RESUMO
The interaction of the cytotoxic metals cadmium, zinc, and lead with pancreatic cells was studied by atomic force/lateral Force microscopy (AFM/LFM), an approach that provides both topographic (with nanometer scale lateral resolution) and chemical information on the membrane. Different morphological modifications of the overall cell shape and roughness took place as consequence of 100 muM metal-dependent treatment. Furthermore, after exposure to Cd(Cl(2)) and Zn(Cl(2)), but not Pb(Cl(2)), the LFM images revealed several areas of the cell's surface showing lateral friction contrasts that have been interpreted as marker of different alterations of the cell physiology induced by the metal loading. Thus, the coupling of LFM detection to topographic AFM characterization allows to distinguish, through a nondestructive and surface characterising approach, between different metal-induced cytotoxic effects on cells. In this framework, the role of the LFM as an important tool to discriminate between different alteration of a biological system has to be highlighted.
Assuntos
Membrana Celular/efeitos dos fármacos , Células Secretoras de Insulina/citologia , Células Secretoras de Insulina/efeitos dos fármacos , Metais Pesados/toxicidade , Microscopia de Força Atômica/métodos , Apoptose/efeitos dos fármacos , Cloreto de Cádmio/metabolismo , Cloreto de Cádmio/toxicidade , Membrana Celular/ultraestrutura , Forma Celular/efeitos dos fármacos , Cloretos/metabolismo , Cloretos/toxicidade , Humanos , Chumbo/metabolismo , Chumbo/toxicidade , Metais Pesados/metabolismo , Compostos de Zinco/metabolismo , Compostos de Zinco/toxicidadeRESUMO
In this paper we present chemically highly resolved images obtained with Scanning Near-field Optical Microscopy (SNOM) coupled with an Infrared (IR) Free Electron Laser (FEL) at Vanderbilt University, Nashville, USA. Main principles governing SNOM imaging as well as essential components of the experimental setup are described. Chemically resolved images showing the distribution of different phases within the boron-nitride films are presented. Universal character of the experiment and its huge potential applications in biophysics and medical sciences domain are illustrated with highly resolved SNOM images of pancreatic cells.
Assuntos
Microscopia de Força Atômica/métodos , Espectrofotometria Infravermelho/métodos , Animais , Linhagem Celular , Nanotecnologia , Pâncreas/ultraestrutura , RatosRESUMO
Human skin cell culture (HaCaT) that has been exposed to an AC magnetic field undergoes detectable changes in its biochemical properties and shapes. Such changes were observed by infrared wavelength-selective scanning near-field optical microscopy with a resolution of 80-100 nm. We specifically investigated the changes in the distribution of the inner chemical functional groups and in the cell morphology induced by a 24 h exposure to a 1 mT (rms), 50 Hz sinusoidal magnetic field in a temperature regulated solenoid. These results further accentuate the crucial questions, raised by several recent studies, about the impact of low-frequency electromagnetic field on human cells.
Assuntos
Campos Eletromagnéticos , Células Epiteliais/efeitos da radiação , Radiação não Ionizante , Pele/efeitos da radiação , Adesão Celular , Técnicas de Cultura de Células , Linhagem Celular , Células Epiteliais/citologia , Desenho de Equipamento , Humanos , Raios Infravermelhos , Microscopia/métodos , Pele/citologia , TemperaturaRESUMO
The infrared (IR) absorption of a biological system can potentially report on fundamentally important microchemical properties. For example, molecular IR profiles are known to change during increases in metabolic flux, protein phosphorylation, or proteolytic cleavage. However, practical implementation of intracellular IR imaging has been problematic because the diffraction limit of conventional infrared microscopy results in low spatial resolution. We have overcome this limitation by using an IR spectroscopic version of scanning near-field optical microscopy (SNOM), in conjunction with a tunable free-electron laser source. The results presented here clearly reveal different chemical constituents in thin films and biological cells. The space distribution of specific chemical species was obtained by taking SNOM images at IR wavelengths (lambda) corresponding to stretch absorption bands of common biochemical bonds, such as the amide bond. In our SNOM implementation, this chemical sensitivity is combined with a lateral resolution of 0.1 micro m ( approximately lambda/70), well below the diffraction limit of standard infrared microscopy. The potential applications of this approach touch virtually every aspect of the life sciences and medical research, as well as problems in materials science, chemistry, physics, and environmental research.