Characterizing wear processes on orthopaedic materials using scanning probe microscopy

The operational lifetime of hip replacement prostheses can be severely limited due to the occurrence of excessive wear at the load-bearing interfaces. The aim of this study was to investigate how the surface topography of articulating counterfaces evolves over the duration of a laboratory wear run. It was observed that modular stainless steel femoral heads wearing against ultrahigh molecular weight polyethylene (UHMWPE) can themselves be subject to wearing. A comparison with retrieved in vivo-aged femoral heads shows many topographical similarities: in a qualitative sense, scratching and pitting are evident on laboratory and in vivo-worn femoral heads; quantitatively, roughness comparisons between the new and worn devices are seen to increase typically by a factor of 4 after laboratory wearing. The observations suggest that a particular wear mode, namely third-body wear, is responsible for the increased roughness. It is conjectured that third bodies might arise through surface fatigue wear on the metal counterface, Wear debris is also observed to have been generated from the polymer surface, creating rounded debris with sizes predominantly in the range 0.4-0.8 microns: dimensions that are comparable to values previously reported for in vivo generated debris.

Light and electron microscopy of microstructure and fractography of an ultrahigh-strength martensitic steel

Microstructure, tensile properties and fractography have been examined in the oil-quenched samples of a low-alloy ultrahigh strength 4340 steel. Intergranular fracture was revealed to locate at the fracture origin. However, neither the quenched Charpy V-notched impact samples nor the tempered tensile samples showed such intergranular fracture behavior. The effects of loading rate and precipitation are discussed.

Manipulating plasmon modes in tip-enhanced Raman microscopy

The influence of polarization on the plasmon modes excited in tip-enhanced near-field optical microscopy have been investigated using the Finite Difference Time Domain Method. Analysis of the calculated results have laid particular emphasis on the ability to align local field enhancements with the orientation of molecules in order to optimize Raman signals, with particular relevance to recent experimental work on carbon nanotubes.

Scaled Heavy-Ball Acceleration of the Richardson-Lucy Algorithm for 3D Microscopy Image Restoration

The Richardson–Lucy algorithm is one of the most important in image deconvolution. However, a drawback is its slow convergence. A significant acceleration was obtained using the technique proposed by Biggs and Andrews (BA), which is implemented in the deconvlucy function of the image processing MATLAB toolbox. The BA method was developed heuristically with no proof of convergence. In this paper, we introduce the heavy-ball (H-B) method for Poisson data optimization and extend it to a scaled H-B method, which includes the BA method as a special case. The method has a proof of the convergence rateof O(K^2), where k is the number of iterations. We demonstrate the superior convergence performance, by a speedup factor off ive, of the scaled H-B method on both synthetic and real 3D images.

Progress in the use of scanning electron microscopy (SEM) applied to the taxonomy of the genus Bursaphelenchus (Nematoda: Parasitaphelenchidae)

The scanning electron microscope (SEM) has been a major tool in detailed morphological observations of plant parasitic nematodes during the last 30 years, efficiently complementing light microscopical (LM) studies. Nematodes are extremely difficult to observe and characterize due to their small size (aprox. 1 mm long) and paucity of morphological characters, so detailed surface observations of several organs and nematode regions are of the highest value. Among plant parasitic nematodes, one of the most devastating species is the “pinewood nematode” (PWN), Bursaphelenchus xylophilus, which has been a major problem for forest species, and in particular pines, in Asia (Japan, China, Korea) and has been recently detected in the European Union (Portugal). B. xylophilus belongs to a closely related, morphologically similar group of species, within the genus Bursaphelenchus, and designated by the “xylophilus group”. SEM has become a crucial tool in observing several genital characters of males and females, such as male genital papillae, male copulatory spicules, female vulval flap and female genital papillae.s In this presentation, we will show how SEM has been utilized to observe and characterize the shape of the vulval flap, the presence/ absence of papillae near the flap, and confirm the presence and the arrangement of the male genital papillae. LM is also used in this work to show its value as a complementary tool to SEM, in both genital characteristics and other, general, characters of the genus Bursaphelenchus, such as the male bursa and cephalic region.

