Use of scanning electron microscopy for the evaluation of intestinal epithelium integrity

Propõe-se, neste trabalho, o uso de microscopia eletrônica de varredura para avaliar perda de epitélio intestinal da mucosa intestinal de pintos machos e fêmeas submetidos a prolongado jejum pós-eclosão de água e ração. Dois segmentos do duodeno, jejuno e íleo por ave foram coletados e processados pelo método de rotina para microscopia eletrônica de varredura. Seis diferentes graus de perda de epitélio intestinal foram determinados: grau 0, vilos normais, sem extrusão; grau 1, vilos com pequenos pontos de extrusão; grau 2, vilos com perda de epitélio no ápice; grau 3, vilos com perda de epitélio na região apical; grau 4, vilos com perda de epitélio em sua metade superior; grau 5: vilos sem epitélio; grau 6: vilo quebrado. As três regiões intestinais das fêmeas apresentaram aproximadamente 90% de seus vilos normais (graus 0 e 1), enquanto nos machos ocorreu 38% de vilos normais no duodeno e jejuno e 85% no íleo. Além disso, machos apresentaram graus mais acentuados de perda de epitélio (graus 3, 4, 5 e 6) que as fêmeas (grau 3). Os dados indicaram que a mucosa intestinal de pintos machos é mais sensível a prolongado jejum pós-eclosão que a de pintos fêmeas. A microscopia eletrônica de varredura pode ser usada como um método de rotina seguro para a caracterização e quantificação de perda de epitélio intestinal.

Observation of the embryonic development in Pseudoplatystoma coruscans (Siluriformes: Pimelodidae) under light and scanning electron microscopy

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Early development of Astronotus ocellatus under stereomicroscopy and scanning electron microscopy

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Mechanism of infection and colonization of Rhipicephalus sanguineus eggs by Mertarhizium anisopliae as revealed by scanning eletron microscopy and histopathology

O presente trabalho teve como objetivo verificar a forma de penetração do fungo Metarhizium anisopliae em ovos do carrapato Rhipicephalus sanguineus, assim como as lesões infringidas no interior do ovo. A aderência e penetração do fungo foram estudadas por meio da microscopia eletrônica de varredura e a ação do fungo nos tecidos internos avaliada em secções histológicas convencionais. Para observação destes eventos, realizaram-se infecções experimentais em 11 grupos de ovos do R. sanguineus contendo 25 mg cada. Os ovos foram banhados durante 3 minutos, sob agitação manual, em suspensão com concentração de 10(8) conídios/mL. Nos grupos controle o banho foi realizado apenas no veículo da suspensão. Os ovos foram processados para análise histopatológica e microscopia eletrônica de varredura nos seguintes tempos após a infecção: 1 e 18h, e um, dois, três, quatro, cinco, seis, sete, nove e onze dias. Observou-se grande germinação de conídios em 67% dos ovos 18h após a inoculação e o fungo penetrou em 92,6% dos ovos 5 dias após a infecção. A extrusão do patógeno ocorreu em 87% dos ovos 7 dias após a infecção, chegando a 100% no 9º dia. Nas análises histopatológicas não foram observadas lesões dignas de nota, porem deve-se ressaltar que houve significativa redução (53,9%) na eclosão a partir dos ovos infectados.

Interaction of small amounts of bovine serum albumin with phospholipid monolayers investigated by surface pressure and atomic force microscopy

The influence of small amounts of bovine serum albumin (BSA) (nM concentration) on the lateral organization of phospholipid monolayers at the air-water interface and transferred onto solid substrates as one-layer Langmuir-Blodgett (LB) films was investigated. The kinetics of adsorption of BSA onto the phospholipid monolayers was monitored with surface pressure isotherms in a Langmuir trough, for the zwitterionic dipalmitoylphosphatidyl ethanolamine (N,N-dimethyl-PE) and the anionic dimyristoylphosphatidic acid (DMPA). A monolayer of N,N-dimethyl-PE or DMPA incorporating BSA was transferred onto a solid substrate using the Langmuir-Blodgett technique. Atomic force microscopy (AFM) images of one-layer LB films displayed protein-phospholipid domains, whose morphology was characterized using dynamic scaling theories to calculate roughness exponents. For DMPA-BSA films the surface is characteristic of self-affine fractals, which may be described with the Kardar-Parisi-Zhang (KPZ) equation. on the other hand, for N,N-dimethyl-PE-BSA films, the results indicate a relatively flat surface within the globule. The height profile and the number and size of globules varied with the type of phospholipid. The overall results, from kinetics of adsorption on Langmuir monolayers and surface morphology in LB films, could be interpreted in terms of the higher affinity of BSA to the anionic DMPA than to the zwitterionic N,N-dimethyl-PE. Furthermore, the effects from such small amounts of BSA in the monolayer point to a cooperative response of DMPA and N,N-dimethyl-PE monolayers to the protein. (c) 2005 Elsevier B.V. All rights reserved.

