Reflection contrast microscopy for high resolution detection of3H-estradiol in ultrathin sections of human stratum corneum

Neelissen, Jan; Schrijvers, Ad H. G. J.; Junginger, Hans E.; Boddé, Harry E.

doi:10.1002/(sici)1097-0029(19991115)47:4<286::aid-jemt6>3.0.co;2-g

Cited by 2 publications

(2 citation statements)

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“…Recent promising results obtained with freezedried embedded material and the different available microanalytical techniques have been published (e.g. Burns & File, 1986;Grohovaz et al, 1996;Pezzati et al, 1997;Bücking et al, 1998Neelissen et al, 1999;. In the future, SIMS will become probably the most powerful technique for the microanalytical investigation of freeze-dried embedded material.…”

Section: Microanalytical Studiesmentioning

confidence: 99%

Freeze‐dried and resin‐embedded biological material is well suited for ultrastructure research

Edelmann

2002

Journal of Microscopy

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SummaryTransmission electron micrographs of different biological material, cryofixed, freeze-dried and embedded in Spurr's resin, in Epon, or in Lowicryl, are presented. The structure preservation obtained either without or with application of chemical fixatives after drying showed that freeze-dried embedded specimens are particularly well suited for new morphological, immunocytochemical and microanalytical studies aimed at detecting the life-like subcellular distribution of mobile macromolecules and ions. The results also indicate that the removal of cell water by freeze-drying from the areas of best cryofixation is relatively slow. Ultrathin sections of well cryofixed biological material embedded after freeze-drying in Spurr's resin or Epon reveal cellular plasma phases with very fine granularities and well defined membranes in negative contrast. This may be due to the preservation of the original structure of cellular macromolecules with a considerable amount of their hydration water. Sublimation studies with differently hydrated and cryofixed macromolecules are suggested to settle this issue.

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Section: Microanalytical Studiesmentioning

confidence: 99%

Freeze‐dried and resin‐embedded biological material is well suited for ultrastructure research

Edelmann

2002

Journal of Microscopy

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“…Deroost et al (2013) could visualise schizonts in RCM studies of blood in malaria cases. Neelissen et al (1999) described the detection of 3H estradiol in ultrathin sections of human stratum corneum. Filler & Peuker (1997) mentioned an improved detection of peripheral nerve tissue with RCM.…”

Section: Applications Of Reflection-contrast Microscopy (Rcm)mentioning

confidence: 99%

Applications of reflection‐contrast microscopy, including the sensitive detection of the results ofin situhybridisation a review

Ploem

2019

Journal of Microscopy

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Summary The observation with RCM of the reflection from reaction products produced by nonisotopic in situ hybridisation and a peroxidase staining, has recently facilitated the identification of single copy genes. RCM also reveals light microscope structures in stained ultrathin (0.1 μm) epon and lowicryl sections. In such preparations no out‐of‐focus blur can be observed. Confocal optics are thus not needed to obtain high‐quality images of ultrathin sections at the highest conventional light microscope optical resolution, and with higher image‐contrast than can be obtained by classical absorption microscopy. Such RCM images provide the same type of information as confocal scanning microscope images. A sequential ultrathin section of the same microscopic specimen can be examined with electron microscopy for a simple and accurate (CLEM) procedure. For some applications RCM can replace (CLSM). Lay Description With reflection contrast microscopy (RCM), a microscope image can be observed if micromirror‐like reflecting substances are present in the reaction product of cytochemical stains. Using an antiflex objective for RCM, the reflected incident light from the stained microscope specimen can be observed with only a small amount of unwanted stray‐light in the image background. Stray‐light is caused by reflection of incident light from the surface of the lenses in the microscope objective and in the microscope tube (using a 50% beam splitter for epi‐illumination). Several authors who compared RCM with bright‐field and fluorescence microscopy, found RCM to be a sensitive microscope method in many applications. It has been reported that RCM, using in situ hybridisation methods, can detect DNA sequences in single copy genes. In the literature is mentioned that single copy genes have an estimated mass of less than a trillionth of a gram of DNA. From stained ultrathin microscope sections with less than 0.1 μm thickness, multicoloured images with a high image contrast, optimal resolution and no out‐of‐focus blur can be observed with RCM. To achieve this, RCM uses an antiflex microscope objective, with a quarter‐wave plate mounted on the front lens in combination with a polarising filter cube inserted in the epi‐illuminator. RCM can also make use of oblique illumination, for a further increase of the image‐contrast and the resolution of the microscope image. A central stop has then to be inserted in the aperture plane of an epi‐illuminator.

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Reflection contrast microscopy for high resolution detection of3H-estradiol in ultrathin sections of human stratum corneum

Cited by 2 publications

References 22 publications

Freeze‐dried and resin‐embedded biological material is well suited for ultrastructure research

Freeze‐dried and resin‐embedded biological material is well suited for ultrastructure research

Applications of reflection‐contrast microscopy, including the sensitive detection of the results ofin situhybridisation a review

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