Piezoresponse force microscopy of ferroelectric relaxors

Nesta tese, ferroeléctricos relaxor (I dont know uf the order is correct) de base Pb das familias (Pb,La)(Zr,Ti)O3 (PLZT), Pb(Mg1/3,Nb2/3)O3-PbTiO3 (PMN-PT), Pb(Zn1/3,Nb2/3)O3-PbTiO3 (PZN-PT) foram investigados e analisados. As propriedades ferroeléctricas e dieléctricas das amostras foram estudadas por métodos convencionais de macro e localmente por microscopia de força piezoeléctrica (PFM). Nos cerâmicos PLZT 9.75/65/35 o contraste da PFM à escala nanometrica _ foi investigado em função do tamanho e orientação dos grãos. Apurou-se que a intensidade do sinal piezoeléctrico das nanoestruturas diminui com o aumento da temperatura e desaparece a 490 K (La mol. 8%) e 420 K (9,5%). Os ciclos de histerese locais foram obtidos em função da temperatura. A evolução dos parâmetros macroscópicos e locais com a temperatura de superfície sugere um forte efeito de superfície nas transições de fase ferroeléctricas do material investigado. A rugosidade da parede de domínio é determinada por PFM para a estrutura de domínio natural existente neste ferroeléctrico policristalino. Além disso, os domínios ferroeléctricos artificiais foram criados pela aplicação de pulsos eléctricos à ponta do condutor PFM e o tamanho de domínio in-plane foi medido em função da duração do pulso. Todas estas experiências levaram à conclusão de que a parede de domínio em relaxors do tipo PZT é quase uma interface unidimensional. O mecanismo de contraste na superfície de relaxors do tipo PLZT é medido por PFMAs estruturas de domínio versus evolução da profundidade foram estudadas em cristais PZN-4,5%PT, com diferentes orientações através da PFM. Padrões de domínio irregulares com tamanhos típicos de 20-100 nm foram observados nas superfícies com orientação <001> das amostras unpoled?. Pelo contrário, os cortes de cristal <111> exibem domínios regulares de tamanho mícron normal, com os limites do domínio orientados ao longo dos planos cristalográficos permitidos. A existência de nanodomínios em cristais com orientação <001> está provisoriamente (wrong Word) atribuída à natureza relaxor de PZN-PT, onde pequenos grupos polares podem formar-se em coindições de zero-field-cooling (ZFC). Estes nanodomínios são considerados como os núcleos do estado de polarização oposta e podem ser responsáveis pelo menor campo coercitivo para este corte de cristal em particular. No entanto, a histerese local piezoelétrica realizada pelo PFM à escala nanométrica indica uma mudança de comportamento de PZN-PT semelhante para ambas as orientações cristalográficas investigadas. A evolução das estruturas de domínio com polimento abaixo da superfície do cristal foi investigada. O domínio de ramificações e os efeitos de polarização de triagem após o polimento e as medições de temperatura têm sido estudados pela PFM e pela análise SEM. Além disso, verificou-se que a intensidade do sinal piezoeléctrico a partir das estruturas de nanodomínio diminui com o aumento da temperatura, acabando por desaparecer aos 430 K (orientaçáo <111>) e 470 K (orientação <100>). Esta diferença de temperatura nas transições de fase local em cristais de diferentes orientações é explicada pelo forte efeito de superfície na transição da fase ferroelétrica em relaxors.A comutação da polarização em relaxor ergódico e nas fases ferroeléctricas do sistema PMN-PT foram realizadas pela combinação de três métodos, Microscopia de Força Piezoeléctrica, medição de um único ponto de relaxamento eletromecânico e por ultimo mapeamento de espectroscopia de tensão. A dependência do comportamento do relaxamento na amplitude e tempo da tensão de pulso foi encontrada para seguir um comportamento logarítmico universal com uma inclinação quase constante. Este comportamento é indicativo da progressiva população dos estados de relaxamento lento, ao contrário de uma relaxação linear na presença de uma ampla distribuição do tempo de relaxamento. O papel do comportamento de relaxamento, da não-linearidade ferroeléctrica e da heterogeneidade espacial do campo na ponta da sonda de AFM sobre o comportamento do ciclo de histerese é analisada em detalhe. Os ciclos de histerese para ergódica PMN- 10%PT são mostrados como cineticamente limitados, enquanto que no PMN, com maior teor de PT, são observados verdadeiros ciclos de histerese ferroeléctrica com viés de baixa nucleação.

