Experimental evaluation of a non-linear coded excitation method for contrast imaging

Previously, we have shown that for a single bubble, using chirps as the excitation signal improves both the linear and the non-linear response. Computer simulations of randomly distributed contrast agent bubbles show an increase of 10-13 dB in response when comparing pulse excitations with chirp excitations that have equal bandwidths and peak amplitudes. Second harmonic compression of simulated bubble echoes with chirp excitation shows low side-lobe levels and limited loss of axial resolution when compared to pulse excitation. Experimental results from water tank measurements with SonoVue contrast agent are in agreement with computer simulations showing increased signal-to-noise ratio and an increase of approximately 12 dB at the second harmonic when comparing pulse and chirp excitation.     

High-resolution confocal microscopy by saturated excitation of fluorescence

We demonstrate the use of saturated excitation in confocal fluorescence microscopy to improve the spatial resolution. In the proposed technique, we modulate the excitation intensity temporally and detect the harmonic modulation of the fluorescence signal which is caused by the saturated excitation in the center of the laser focus. Theoretical and experimental investigations show that the demodulated fluorescence signal is nonlinearly proportional to the excitation intensity and contributes to improve the spatial resolution in three dimensions beyond the diffraction limit of light.     

Synchrotron scanning photoemission microscopy of homogeneous and heterogeneous metal sulfide minerals

Scanning photoemission microscopy (SPEM) has been applied to the investigation of homogeneous and heterogeneous metal sulfide mineral surfaces. Three mineral samples were investigated: homogeneous chalcopyrite, heterogeneous chalcopyrite with bornite, and heterogeneous chalcopyrite with pyrite. Sulfur, copper and iron SPEM images, i.e. surface‐selective elemental maps with high spatial resolution acquired using the signal from the S 2p and Cu and Fe 3p photoemission peaks, were obtained for the surfaces after exposure to different oxidation conditions (either exposed to air or oxidized in pH 9 solution), in addition to high‐resolution photoemission spectra from individual pixel areas of the images. Investigation of the homogeneous chalcopyrite sample allowed for the identification of step edges using the topography SPEM image, and high‐resolution S 2p spectra acquired from the different parts of the sample image revealed a similar rate of surface oxidation from solution exposure for both step edge and a nearby terrace site. SPEM was able to successfully distinguish between chalcopyrite and bornite on the heterogeneous sample containing both minerals, based upon sulfur imaging. The high‐resolution S 2p spectra acquired from the two regions highlighted the faster air oxidation of the bornite relative to the chalcopyrite. Differentiation between chalcopyrite and pyrite based upon contrast in SPEM images was not successful, owing to either the poor photoionization cross section of the Cu and Fe 3p electrons or issues with rough fracture of the composite surface. In spite of this, high‐resolution S 2p spectra from each mineral phase were successfully obtained using a step‐scan approach.     

Near-field scanning optical microscopy of single molecules by femtosecond two-photon excitation

We present a demonstration of near-field scanning optical microscopy of single molecules based on ultrafast two-photon-induced fluorescence. Measurements were performed by use of 100-fs pulses at 800 nm from a Ti:sapphire laser to excite the two-photon transition in Rhodamine B molecules. Although near-field probes are normally metal coated to achieve superresolution, we used uncoated tips to achieve sufficiently high optical powers to generate acceptable fluorescence emission rates. Images of single molecules demonstrate a resolution of ~175nm(< lambda/4) on a topographically smooth surface, which surpasses the apparent lambda/2 resolution limit for uncoated tips operating in the linear response regime.     

4Pi microscopy with linear fluorescence excitation

Practical 4Pi microscopy has so far exclusively relied on multiphoton excitation of fluorescence, because the nonlinear suppression of contributions from higher-order sidelobes was mandatory for unambiguous axial superresolution. We show that novel lenses of 74 degrees semiaperture angle enable biological 4Pi microscopy with regular one-photon fluorescence excitation, thus increasing the signal and reducing system complexity and cost. An axial resolution of 95 nm, corresponding to a more than fourfold improvement over confocal microscopy, is verified in the imaging of microtubules in mammalian cells.     

