Embedding Free-Floating Cells and Microscopic Particles: Serum Albumin Coagulum—Epoxy Resin

Jw, Shands

doi:10.3109/10520296809115036

Cited by 30 publications

(9 citation statements)

References 3 publications

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“…After pelleting in a BSA-glutaraldehyde mixture [24], the samples were processed for acetylation [29], and epon embedded; ultrathin sections were counterstained with uranyl and lead. EDTA regressive staining, preferential for ribonucleoproteine (RNP) was performed on thin sections as previously described [3].…”

Section: Transmission Electron Microscopymentioning

confidence: 99%

Candida albicans— adriamycin interactions: ultrastructural and spectrofluorometric study of whole yeasts and spheroplasts

Bobichon

Bussy

Angiboust

et al. 1990

Biology of the Cell

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The occurrence of candidiasis in cancer patients who undergo chemotherapy requires the interrelation of Candida albicans and the antimitotic drug Adriamycin (ADM) which is well known as an intercalating agent. The whole yeasts were not affected by 2 h of contact with the drug at 10(-4) M neither for their growth curve nor for their ultrastructure, despite the presence of free ADM on their surface. Spheroplasts displayed a delay in their growth and exhibited altered nucleoli with segregation of their granular and fibrillar components. The modified emission spectrum of ADM, determined by spectrofluorometry, corresponded neither to the free ADM nor to the DNA-bound drug, but it could be related to a metabolite of the drug. The cell wall appeared to be one of the main sites for ADM resistance of Candida albicans in vitro.

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Section: Transmission Electron Microscopymentioning

confidence: 99%

Candida albicans— adriamycin interactions: ultrastructural and spectrofluorometric study of whole yeasts and spheroplasts

Bobichon

Bussy

Angiboust

et al. 1990

Biology of the Cell

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“…The cells were then rinsed three times in PBS, centrifuged and resuspended in a 10% solution of bovine serum albumin in 0.05 M Tris buffer pH 7.2, gelled by the addition of one drop of 25%0 glutaraidehyde according to the method of Shands [12]. Before fixation, freshly isolated BTG cells were washed with PBS, centrifuged and then fixed for 30 rain with a solution of 2.5%o glutaraldehyde in PBS.…”

Section: Characterization Of Isolated and Cultured Bovine Tracheal Glmentioning

confidence: 99%

“…The post-fixation was omitted for cytochemical and immunocytochemical studies. The cells were then rinsed three times in PBS, centrifuged and resuspended in a 10% solution of bovine serum albumin in 0.05 M Tris buffer pH 7.2, gelled by the addition of one drop of 25%0 glutaraidehyde according to the method of Shands [12]. The gel encapsulated pellet was then dehydrated through a graded series of ethanol and embedded in Epon.…”

Section: Characterization Of Isolated and Cultured Bovine Tracheal Glmentioning

confidence: 99%

Growth and characterization of isolated bovine tracheal gland cells in culture. Influence of a reconstituted basement membrane matrix

Benali

Dupuit

Jacquot

et al. 1989

Biology of the Cell

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We describe a method for establishing the culture of bovine tracheal submucosal gland (BTG) cells, in which we have also examined the influence of a reconstituted basement membrane matrix derived from the Engelbreth-Holm-Swarm tumor (EHS) on the growth and morphological differentiation of these cells. BTG cells have been isolated by tissue enzymatic digestion using trypsin, deoxyribonuclease I, elastase, hyaluronidase and EGTA for 1 hr at 37 degrees C. Afterwards, cells and tissue were collected by centrifugation and were incubated for 15 min with 15% newborn calf serum to inactivate the proteolytic enzymes. Enzymatic digestion using only trypsin, centrifugation and inactivation steps were repeated three times. Using this protocol, we obtained 15 +/- 4 (X 10(6] cells per g of tracheal submucosa with 72 +/- 2% (n = 5) cell viability. On microscopic observation, isolated cells were mainly composed of serous type glandular cells. Cells were cultured in a 1:1 medium of Dulbecco's Modified Eagle's/Ham's F12 supplemented with 10% fetal calf serum and subcultured in either plastic flasks or flasks coated with EHS matrix. On the plastic, the BTG cells exhibited at confluency an epithelioid appearance. They stained positively with the immunofluorescent anticytokeratin antibody and contained PAS-staining granules. By electron microscopy, lactoferrin, a protein marker specific to the serous cells, was demonstrated immunocytochemically in small secretory vesicles. BTG cells cultured on EHS matrix revealed a significantly increased growth in comparison to those cultured on plastic. In post-confluent culture of BTG cells on EHS matrix, we observed numerous dome-like structures formed by differentiated cells which were joined together around luminal spaces.

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“…Cell suspensions or minute organisms can be embedded in agar (Ryter & Kellenberger, 1958;Hirsch & Fedorko, 1968;Salyeav, 1968;Saikawa & Kobayashi, 1974;Trett, 1980). agarose (Seed, 1980), fibrin clots (Furtado, 1970) or bovine serum albumen (Shands, 1968) for electron microscopy. Although larger blocks (approx, 1 mrn3) of the embedding materials are easier to process than the original specimens, some embedding materials, such as agar and agarose, become nearly invisible when immersed in fluid and therefore they can be lost in discard fluid as easily as the original specimen.…”

Section: Introductionmentioning

confidence: 99%

A simple filter system for processing small or transparent specimens

Cribb

Zhu

1994

Journal of Microscopy

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Embedding Free-Floating Cells and Microscopic Particles: Serum Albumin Coagulum—Epoxy Resin

Cited by 30 publications

References 3 publications

Candida albicans— adriamycin interactions: ultrastructural and spectrofluorometric study of whole yeasts and spheroplasts

Candida albicans— adriamycin interactions: ultrastructural and spectrofluorometric study of whole yeasts and spheroplasts

Growth and characterization of isolated bovine tracheal gland cells in culture. Influence of a reconstituted basement membrane matrix

A simple filter system for processing small or transparent specimens

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