Identification of bacteria associated with Dinoflagellates (Dinophyceae) Alexandrium spp. using tyramide signal amplification fluorescent in situ hybridization and confocal microscopy

In the marine environment, phytoplankton and bacterioplankton can be physically associated. Such association has recently been hypothesized to be involved in the toxicity of the dinoflagellate genus Alexandrium. However, the methods, which have been used so far to identify, localize, and quantify bacteria associated with phytoplankton, are either destructive, time consuming, or lack precision. In the present study we combined tyramide signal amplification–fluorescent in situ hybridization (TSA-FISH) with confocal microscopy to determine the physical association of dinoflagellate cells with bacteria. Dinoflagellate attached microflora was successfully identified with TSA-FISH, whereas FISH using monolabeled probes failed to detect bacteria, because of the dinoflagellate autofluorescence. Bacteria attached to entire dinoflagellates were further localized and distinguished from those attached to empty theca, by using calcofluor and DAPI, two fluorochromes that stain dinoflagellate theca and DNA, respectively. The contribution of specific bacterial taxa of attached microflora was assessed by double hybridization. Endocytoplasmic and endonuclear bacteria were successfully identified in the nonthecate dinoflagellate Gyrodinium instriatum. In contrast, intracellular bacteria were not observed in either toxic or nontoxic strains of Alexandrium spp. Finally, the method was successfully tested on natural phytoplankton assemblages, suggesting that this combination of techniques could prove a useful tool for the simultaneous identification, localization, and quantification of bacteria physically associated with dinoflagellates and more generally with phytoplankton.

Taking nanomedicine teaching into practice with atomic force microscopy and force spectroscopy

© 2015 The American Physiological Society

Comparative study of human erythrocytes by digital holographic microscopy, confocal microscopy, and impedance volume analyzer.

Red blood cell (RBC) parameters such as morphology, volume, refractive index, and hemoglobin content are of great importance for diagnostic purposes. Existing approaches require complicated calibration procedures and robust cell perturbation. As a result, reference values for normal RBC differ depending on the method used. We present a way for measuring parameters of intact individual RBCs by using digital holographic microscopy (DHM), a new interferometric and label-free technique with nanometric axial sensitivity. The results are compared with values achieved by conventional techniques for RBC of the same donor and previously published figures. A DHM equipped with a laser diode (lambda = 663 nm) was used to record holograms in an off-axis geometry. Measurements of both RBC refractive indices and volumes were achieved via monitoring the quantitative phase map of RBC by means of a sequential perfusion of two isotonic solutions with different refractive indices obtained by the use of Nycodenz (decoupling procedure). Volume of RBCs labeled by membrane dye Dil was analyzed by confocal microscopy. The mean cell volume (MCV), red blood cell distribution width (RDW), and mean cell hemoglobin concentration (MCHC) were also measured with an impedance volume analyzer. DHM yielded RBC refractive index n = 1.418 +/- 0.012, volume 83 +/- 14 fl, MCH = 29.9 pg, and MCHC 362 +/- 40 g/l. Erythrocyte MCV, MCH, and MCHC achieved by an impedance volume analyzer were 82 fl, 28.6 pg, and 349 g/l, respectively. Confocal microscopy yielded 91 +/- 17 fl for RBC volume. In conclusion, DHM in combination with a decoupling procedure allows measuring noninvasively volume, refractive index, and hemoglobin content of single-living RBCs with a high accuracy.

Simultaneous cell morphometry and refractive index measurement with dual-wavelength digital holographic microscopy and dye-enhanced dispersion of perfusion medium.

Digital holographic microscopy (DHM) allows optical-path-difference (OPD) measurements with nanometric accuracy. OPD induced by transparent cells depends on both the refractive index (RI) of cells and their morphology. This Letter presents a dual-wavelength DHM that allows us to separately measure both the RI and the cellular thickness by exploiting an enhanced dispersion of the perfusion medium achieved by the utilization of an extracellular dye. The two wavelengths are chosen in the vicinity of the absorption peak of the dye, where the absorption is accompanied by a significant variation of the RI as a function of the wavelength.