Confocal Laser Microscopy Studies Research Articles

Immunocytochemical study of microvilli in a gastric carcinoma-derived cell line.

To study the structural components of microvilli of the KATO-III cell, we used anti-villin, -ezrin, and anti-MVM (microvillous membrane prepared against mouse intestinal microvilli) antibodies. Villin and ezrin cross-link actin bundles of microvilli such as those in the small intestine and renal proximal cells. Electromicroscopically, the cytoskeletal core of microvilli of the KATO-III cell was constituted of actin-filament bundles. The anti-villin antibody but not anti-ezrin antibody reacted with the KATO-III cell as demonstrated by FITC-immunofluorescence and PAP-staining. Anti-villin, anti-MVM, but not anti-ezrin antibody, reacted with the KATO-III cell surface and with intracellular materials. Western blot analysis using anti-villin and anti-MVM antibodies revealed proteins of approximately 95 kDa (villin), and 15 kDa in the microsomal membrane fractions of KATO-III cells, respectively. Immunocytochemical and confocal laser microscopic studies showed that the cell-surface and the intracellular microcysts of KATO-III cells were preferentially decorated by anti-villin and anti-MVM antibodies. These data suggested that some actin-binding proteins, such as villin, were localized at the cell surface and on some of the intracellular cytoplasmic structures of the KATO-III cell.

A confocal laser microscopic study of the actin and actin-binding proteins (ABP) network in human cultured Kupffer cells

A confocal laser microscopic study of the actin and actin-binding proteins (ABP) network in human cultured Kupffer cells

Immunolocalization of laminin and type IV collagen in the basement membrane of uninvolved psoriatic skin. A confocal scanning laser microscopic study

Immunolocalization of laminin and type IV collagen in the basement membrane of uninvolved psoriatic skin. A confocal scanning laser microscopic study

Cytological characterization of isolated sperm cells of perennial ryegrass (Lolium perenne L.)

The cytoplasmic content and the distribution of intramembrane particles (IMPs) of the plasma membrane of isolated sperm cells of perennial ryegrass (Lolium perenne L.) have been characterized using flow cytometry, transmission electron microscopy, confocal scanning laser microscopy and freeze-fracture studies. The isolated haploid sperm cells contain a variety of cell organelles with the exception of microtubules. Proplastids and plastids with starch were observed, although only rarely. Vacuoles containing remnants of organelles and stacked lamellae of endoplasmic reticulum with cytoplasmic inclusions were observed frequently, indicating that autophagy takes place. The number of mitochondria varies from 11 to 26 with an average of 17. Generally, the nucleus has a lobed shape and displays various interphasic stages of chromatin condensation. The analysis of the number of mitochondria and the nuclear state did not show evidence of sperm cell dimorphism. The cytological variability observed, could be explained by differences in developmental stages already present in vivo at the moment of isolation. No correlation between the number of mitochondria and the nuclear cross-sectioned area and/or the condensation state of the chromatin could be found. The density of intramembrane particles of the plasma membrane on the exoplasmic fracture face is more than twice that on the protoplasmic fracture face. That is the opposite of what was found for sporophytic cells of perennial ryegrass. These results are discussed in relation to the potential use of these cells for biotechnology and developmental studies.

NMDA increases phosphorylated tau immunoreactivity in primary neuronal cultures: a confocal laser microscopic study : P. Couratier, M. Lesort, P. Sindou, F. Esclaire, J. Hugon Unité de Neurobiologie Cellulaire - Lab. d'Histologie - Faculté de Médecine - 2 rue du Dr Marcland 87025 LIMOGES Cédex FRANCE

NMDA increases phosphorylated tau immunoreactivity in primary neuronal cultures: a confocal laser microscopic study : P. Couratier, M. Lesort, P. Sindou, F. Esclaire, J. Hugon Unité de Neurobiologie Cellulaire - Lab. d'Histologie - Faculté de Médecine - 2 rue du Dr Marcland 87025 LIMOGES Cédex FRANCE

A confocal laser microscopic study of enkephalin-immunoreactive appositions onto physiologically identified neurons in the rostral ventromedial medulla