Hyperfine level-crossing spectroscopy of NH2 with dye-laser excitation

Hyperfine structure of a number of levels of the (0, 9, 0) and (0, 10, 0) vibrational states of the à ²A₁ state of NH₂ has been measured using the technique of magnetic level-crossing. A CW dye-laser served as an excitation source. The resolution achieved is ～ 3 MHz. The analyses have been aided by calculations of the relative fields for the level-crossings in terms of the parameters of a model hamiltonian, together with their relative intensities for linear and non-linear excitation. These calculations are presented for I = ½ and 1 for J = 1 ½, 2 ½, 3 ½, and 4 ½.

Hyperfine parameters for the (0, 10, 0) vibrational state are in good agreement with the one set of literature values, obtained using the independent method of intermodulated fluorescence.

We observe an apparent drop of 30 per cent in the Fermi contact parameter between the (0, 10, 0) and (0, 9, 0) levels. However, it is also found that the hyperfine splittings within the (0, 9, 0) state are not completely in accord with the simple model. These observations are briefly discussed in terms of the Renner effect and the s-p hybrid nature of the molecular orbital occupied by the unpaired electron.     

Tailored sinc pulses for uniform excitation and artifact-free radio frequency time-domain EPR imaging

A method to generate shaped radiofrequency pulses for uniform excitation of electron spins in time-domain radio frequency (RF) electron paramagnetic resonance (EPR) imaging is presented. A commercial waveform generator was integrated with the transmit arm of the existing time-domain RF-EPR spectrometer to generate tailored excitation pulses with sub-nano second resolution for excitation with a 90 degrees flip-angle. A truncated sinc [sin(x)/x] pulse, tailored to compensate for the Q-profile (RF frequency response) of the resonator, was shown to yield images from phantom objects as well as in vivo images, with minimal distortion. These studies point to the advantages in using shaped sinc pulses to achieve improved uniform excitation over a relatively wide bandwidth region in time-domain RF-EPR imaging (RF-FT-EPRI).     

Three-dimensional resolution enhancement in fluorescence microscopy by harmonic excitation

A method for increasing lateral as well as axial resolution in fluorescence microscopy is presented. A passband with a high cutoff frequency throughout reciprocal space can be achieved by illumination of the object with spatially harmonic excitation patterns generated by the interference of two collimated laser beams. Theoretical calculations show an almost isotropic point-spread function with a FWHM near 100 nm.     

饱和激发情况下的荧光关联谱测量

研究了激发光增强时其对荧光关联谱(FCS)测量的影响. 强激发光产生的饱和激发会改 变激光诱导荧光的空间分布函数,从而影响FCS的测量结果. 首先根据饱和吸收的物理模型,推导出强激发光情况下有效探测区域变化的定量公式,获得了FCS测量所得到的粒子数和扩散时间与饱和激发光强和激光光强的函数关系,并用于消除饱和吸收造成的系统误差.采用Monte-Carlo模拟方法和实验,对理论分析结果进行了验证. 这将有助于完善FCS分析方法的理论模型,为高激发光强度下的FCS探测提供依据. 关键词： 荧光关联谱 饱和激发 Monte-Carlo模拟     

Chirp excitation of ultrasonic guided waves

Most ultrasonic guided wave methods require tone burst excitations to achieve some degree of mode purity while maintaining temporal resolution. In addition, it is often desirable to acquire data using multiple frequencies, particularly during method development when the best frequency for a specific application is not known. However, this process is inconvenient and time-consuming, particularly if extensive signal averaging at each excitation frequency is required to achieve a satisfactory signal-to-noise ratio. Both acquisition time and data storage requirements may be prohibitive if responses from many narrowband tone burst excitations are measured. Here chirp excitations are utilized to address the need to both test at multiple frequencies and achieve a high signal-to-noise ratio to minimize acquisition time. A broadband chirp is used to acquire data at a wide range of frequencies, and deconvolution is applied to extract multiple narrowband responses. After optimizing the frequency and duration of the desired tone burst excitation, a long-time narrowband chirp is used as the actual excitation, and the desired tone burst response is similarly extracted during post-processing. Results are shown that demonstrate the efficacy of both broadband and narrowband chirp excitations.     