Description of eggs and nymphal instars of Triatoma baratai Carcavallo & Jurberg, 2000 based on optical and scanning electron microscopy (Hemiptera: Reduviidae: Triatominae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Morphology of the eggs of the genus Belminus (Hemiptera: Reduviidae: Triatominae) by optical and scanning electron microscopy

The external morphology of eggs of the species Belminus corredori Galvao & Angulo, 2006, Belminus herreri Lent & Wygodzinsky, 1979 and Belminus ferroae Sandoval, Pabon, Jurberg & Galvao, 2007 is described for the first time using scanning electron microscopy (SEM). Females lay one egg at a time; the shape of the egg is oval with lateral flattening, a rounded caudal area and a true and simple convex operculum. The average size of the egg among these species is 0.51-0.58 mm in length and 0.36-0.4 mm in width. As a generic character we highlight the discovery of a chorion rim with one to four grooves always joined to the micropyles. All species present a clear polygonal pattern only in the operculum and in the egg's cephalic region, while the median and caudal regions show a smooth appearance, features that are shared only with the eggs of the genus Alberprosenia. A discriminant analysis of the polygonal design of the operculum proved to be useful for the identification of the species.

Improved laboratory safety by decontamination of unstained sputum smears for acid-fast microscopy

Tubercle bacilli may survive in unstained heat-fixed sputum smears and may be an infection risk to laboratory staff. We compared the effectiveness of 1% and 5% sodium hypochlorite, 5% phenol, 2% glutaraldehyde, and 3.7% formalin in killing Mycobacterium tuberculosis present in smears prepared from 51 sputum samples. The smears were decontaminated by the tube and slide techniques. Phenol at 5%, glutaraldehyde at 2%, and buffered formalin at 3.7% for 1 min (tube technique) or for 10 min (slide technique) were effective in decontaminating sputum smears and preserved cell morphology and quantitative acid-fast microscopy results.

Direct observation of oxidative stress on the cell wall of Saccharomyces cerevisiae strains with atomic force microscopy

We imaged pores on the surface of the cell wall of three different industrial strains of Saccharomyces cerevisiae using atomic force microscopy. The pores could be enlarged using 10 mM diamide, an SH residue oxidant that attacks surface proteins. We found that two strains showed signs of oxidative damage via changes in density and diameter of the surface pores. We found that the German strain was resistant to diamide induced oxidative damage, even when the concentration of the oxidant was increased to 50 mM. The normal pore size found on the cell walls of American strains had diameters of about 200nm. Under conditions of oxidative stress the diameters changed to 400nm.This method may prove to be a useful rapid screening process (45-60 min) to determine which strains are oxidative resistant, as well as being able to screen for groups of yeast that are sensitive to oxidative stress. This rapid screening tool may have direct applications in molecular biology (transference of the genes to inside of living cells) and biotechnology (biotransformations reactions to produce chiral synthons in organic chemistry.

Light microscopy and ultrastructure of the liver of Astyanax altiparanae Garutti and Britski, 2000 (Teleostei, Characidae).

Livers of thirty specimens of Astyanax altiparanae obtained from a commercial fish farm were subjected to light and transmission electron microscopy, in order to describe the hepatic parenchyma and the intrahepatic exocrine pancreatic tissue. Anatomically, the liver showed only three hepatic lobes. Histological analysis demonstrated that the hepatocytes were spread out as anastomotic cords, arranged in two cellular layers and surrounded by sinusoids. The intrahepatic exocrine pancreatic tissue exhibited an acinar arrangement and was diffused in the hepatic parenchyma. Ultrastructural analysis showed that the hepatocytes had a rounded nucleus and a rough endoplasmatic reticulum, with a parallel disposition to the nuclear membrane. The exocrine pancreatic cells showed secretion granules at the apical portion, and the rough endosplasmatic reticulum was concentrically distributed.

Quantitative fractography under light microscopy: A digital image processing approach

This work is an example of the improvement on quantitative fractography by means of digital image processing and light microscopy. Two techniques are presented to investigate the quantitative fracture behavior of Ti-4Al-4V heat-treated alloy specimens, under Charpy impact testing. The first technique is the Minkowski method for fractal dimension measurement from surface profiles, revealing the multifractal character of Ti-4Al-4V fracture. It was not observed a clear positive correlation of fractal values against Charpy energies for Ti-4Al-4V alloy specimens, due to their ductility, microstructural heterogeneities and the dynamic loading characteristics at region near the V-notch. The second technique provides an entire elevation map of fracture surface by extracting in-focus regions for each picture from a stack of images acquired at successive focus positions, then computing the surface roughness. Extended-focus reconstruction has been used to explain the behavior along fracture surface. Since these techniques are based on light microscopy, their inherent low cost is very interesting for failure investigations.

Digital Light Microscopy of Macroporous Ceramics Processed by Consolidation Casting with Mixtures of Native Starches

Porosity in starch consolidation casting technique is rightly related to original size and morphology of starch granules, leaving a pore structure after burning out. This work reports the results for the addition of different native potato and corn starch proportions in suspension,; with TiO(2) (rutile) powder. Gelling temperature have been defined after observation under light microscopy using a heating stage. Analysis of porous network and isolated pores have been clone from images of samples surfaces obtained by depth from focus reconstruction, revealing a qualitative correlation of pores characteristics and starches additions in suspensions, suggesting that the presence of isolated or interconnected pores can be handled by starches selection to control the amylopectin and amylose contents in slurries. Also, the analysis of porous fraction distribution shows no consistent pattern through specimens' volume according to starches in mixtures.