Neurons in the rostral ventromedial medulla (RVM) are important in the opioid modulation of dorsal horn nociceptive transmission. Systemically administered morphine inhibits one class of RVM cells, the on-cells; excites a second class of RVM cells, the off-cells; and has no effect on a third class, neutral cells. In contrast, iontophoretic application of morphine inhibits on-cells but does not alter the activity of either off- or neutral cells. The present study addresses whether the differential sensitivity to exogenous opioids is correlated with a differential termination pattern onto the three classes of RVM neurons by afferents containing endogenous opioids. Intracellular recordings were made from RVM neurons in rats under light halothane anesthesia. Physiologically characterized neurons were injected with Neurobiotin and then subsequently visualized with a Texas red fluorophore. Thick (50 microns) sections containing labeled RVM cells were processed for enkephalin immunoreactivity (ENK-IR) using an FITC fluorophore and then optically sectioned at 1.5 micron intervals using a dual-channel confocal laser scanning microscope. ENK-IR appositions were found on the somata and dendrites of all on-cells. Although ENK-IR varicosities were also apparently apposed to off- and neutral cells, the density of such appositions was significantly less than the density of ENK-IR appositions onto on-cells. The greater overall density of ENK-IR appositions onto on-cells was apparently due to a concentration of appositions on the soma and proximal dendrites of these neurons. These results support a model of RVM function in which endogenous opioid peptides produce an antinociceptive action by a direct inhibitory action on on-cells that facilitate nociceptive transmission. This on-cell inhibition may produce an additional antinociceptive effect by removing a possible on-cell inhibition of off-cells, which are thought to inhibit nociceptive transmission.

A Confocal scanning laser microscopic study of the developing heart

During heart development myofibrillogenesis and myocyte fiber pattern formation are critical steps in the genesis of a functional four chambered pump. The factors affecting these morphogenetic patterns are complex and include mechanical and chemical regulatory agents. The long range goal of this research is to model the effects of mechanical stimulation on heart development. In order to accomplish this goal, accurate images of the fiber patterns in the myocardium are necessary at various stages of development. In this study, we have used confocal scanning laser microscopy and immunofluorescence staining to visualize the fiber patterns and myofibrils in the developing rat heart. Sprague-Dawley rats at 10-15.5 days of gestation (Plug date was designated as day 0 with noon of plug date being designated as day 0.5) were sacrificed by lethal injection of anesthetic and their embryos were fixed in 2.0% paraformaldehyde in 0.1 M HEPES (PH 7.2) for 4-8 hours at 4°C.

Working with the confocal scanning UV‐laser microscope: specific DNA localization at high sensitivity and multiple‐parameter fluorescence

The use of fast-staining DNA-specific dyes such as DAPI or Hoechst 33342/33258 has been a major problem for confocal scanning laser microscopy (CSLM) studies of intranuclear chromatin organization. Moreover, the availability of a confocal ultraviolet scanning laser microscope configuration, which allows an excitation at wavelengths of 364 nm as well as 488, 514 and 543 nm, is a prerequisite for single as well as multiple fluorescence parameter studies, especially if these studies are concerned with the precise localization of intranuclear signals. Here we report the characteristics and application of a CSLM, which was adapted for UV-excitation and therefore enables comparison of the spatial distribution of several types of signals within one preparation. In addition to multiple-parameter studies, we have also investigated the sensitivity of the system with regard to the identification of the double-stranded DNA of lampbrush chromosome loops in germinal vesicles of amphibian oocytes.

The Confocal Scanning Laser Microscopic Study of the Pitting Corrosion on Sputter-Deposited W-Ti Alloys in 1 M NaOH Solution

The pitting corrosion of the passive films formed on the surface of sputter-deposited W-30Ti and W-53Ti alloys after immersion for 24 hours in alkaline 1 M NaOH soution at 25°C was studied by using the surface sensitive technique of the confocal scanning laser microscopic (CSLM) technique including corrosion tests and electrochemical measurements. The higher corrosion rate of the W-30Ti alloy (i.e., about 3x 10-2 mm/y) than those of the W-53Ti and W-90Ti alloys (i.e., about 4-5 x 10-3 mm/y) was mostly due to the deep type of pitting corrosion observed on the surface of the passive film formed on the W-30Ti alloy in 1 M NaOH solution. The shallow type of pits was developed on the surface of the passive film formed on the W-53Ti alloy, whereas no pitting corrosion on the titanium-rich W-90Ti alloy.Keywords: Pitting corrosion; Open circuit corrosion potential; Corrosion test; CSLM studyTribhuvan University Journal Vol. XXVI, No. 1, 2009 Page: 17-26