Banding patterns and late replication in HeLa cells

1973 ◽  
Vol 19 (2) ◽  
Author(s):  
R. Czaker
Keyword(s):  
Hela Cells ◽  
Late Replication ◽  
Banding Patterns

CHROMOSOME STRUCTURE IN CHILOCORUS (COLEOPTERA: COCCINELLIDAE). II. THE ASYNCHRONOUS REPLICATION OF CONSTITUTIVE HETEROCHROMATIN

1976 ◽  
Vol 18 (1) ◽  
pp. 85-91 ◽  
Author(s):  
T. J. Ennis
Keyword(s):  
Chromosome Banding ◽  
Chromosome Structure ◽  
Constitutive Heterochromatin ◽  
Chromosome Replication ◽  
Data Models ◽  
Late Replication ◽  
Banding Patterns ◽  
Asynchronous Replication ◽  
Chilocorus Stigma

Chromosome replication has been analysed in four species of Chilocorus. In C. orbus Csy., C. tricyclus Smith, and C. hexacyclus Smith, centric regions of all chromosomes are last to replicate, preceded in order by heterochromatic arms and euchromatic arms. In C. stigma Say, very late replication of centric regions can be detected only in otherwise wholly euchromatic chromosomes (= monophasics); in chromosomes with one arm heterochromatic (= diphasics), these arms are last to replicate. Based on pachytene bivalent morphology and chromosome banding patterns, and supported by autoradiographic data, models are presented for the general organisation of Chilocorus chromosomes. All chromosomes in the first three species are subdivided into euchromatic arm, centric heterochromatin, and either a second euchromatic arm (monophasics) or a heterochromatic arm (diphasics). Chilocorus stigma diphasics apparently lack distinct centric organisation, and are therefore divided into euchromatic and heterochromatic arms only.

scholarly journals Characterization of the late replication banding patterns in barley chromosomes.

1997 ◽  
Vol 72 (3) ◽  
pp. 125-130 ◽  
Author(s):  
Sakurako Uozu ◽  
Tetsuya Nakazaki ◽  
Takatoshi Tanisaka
Keyword(s):  
Late Replication ◽  
Banding Patterns ◽  
Replication Banding

Platinum Compounds as Stains for Electron Microscopy

1974 ◽  
Vol 32 ◽  
pp. 230-231
Author(s):  
S. K. Aggarwal ◽  
P. McAllister ◽  
R. W. Wagner ◽  
B. Rosenberg
Keyword(s):  
Electron Microscopy ◽  
Hela Cells ◽  
Uranyl Acetate ◽  
Sarcoma 180 ◽  
Platinum Compounds ◽  
Kb Cells ◽  
En Bloc ◽  
Human Lymphoma ◽  
Ultrathin Sections ◽  
Blue Coloration

Uranyl acetate has been used as an electron stain for en bloc staining as well as for staining ultrathin sections in conjunction with various lead stains (Fig. 1). Present studies reveal that various platinum compounds also show promise as electron stains. Certain platinum compounds have been shown to be effective anti-tumor agents. Of particular interest are the compounds with either uracil or thymine as one of the ligands (cis-Pt(II)-uracil; cis-Pt(II)-thymine). These compounds are amorphous, highly soluble in water and often exhibit an intense blue coloration. These compounds show enough electron density to be used as stains for electron microscopy. Most of the studies are based on various cell lines (human AV, cells, human lymphoma cells, KB cells, Sarcoma-180 ascites cells, chick fibroblasts and HeLa cells) while studies on tissue blocks are in progress.

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.

Use of Uranium-Labeled Antibodies for Electron Microscopic Localization of Various Antigens in Mammalian Cells

1967 ◽  
Vol 25 ◽  
pp. 136-137
Author(s):  
J. P. Petrali ◽  
E. J. Donati ◽  
L. A. Sternberger
Keyword(s):  
Endoplasmic Reticulum ◽  
Hela Cells ◽  
Mammalian Cells ◽  
Specific Antiserum ◽  
Electron Microscopic ◽  
E Coli ◽  
Cytoplasmic Membranes ◽  
Viral Progeny ◽  
Ultrathin Sections ◽  
Electron Microscopic Localization

Specific contrast is conferred to subcellular antigen by applying purified antibodies, exhaustively labeled with uranium under immunospecific protection, to ultrathin sections. Use of Seligman’s principle of bridging osmium to metal via thiocarbohydrazide (TCH) intensifies specific contrast. Ultrathin sections of osmium-fixed materials were stained on the grid by application of 1) thiosemicarbazide (TSC), 2) unlabeled specific antiserum, 3) uranium-labeled anti-antibody and 4) TCH followed by reosmication. Antigens to be localized consisted of vaccinia antigen in infected HeLa cells, lysozyme in monocytes of patients with monocytic or monomyelocytic leukemia, and fibrinogen in the platelets of these leukemic patients. Control sections were stained with non-specific antiserum (E. coli).In the vaccinia-HeLa system, antigen was localized from 1 to 3 hours following infection, and was confined to degrading virus, the inner walls of numerous organelles, and other structures in cytoplasmic foci. Surrounding architecture and cellular mitochondria were unstained. 8 to 14 hours after infection, antigen was localized on the outer walls of the viral progeny, on cytoplasmic membranes, and free in the cytoplasm. Staining of endoplasmic reticulum was intense and focal early, and weak and diffuse late in infection.