Reducing SAR in parallel excitation using variable-density spirals: a simulation-based study

Parallel excitation using multiple transmit channels has emerged as an effective method to shorten multidimensional spatially selective radiofrequency (RF) pulses, which have a number of important applications, including B1 field inhomogeneity correction in high-field MRI. The specific absorption rate (SAR) is a primary concern in high-field MRI, where wavelength effects can lead to local peaks in SAR. In parallel excitation, the subjects are exposed to RF pulses from multiple coils, which makes the SAR problem more complex to analyze, yet potentially enables greater freedom in designing RF pulses with lower SAR. Parallel-excitation techniques typically employ either Cartesian or constant-density (CD) spiral trajectories. In this article, variable-density (VD) spiral trajectories are explored as a means for SAR reduction in parallel-excitation pulse design. Numerical simulations were conducted to study the effects of CD and VD spirals on parallel excitation. Specifically, the electromagnetic fields of a four-channel transmit head coil with a three-dimensional head model at 4.7 T were simulated using a finite-difference time domain method. The parallel RF pulses were designed and the resulting excitation patterns were generated using a Bloch simulator. The SAR distributions due to CD and VD spirals were evaluated quantitatively. The simulation results show that, for the same pulse duration, parallel excitation with VD spirals can achieve a lower SAR compared to CD spirals for parallel excitation. VD spirals also resulted in reduced artifact power in the excitation patterns. This gain came with slight, but noticeable, degrading of the spatial resolution of the resulting excitation patterns.     

双光子技术在三维成像和三维存储技术中的应用

对双光子过程的空间分辨能力进行了理论分析。阐明了双光子技术在三维成像和三维存储中的独特优势。着重介绍了双光子技术与扫描共焦显微术、近场显微术相结合进行三维成像 ,以及一种多焦点多光子显微术和用连续光源激发的双光子三维成像技术的研究和进展情况。对建立在共焦显微镜基础之上的双光子三维光存储和微细加工方面的研究也作了回顾与展望     

Spatially distributed two-photon excitation fluorescence in scattering media: Experiments and time-resolved Monte Carlo simulations

We report on measurements and Monte Carlo analysis of the spatial distribution of light in two-photon excitation fluorescence (TPEF) microscopy operating through scattering media. The axial profile of TPEF produced by a high numerical aperture microscope objective focusing through a tissue-like optical phantom at depths more than five photon mean-scattering lengths is studied. The measured profile is quantitatively interpreted with time-resolved Monte Carlo simulations that take into account the spatio-temporal distribution of the photons within the turbid medium under a femtosecond excitation regime. It is shown that considering only the ballistic photons leads to an under-estimation of the actual out-of-focus distribution of the fluorescence, whereas it is over-estimated when the ballistic and scattered photons are treated alike. Comparison of the in-focus signal to the overall out-of-focus background clearly pointed out that the signal-to-background ratio of TPEF microscopy images benefits from a shortening of the excitation pulses. Moreover, in homogeneously labeled specimen, the background is shown to overcome the signal at imaging depths of about five to six photon mean-scattering paths.     

Far-field fluorescence microscopy with repetitive excitation

We introduce the concept of repeatedly exciting an excited state of a photostable fluorescent entity to generate a nonlinear fluorescence signal which is solely based on the linear susceptibility of the molecule. The excitation cycle between the fluorescent state and another state prolongs the average lifetime of , with emphasis on those molecules that are in the center of the focus. The photons emitted by the long-lived molecules in the center are recorded by a temporal filter and constitute fluorescence that depends nonlinearly on the excitation intensity. Theoretical analysis reveals that this concept can provide three-dimensional imaging and improve the spatial resolution in far-field fluorescence microscopy. We show that despite the presence of diffraction the effective focal waist can in principle be narrowed down to the molecular scale at the expense of signal. Received: 3 December 1998     

Performance improvement of magneto-acousto-electrical tomography for biological tissues with sinusoid-Barker coded excitation

By combining magnetics, acoustics and electrics, the magneto-acoustic-electrical tomography(MAET) proves to possess the capability of differentiating electrical impedance variation and thus improving the spatial resolution. However,the signal-to-noise ratio(SNR) of the collected MAET signal is still unsatisfactory for biological tissues with low-level electrical conductivity. In this study, the formula of MAET measurement with sinusoid-Barker coded excitation is derived and simplified for a planar piston transducer. Numerical simulations are conducted for a four-layered gel phantom with the 13-bit sinusoid-Barker coded excitation, and the performances of wave packet recovery with side-lobe suppression are improved by using the mismatched compression filter, which is also demonstrated by experimentally measuring a three-layered gel phantom. It is demonstrated that comparing with the single-cycle sinusoidal excitation, the amplitude of the driving signal can be reduced greatly with an SNR enhancement of 10 dB using the 13-bit sinusoid-Barker coded excitation. The amplitude and polarity of the wave packet filtered from the collected MAET signal can be used to achieve the conductivity derivative at the tissue boundary. In this study, we apply the sinusoid-Barker coded modulation method and the mismatched suppression scheme to MAET measurement to ensure the safety for biological tissues with improved SNR and spatial resolution, and suggest the potential applications in biomedical imaging.     

