Journal of electron microscopy technique最新文献

The surface protein of the archaebacterium Pyrodictium occultum forms two-dimensional periodic arrays of extremely poor order. Two variants of correlation averaging have been applied in order to retrieve the unit cell structure from electron micrographs of negatively stained samples: straightforward correlation averaging correcting for lateral displacements only and a more elaborate approach, including a partial compensation for rotational disorder. Surprisingly, both routes yield virtually identical structures. Inclusion of molecular motifs from highly disordered domains, which are rejected in the "straightforward" approach, appears not to improve resolution, possibly because the high local strain tends to distort the individual molecules.

The aim of this presentation is to draw attention to the problems inherent in evaluating the ultrastructure of percutaneous renal biopsies and to discuss some of the special techniques which are useful in this area. It is important to realize that the ultrastructure as it appears in this kind of material does not necessarily reflect conditions in vivo. Comparison with suitable reference material may, however, permit reliable conclusions in terms of pathological diagnosis and pathogenesis. It is advocated that purely qualitative methods, which until now have predominated in ultrastructure work with renal biopsies, be replaced by morphometry and semiquantitative methods when it is possible and practical to do so in any research situation.

Immunocytochemistry of the kidney is a unique method to integrate physiology, biochemistry, and molecular biology with morphology. Both mono- and polyclonal antibody reagents are useful and each has advantages and disadvantages. Specificity with low background is of the greatest importance. Tissue preparation techniques depend on the antigen being studied as well as the methodology to be used. Pre- and postembedding techniques combined with visualization with peroxidase reaction products or with particulate markers such as ferritin and gold must be chosen for each individual circumstance to be studied. Important applications in the kidney have included studies of glomerular antigens, specific transport proteins, and segment-specific antigens of unknown specificity. Future utilization of this technique with new molecular probes will greatly enhance our knowledge of the biology of the kidney.

The tubular epithelial cells located in the renal medulla are normally working in a hypoxic milieu. In isolated rat kidneys perfused with a cell-free medium, the medullary thick ascending limbs of Henle's loop are selectively and reproducibly injured by the imbalance between oxygen demand and supply in this area. Hypoxic lesions rapidly progress from reversible to irreversible forms of cell damage. Reversible injury consists of chromatin margination and mitochondrial swelling, which can disappear upon restoration of an adequate balance of oxygenation. Irreversible injury consists of nuclear pyknosis and cytoplasmic fragmentation, lesions which persist after re-oxygenation or even progress to cell death. Reversible and irreversible phases of hypoxic injury in this distal tubule segment are comparable to, but different from, those previously defined for the proximal tubule.

Methods for the establishment and growth of renal cell types in culture are reviewed, with emphasis on current trends. General techniques available for the isolation and culture of glomerular cells have progressed from explant to enzyme dissociation and cloning techniques. The growth characteristics and properties of cultured glomerular endothelial, epithelial, mesangial, and bone-marrow-derived cells are discussed. Studies are described in which cultures of contractile mesangial cells have led to an elucidation of their role both in normally functioning glomeruli and in disease states. Renal tubule culture techniques also have progressed from mixed tissue explants and cell isolates to fractionation of enriched tubule populations and growth of specific, individually microdissected proximal convoluted, proximal straight, thick ascending limb of Henle's loop, and collecting tubules. The differentiated tubule epithelial-specific properties of such primary cultures are discussed in relation to those of permanently growing cell lines such as MDCK and LLC-PK1. Renal tubule cultures will be invaluable for the study of the role of hormones and extracellular matrix in epithelial growth and polarity of normal structure and function. In addition, in vitro models of cultured renal tubules have been established to study the effects of age, nephrotoxins, and anoxic injury.

Defects in cell-mediated immunity caused by infection with the human immunodeficiency virus (HIV) render AIDS patients particularly susceptible to fungal pathogens. Signs and symptoms of serious infection may be nonspecific, and early diagnosis and institution of antifungal therapy is essential to decrease morbidity and mortality in this patient population. In a symptomatic individual, invasive procedures are often required to establish a microbiologic diagnosis, and histopathologic examination of tissue by light and electron microscopy is often the first indication of a serious fungal infection in an AIDS patient.