with a filtered inoculum of M. pneumoniae and incubated for 9 days in Leighton tubes with coverslips. The cells were then removed and processed for electron microscopy.

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