Identification and study of the same cell in monolayer culture by different methods of light microscopy, scanning, and transmission electron microscopy. Application for cryodamaged cells examination

Katsen, A. D.; Tregubova, N A; Andrushkevich, V. V.; Rovensky, Yu. A.

doi:10.1002/sca.4950140407

Cited by 6 publications

(10 citation statements)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…564–570], and indium foil was found to be a suitable substrate [ Physical Electronics , 1997]. Indium oxide has been used for coating of glass slides in SEM studies of biological material [ Ogura and Kuscheck , 1980; Katsen et al , 1992]. However, to our knowledge, indium has not been used hitherto in aerosol research.…”

Section: Methodsmentioning

confidence: 99%

New evidence of an organic layer on marine aerosols

et al. 2002

View full text Add to dashboard Cite

[1] An extraordinary episode of fine particles (diameter mainly <2.5 mm) occurred in Helsinki, southern Finland, at the end of February 1998. The air masses came from the North Atlantic Ocean and passed over France, Germany, and southern Scandinavia. Particles were collected during the episode as well as before and after it. Uncoated particle samples were adhered to an indium substrate and were studied by a scanning electron microscope (SEM) coupled with an energy dispersive X-ray microanalyzer (EDX). A great proportion of the particles behaved differently than aerosols previously studied by microscopic techniques. The particles (size mainly 0.5-1 mm) did not exhibit solid shape. They were ''bubbling'' or ''pulsating'' continually, enlarging in one part and shrinking in another. Some particles were broken down, especially when the beam of the electron microscope was focused on them. EDX analyses showed that the particles contained much carbon together with oxygen, sulfur, and sodium. Ion analyses by ion chromatography revealed high concentrations of sodium, sulfate, nitrate, and ammonium. The particles were identified as marine sea-salt aerosols, which had accumulated anthropogenic emissions and lost chloride during their flow through continental polluted air. The shape fluctuations and the high carbon content observed by SEM/EDX led to the conclusion that the aerosols were enclosed by an organic membrane. Direct insertion probe/mass spectrometry investigations showed remarkable amounts of fragmented aliphatic hydrocarbons, which were considered as an evidence of a lipid membrane on the surface of the particles. The impact of the posited organic film on the properties of sea-salt particles, as well as on Earth's climate, is discussed.

show abstract

Section: Methodsmentioning

confidence: 99%

New evidence of an organic layer on marine aerosols

et al. 2002

View full text Add to dashboard Cite

show abstract

“…After SEM examination, specimens were prepared for transmission electron microscopy (TEM) by embedding in Araldite 502 and removing from the substrate after polymerization [13]. Again, the diamond replica served to relocalize the cells that were to be cut with the ultramicrotome.…”

Section: Introductionmentioning

confidence: 99%

“…The postfixation was performed in three steps using 1% osmium tetroxide, 1% tannic acid and 1% uranyl acetate, a schedule especially developed for SEM [13]. After dehydration in ethanol, specimens were critical-point-dried and sputtered with gold (~5 nm thick).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Cell surface and nuclear changes during TNF-α-induced apoptosis in WEHI 164 murine fibrosarcoma cells

et al. 1998

Self Cite

View full text Add to dashboard Cite

Tumour necrosis factor (TNF)-alpha-induced apoptosis is associated with several nuclear and cell surface alterations, in particular with the condensation of chromatin and the fragmentation of the cell nucleus, formation of blebs on the cell surface and breakdown of the plasma membrane. However, there is little information about the relationship between the cell surface alterations and the nuclear changes during apoptosis. To study this, cultured WEHI cells were exposed to TNF-alpha over different time periods. The cytological changes were studied using a correlative approach, which allowed observation of the same cell consecutively under light, scanning and transmission electron microscopy. The earliest sign of cell alteration was a reduction of the number of microvilli after 15 min of TNF-alpha exposure. This reaction was reversible (reappearance of microvilli) and took place during the first hour, in which neither nuclear alterations nor plasma membrane breakdown were observed. The changes in the nucleus began with condensation of chromatin after approximately 1 h of TNF-alpha-exposure. After 4-5 h the microvilli disappeared again, particularly in areas where the formation of blebs (blebbing) was observed. Strikingly, cell surface alterations (bleb formation) were detected only in those cells that presented with condensed chromatin, and not in cells with a normal chromatin pattern, proving at least a close correlation between nuclear and cell surface changes during the process of apoptosis.

show abstract

“…The term correlative microscopy was first used with the meaning of combining different microscopic techniques with the identical sample but with adapted preparation steps for each microscopic approach (22). However, under these circumstances part of the observed structural variations result rather from different pretreatments than from biological origin (2).…”

Section: Resultsmentioning

confidence: 99%

A strategy for correlative microscopy of large skin samples: towards a holistic view of axillary skin complexity

Wilke

Wick

Keil

et al. 2007

Experimental Dermatology

View full text Add to dashboard Cite

Knowledge about the structural elements of skin and its appendices is an essential prerequisite for understanding their complex functions and interactions. The hence necessary morphological description across several orders of scale not only requires the investigation at the light microscopic level but also ultrastructural investigation, ideally on the identical sample. For a correlative and multimodal observation one unique preparation protocol is mandatory. As a compromise between sample sizes of >500 lm in diameter on the one hand and optimal preservation of antigenicity and morphology on the other, we developed a new preparation protocol that allows (i) 3D reconstruction of the resin-embedded sample by confocal light microscopy prior to (ii) direct immunolocalization of target proteins within selected sample planes by light and fluorescence microscopy or transmission electron microscopy. Alternatively, (iii) serial cryosections of the frozen sample can be taken for characterizing the sample in toto. With this unique approach we were able to fully demonstrate the structural complexity of axillary skin samples, increasing the structural resolution from 3D reconstruction of the whole gland up to ultrastructural investigations at the subcellular level. We could demonstrate that axillary sweat glands are not separately distributed, as has been assumed to date; instead, they seem to be intricately twisted into one another. This promotes the concept of a complex axillary sweat gland organ instead of single sweat gland entities.Key words: axillary skin -correlative microscopy -multimodal imaging -sweat gland Please cite this paper as: A strategy for correlative microscopy of large skin samples: towards a holistic view of axillary skin complexity. Experimental Dermatology 2008; 17: 73-80.

show abstract

Identification and study of the same cell in monolayer culture by different methods of light microscopy, scanning, and transmission electron microscopy. Application for cryodamaged cells examination

Cited by 6 publications

References 13 publications

New evidence of an organic layer on marine aerosols

New evidence of an organic layer on marine aerosols

Cell surface and nuclear changes during TNF-α-induced apoptosis in WEHI 164 murine fibrosarcoma cells

A strategy for correlative microscopy of large skin samples: towards a holistic view of axillary skin complexity

Contact Info

Product

Resources

About