Coming of age: reconstruction of heterosexual HIV-1 transmission in human ex vivo organ culture systems.

Heterosexual transmission of human immunodeficiency virus-1 (HIV-1), from men to women, involves exposure to infectious HIV-1 in semen. Therefore, the cellular and molecular processes that underlie HIV-1 transmission are closely interconnected with fundamental principles of human reproductive biology. Human ex vivo organ culture systems allow experimental reconstruction of HIV-1 transmission, using human semen and premenopausal cervicovaginal mucosal tissue, with specific emphasis on the progression from exposure to development of primary HIV-1 infection. Clearly, an isolated piece of human tissue cannot duplicate the full complexity of events in natural infections, but with correct observation of conventional medical and ethical standards, there is no opportunity to study HIV-1 exposure and primary infection in young women. Human mucosal organ cultures allow direct study of HIV-1 infection in a reproducible format while retaining major elements of complexity and variability that typify community-based HIV-1 transmission. Experimental manipulation of human mucosal tissue both allows and requires acquisition of new insights into basic processes of human mucosal immunology. Expanding from the current foundations, we believe that human organ cultures will become increasingly prominent in experimental studies of HIV-1 transmission and continuing efforts to prevent HIV-1 infection at human mucosal surfaces.

Inner-shell electron spectroscopy for microanalysis.

The transmission electron energy-loss spectrum shows characteristic "edges" corresponding to the excitation of inner-shell electrons of atoms in a thin sample. Analysis of these edges provides detailed chemical, structural, and electronic data from the radiated volume. By combining electron spectroscopy and electron microscopy, this microanalytical technique can be performed in conjunction with highresolution imaging of the sample. It is shown that this approach has advantages of sensitivity, spatial resolution, and convenience over other comparable techniques.

Fluorinated molecule as a tracer: difluoroserotonin in human platelets mapped by electron energy-loss spectroscopy.

The intracellular distribution of fluorine has been delineated in human platelets incubated with 4,6-difluoroserotonin, utilizing a scanning-transmission electron microscope equipped with an energy-loss spectrometer. Discrete intracellular structures corresponding in location to dense bodies contained high concentrations of fluorine. Electron energy-loss spectroscopy, which apparently can detect less than 10(-20) gram of fluorine in an area of 10 square nonometers, can thus localize fluorinated tracer molecules with biological activity.

The formation and interpretation of defect images from crystalline materials in a scanning transmission electron microscope.

The technique of scanning transmission electron microscopy (STEM) has been employed usefully in studies of amorphous materials, and the theory of image formation and interpretation in this case has been well developed. Less attention has been given to the practical and theoretical problems associated with the use of STEM for the examination of crystalline materials. In this case the contrast mechanisms are dominated by Bragg diffraction and so they are quite different from those occurring in amorphous substances. In this paper practical techniques for the observation and interpretation of contrast from defects in crystalline materials are discussed. It is shown that whilst images of defects are obtained readily under all typical STEM operating conditions, the form of the image and the information it contains varies with the angle subtended at the specimen by the detector. If this angle is too large significant image modifications relative to the "conventional" transmission electron microscope case may occur and the resolution of the image may degrade. If this angle is too small, then signal to noise considerations make an interpretation of the image difficult. In this paper we indicate how the detector angle may be chosen correctly, and also present techniques for setting up a STEM instrument for imaging a crystalline material containing lattice defects.

Topographical contrast in the transmission electron microscope.

An adaptation of the Foucault method for topographical imaging in the transmission electron microscope is described in detail. The image contrast is produced by selection of electrons which have suffered differential phase retardations in the specimen inner potential. Surface or interface displacements produce bright or dark image contrast, and the ultimate resolution approaches that of the atomic scale. The imaging method is applied in studies of both amorphous and crystalline objects. The possibility of performing quantitative measurements is demonstrated by the estimation of the inner potential of crystalline MgO.