Fluorospot assay: methodological analysis.

The conventional enzyme-linked immunospot (ELISPOT) technique detects only one secreted cytokine, which constitutes a major pitfall for the accurate characterization of the various T-cell subpopulations. We have therefore developed a fluorospot assay, which is a modification of the ELISPOT and is based on the use of multiple fluorescent-labeled anticytokines detection antibodies. A special automated ELISPOT reader consisting of a light microscope with incident fluorescence illumination and an integrating digital color camera has been adapted for this technique.

High resolution as a key feature to perform accurate ELISPOT measurements using Zeiss KS ELISPOT readers.

The enzyme-linked immunospot (ELISPOT) assay was originally developed for the detection of individual antibody secreting B-cells. Since then, the method has been improved, and ELISPOT is used for the determination of the production of tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma, or various interleukins (IL)-4, IL-5. ELISPOT measurements are performed in 96-well plates with nitrocellulose membranes either visually or by means of image analysis.

The vascular architecture of the chick chorioallantoic membrane: sequential quantitative evaluation using corrosion casting.

Microvascular corrosion casting was used for evaluating qualitatively and quantitatively angiogenesis in the chick chorioallantoic membrane (CAM). Series of CAMs from day 8 to 18 were examined. The density of plexus capillaries increases rapidly until day 10 and then remains constant.

Digital light microscopy: prerequisite for optimum contrast enhancement and increase of resolution.

Since the reproducible production of microscope objectives was enabled by the lens calculations of Ernst Abbe in 1872, various attempts have been made to further increase the resolution of light microscopes. Apart from the improvements on the optical side, especially the introduction of fluorescence methods, the use of digital cameras connected to computers have brought us close to the theoretical limits of optical resolution. Due to improved speed and memory capacity of modern computers mathematical methods can be applied to stored three-dimensional (3D) sequences of digital images which, in addition to just contrast and edge enhancement, may result in the case of real 3D deconvolution, even in the visualisation of structures beyond the theoretical limitations of light optical resolution.

Correlation of endoanal sonography with cross-sectional anatomy of the anal sphincter.

Background: Anal endosonography has become an important imaging method in the diagnosis of anorectal disorders. However, little information exists as to whether anal endosonography reliably defines pelvic floor structures. The aim of this study was to correlate endoanal sonography with cross-sectional anatomy and histology.