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Background: Our knowledge about longitudinal predictors of atopy is limited., Objectives: To describe changes in skin test reactivity during a 3-year period in a sample of adolescents and adults with atopic disease and to identify factors of importance for these changes., Methods: From a sample of 1,186 candidates, ranging in age from 14 to 44 years, who in a screening questionnaire reported symptoms suggestive of respiratory allergic disease, 344 individuals with a positive skin prick test (SPT) result concomitantly with asthma and/or rhinitis were clinically studied on 2 occasions, 3 years apart., Results: In total, 134 (39% of the study population) had a different SPT result at follow-up compared with baseline, based on a diagnostic criterion of 3 mm for a positive test result; 77 (22%) developed de novo sensitizations to 1 or more allergens, 45 (13%) had remission of 1 or more sensitizations, and 12 (4%) both gained and lost sensitizations. Female sex (odds ratio = 1.90 [1.02-3.57], P = .04) significantly predicted incidence of 1 or more sensitizations., Conclusions: Four of 10 adults with atopic disease will have changes in SPT status during a 3-year period, and most will gain new sensitizations. Women with atopic disease are at increased risk of developing new sensitizations compared with men.

Confocal microscopes are often used to study specimens labelled with fluorophores. A commonly used method for simultaneous recording of the distribution of multiple fluorophores is to divide the fluorescent light emitted by the specimen into different wavelength regions using dichroic and bandpass filters. These different wavelength regions are then distributed to multiple detectors. However, the broad and overlapping spectra of commonly used fluorophores often result in considerable crosstalk between channels. A new technique, intensity-modulated multiple-beam scanning (IMS) microfluorometry, can be used to reduce this cross-talk substantially. The IMS technique is implemented with two laser beams of different wavelengths, intensity-modulated at different frequencies, which illuminate the specimen simultaneously. The two laser wavelengths predominantly excite one fluorophore each. Fluorescent light from the specimen is divided into two wavelength regions (red and green) which are detected by two photomultiplier tubes. The output signals from the photomultiplier tubes are connected to lock-in amplifiers. The effect of using modulated laser beams, in combination with lock-in amplifiers, is strongly to reduce cross-talk between the channels. The performance of the IMS technique using various types of specimen is compared with the results obtained using the conventional multi-detector method.

In a scanning laser microscope detecting fluorescent light from the specimen, the depth-discriminating property of confocal scanning has been used to carry out optical slicing of a thick specimen. The recorded digital images constitute a three-dimensional raster covering a volume of the specimen. The specimen has been visualized in stereo and rotation by making look-through projections of the digital data in different directions. The contrast of the pictures has been enhanced by generating the gradient volume. This permits display of the border surfaces between regions instead of the regions themselves.

Optical serial sectioning based on the depth-discriminating ability of confocal laser scanning can be combined with digital image processing to realize fast and easy-to-use 3-D microscopy. A great advantage as compared with traditional methods, e.g., using a microtome, is that the specimen is left undamaged. An account is given of an instrument designed for this purpose and of feasibility studies that have been carried out to assess the usefulness of the method in fluorescence microscopy.

Native fully hydrated fibrin gels formed at different fibrinogen and thrombin concentrations and at different ionic strengths were studied by confocal laser 3D microscopy, liquid permeation and turbidity. The gels were found to be composed of straight rod-like fiber elements that often came together at denser nodes. In gels formed at high fibrinogen concentrations, or with high amounts of thrombin, the spaces between the fibers decreased, indicating a decrease of gel porosity. The fiber strands were also shorter. Gel porosity decreased dramatically in gels formed at the high ionic strengths. Shorter fibers were observed and fiber swelling occurred at ionic strengths above 0.24. Quantitative parameters for gel porosity, fiber mass/length ratio and diameter were also derived by liquid permeation and turbidometric analyses of the gels. Permeation analysis showed that gel porosity (measured as Ks) decreased in gels formed at higher fibrin and thrombin concentrations in agreement with the porosity observed by microscopy. The turbidometric analysis showed good agreement with the permeation data for gels formed at various thrombin concentrations, but supported the permeation data more poorly in gels formed at different fibrinogen concentrations, especially above 2.5 mg/ml. Turbidometric analysis showed that the fiber mass/length ratio and diameter decreased in gels formed at ionic strength up to 0.24, as was seen in the permeation study. However, at higher ionic strengths swelling of the fibers was suggested from the gel turbidity data and this was also indicated by microscopy. These findings are discussed in relation to previous hydrodynamic and electron microscopic studies of fibrin gels.