Exogeneous factors are not necessary in attachment, spreading and polarization of hela cells

1978 ◽  
Vol 36 (2) ◽  
pp. 310-311
Author(s):  
W. Liebrich
Keyword(s):  
Hela Cells ◽  
Calf Serum ◽  
Glass Tube ◽  
Serum Free Medium ◽  
Nacl Solution ◽  
Free Medium ◽  
Minimum Essential Medium ◽  
Serum Free ◽  
All Solutions ◽  
Glass Support

HeLa cells were grown for 2-3 days in EAGLE'S minimum essential medium with 10% calf serum (S-MEM; Seromed, München) and then incubated for 24 hours in serum free medium (MEM). After detaching the cells with a solution of 0. 14 % EDTA and 0. 07 % trypsin (Difco, 1 : 250) they were suspended in various solutions (S-MEM = control, MEM, buffered salt solutions with or without Me++ions, 0. 9 % NaCl solution) and allowed to settle on glass tube slips (Leighton-tubes). After 5, 10, 15, 20, 25, 30, 1 45, 60 minutes 2, 3, 4, 5 hours cells were prepared for scanning electron microscopy as described by Paweletz and Schroeter. The preparations were examined in a Jeol SEM (JSM-U3) at 25 KV without tilting.The suspended spherical HeLa cells are able to adhere to the glass support in all solutions. The rate of attachment, however, is faster in solutions without serum than in the control. The latter is in agreement with the findings of other authors.

Human Adenovirus Uptake by Preirnplantation Mouse Embryos

1972 ◽  
Vol 30 ◽  
pp. 268-269
Author(s):  
D. G. Chase ◽  
W. Winters ◽  
L. Piko
Keyword(s):  
Hela Cells ◽  
Human Adenovirus ◽  
Mouse Embryos ◽  
Equilibrium Density ◽  
Cell Stage ◽  
Virus Attachment ◽  
Preimplantation Mouse Embryos ◽  
Embryo Culture Medium ◽  
Preimplantation Mouse ◽  
Mouse Cells

Although the outlines of human adenovirus entry and uncoating in HeLa cells has been clarified in recent electron microscope studies, several details remain unclear or controversial. Furthermore, morphological features of early interactions of human adenovirus with non-permissive mouse cells have not been extensively documented. In the course of studies on the effects of human adenoviruses type 5 (AD-5) and type 12 on cultured preimplantation mouse embryos we have examined virus attachment, entry and uncoating. Here we present the ultrastructural findings for AD-5.AD-5 was grown in HeLa cells and purified by successive velocity gradient and equilibrium density gradient centrifugations in CsCl. After dialysis against PBS, virus was sedimented and resuspended in embryo culture medium. Embryos were placed in culture at the 2-cell stage in Brinster's medium.

Advances in imaging SIMS studies of stained and BrdU-labelled human metaphase chromosomes

1995 ◽  
Vol 53 ◽  
pp. 932-933
Author(s):  
R. Levi-Setti ◽  
J. M. Chabala ◽  
R. Espinosa ◽  
M. M. Le Beau
Keyword(s):  
High Performance ◽  
Secondary Ion Mass Spectrometry ◽  
Analytical Sensitivity ◽  
Metaphase Chromosomes ◽  
Banding Patterns ◽  
Sector Mass Spectrometer ◽  
Staining Methods ◽  
The University ◽  
Secondary Ion ◽  
Better Than

We have shown previously that isotope-labelled nucleotides in human metaphase chromosomes can be detected and mapped by imaging secondary ion mass spectrometry (SIMS), using the University of Chicago high resolution scanning ion microprobe (UC SIM). These early studies, conducted with BrdU- and 14C-thymidine-labelled chromosomes via detection of the Br and 28CN- (14C14N-> labelcarrying signals, provided some evidence for the condensation of the label into banding patterns along the chromatids (SIMS bands) reminiscent of the well known Q- and G-bands obtained by conventional staining methods for optical microscopy. The potential of this technique has been greatly enhanced by the recent upgrade of the UC SIM, now coupled to a high performance magnetic sector mass spectrometer in lieu of the previous RF quadrupole mass filter. The high transmission of the new spectrometer improves the SIMS analytical sensitivity of the microprobe better than a hundredfold, overcoming most of the previous imaging limitations resulting from low count statistics.

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