The Reactions of Osmium Tetroxide With Lipids and Other Compounds

Khan, Abdul Aziz; Riemersma, Jelle C.; Booij, H. L.

doi:10.1177/9.5.560

Cited by 38 publications

(11 citation statements)

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“…3c ). After entering the membrane, OsO 4 can form cyclic osmate esters by covalent binding to unsaturated membrane lipids 11 12 13 ( Fig. 3a , step 2).…”

Section: Resultsmentioning

confidence: 99%

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Large-volume en-bloc staining for electron microscopy-based connectomics

2015

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Large-scale connectomics requires dense staining of neuronal tissue blocks for electron microscopy (EM). Here we report a large-volume dense en-bloc EM staining protocol that overcomes the staining gradients, which so far substantially limited the reconstructable volumes in three-dimensional (3D) EM. Our protocol provides densely reconstructable tissue blocks from mouse neocortex sized at least 1 mm in diameter. By relaxing the constraints on precise topographic sample targeting, it makes the correlated functional and structural analysis of neuronal circuits realistic.

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“…3c ). After entering the membrane, OsO 4 can form cyclic osmate esters by covalent binding to unsaturated membrane lipids 11 12 13 ( Fig. 3a , step 2).…”

Section: Resultsmentioning

confidence: 99%

“…3a , step 2). However, these intermediates slowly undergo dihydroxylation 11 14 ( Fig. 3a , step 3), with the effect of Os leaving the membrane again, yielding [OsO 2 (OH) 4 ] 2− , a water soluble Os compound at oxidation level VI.…”

Section: Resultsmentioning

confidence: 99%

Large-volume en-bloc staining for electron microscopy-based connectomics

2015

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“…The work of Bahr (3) indicates that cholesterol is not rapidly colored by osmium. On the other hand, osmium tetroxide unites in a 1:1 molar ratio with fatty acids exhibiting a single unsaturation and in higher ratios to those with more sites of unsaturation (20). It is possible, therefore, that the images seen in the micrographs obtained from this material give some indication of the composition of the droplets as well as the droplet size.…”

Section: Fmvrementioning

confidence: 99%

Observations on the Fine Structure of Lutein Cells

Enders

Lyons

1964

The Journal of Cell Biology

110

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Corpus luteum formation was induced in 26-day-old rats which were subsequently hypophysectomized and injected with mammotrophic hormone (MH, LTH). Sections of corpora lutea from these animals were examined with the electron microscope and compared with similarly prepared (Caulfield's fixed, Araldite embedded) corpora from normal pregnancy and from controls, the latter consisting of corpora prior to hypophysectomy and corpora from uninjected rats 7 to 14 days after hypophysectomy. Lutein cells from corpora lutea of injected animals and of normal pregnancy are characterized by abundant, tortuous, tubular agranular endoplasmic reticulum and by mitochondria, many of which are disc-shaped with dense matrices and both villiform and lamclliform cristae. The endoplasmic reticulum is most abundant in lutein cells from pregnant animals, in which cells it is in the form of thin, highly tortuous tubules. The form of the lipid droplets seen in cells of stimulated animals varies greatly. Marginal foldings of the lutein cells on the perivascular space were found in all instances. Lutein cells from hypophysectomized animals have a less highly developed agranular endoplasmic reticulum. The mitochondria have irregular outlines and a relatively lucid matrix. The lipid droplets in these cells show less tendency to be extracted, but are not so large or abundant as in the cells of onset controls. Granules believed to contain lipid pigments are common in the lutein cells of these control animals. It is suggested that lutein cells from corpora lutea which are actively secreting progesterone may be readily distinguished from lutein cells from non-active corpora by means of the multiple characteristics enumerated. It is further suggested that mammotrophic hormone has a general effect on the metabolism of lutein cells rather than solely affecting a specific organelle, the abundance or composition of which may be the limiting factor in the production of progesterone.In a previous paper active lutein cells from three species were studied. It was found that these lutein cells were characterized by the presence of marginal folding on the perivascular space, extensive agranular endoplasmic reticulum, and abundant mitochondria with numerous villiform cristae. It is the intention of the present study to examine the structure of inactive as well as active corpora lutea.Since the early studies on a corpus luteumstimulating factor (2, 13, 15) and the dependence of the corpus luteum of the rat on this luteotrophic factor (1,14), the function of mammotrophic

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“…The first, of two, osmium incubations uses reduced osmium, which encourages more membrane/lipid staining. Osmium will mainly bind to C=C bonds in unsaturated fats (Khan et al, 1961), providing a strong membrane stain. Subsequently adding the mordant thiocarbohydrazide, which binds the osmium left by the first incubation, allows the binding of further osmium in a subsequent incubation therefore enhancing the stain (Seligman et al, 1966).…”

Section: Sample Preparationmentioning

confidence: 99%

Serial block face scanning electron microscopy in cell biology: Applications and technology

Smith

Starborg

2019

Tissue and Cell

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Three-dimensional electron microscopy (3DEM) is an imaging field containing several powerful modalities such as serial section transmission electron microscopy and electron tomography. However, large-scale 3D studies of biological ultrastructure on a cellular scale have historically been hampered by the difficulty of available techniques. Serial block face scanning electron microscopy (SBFSEM) is a 3DEM technique, developed in 2004, which has greatly increased the reliability, availability and throughput of 3DEM. SBFSEM allows for 3D imaging at resolutions high enough to resolve membranes and small vesicles whilst having the capability to collect data with a large field of view. Since its introduction it has become a major tool for ultrastructural investigation and has been applied in the study of many biological fields, such as connectomics, cellular and matrix biology. In this review, we will discuss biological SBFSEM from a technical standpoint, with a focus on cellular applications and also subsequent image analysis techniques.

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The Reactions of Osmium Tetroxide With Lipids and Other Compounds

Cited by 38 publications

References 0 publications

Large-volume en-bloc staining for electron microscopy-based connectomics

Large-volume en-bloc staining for electron microscopy-based connectomics

Observations on the Fine Structure of Lutein Cells

Serial block face scanning electron microscopy in cell biology: Applications and technology

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