Characterization and analysis of coded excitation ultrasound parameters for rock properties

Coded excitation technology(CET) can effectively enhance the penetration and resolution of ultrasonic testing.To analyze the influence of rock properties on pulse compression performance(PCP) of coded excitation signals(CES),a numerical simulation,and an ultrasonic experiment on different rock samples are performed;and the detection ability of several CESs are also investigated and compared.The results of experiments showed that the loss of the signal-to-noise ratio(SNR) of Barker coded signal with tapered linear frequency modulated carrier(BTLFM) is always less than Barker coded signal with sine carrier(BS),while the resolution loss of BTLFM is lower than tapered linear frequency modulated signal(TLFM).In sum,the results not only verifiy the effectiveness of CET,but also provide a basis for the parameter settings of coded signals used in rock ultrasonic testing.     

Local 2PPE-yield enhancement in a defined periodic silver nanodisk array

Well-prepared periodic arrays of silver nanoparticles are investigated by means of linear and non-linear photoemission electron microscopy. The structures show homogeneous photoemission for UV excitation in the linear photoemission regime whereas striking inhomogeneities are mapped in the case of the nonlinear (2 photon) excitation using ultrashort 400 nm laser pulses. A detailed analysis enables to assign these inhomogeneities to defect induced electron momentum transfer processes only effective for the 2 photon excitation process. We propose this mechanism to be of relevance for the appearance of so-called hot spots in nonlinear photoemission as identified in other 2PPE studies in the past. Furthermore, the complementarity between all-optical studies and nonlinear photoemission studies of localized surface plasmons in nanoparticles is discussed.     

Biomolecular imaging based on far-red fluorescent protein with a high two-photon excitation action cross section

Received October 14, 2005; revised January 7, 2006; accepted January 9, 2006; posted January 12, 2006 (Doc. ID 65391) The two-photon excitation action cross section of Hc-Red fluorescent proteins (Hc-RFPs) is measured and found to be of the same order as that of enhanced green fluorescent proteins. With a 618 nm emission wavelength in the far-red region and with an excitation wavelength around 1200 nm, Hc-RPF-based two-photon fluorescence microscopy (2PFM) can offer deep penetration capability inside live samples and is ideal for in vivo gene expression study and biomolecular imaging in live objects. In vivo 2PFM of the developing heart deep inside a transgenic zebrafish embryo tagged by Hc-RFP is also successfully demonstrated.     

Multi-resolution technique integrated with smoothed particle element method (SPEM) for modeling fluid-structure interaction problems with free surfaces

Free-surface flows, especially those associated with fluid-structure interactions(FSIs), pose challenging problems in numerical simulations. The authors of this work recently developed a smoothed particle element method(SPEM) to simulate FSIs. In this method, both the fluid and solid regions are initially modeled using a smoothed finite element method(S-FEM) in a Lagrangian frame, whereas the fluid regions undergoing large deformations are adaptively converted into particles and modeled with an improved smoothed particle hydrodynamics(SPH) method. This approach greatly improves computational accuracy and efficiency because of the advantages of the S-FEM in efficiently treating solid/fluid regions showing small deformations and the SPH method in effectively modeling moving interfaces. In this work, we further enhance the efficiency of the SPEM while effectively capturing local fluid information by introducing a multi-resolution technique to the SPEM and developing an effective approach to treat multi-resolution element-particle interfaces. Various numerical examples demonstrate that the multiresolution SPEM can significantly reduce the computational cost relative to the original version with a constant resolution.Moreover, the novel approach is effective in modeling various incompressible flow problems involving FSIs.     

On the influence of backscattered electrons on the lateral resolution in scanning auger microscopy

A definition of edge resolution is proposed, which is adapted to the pecularities of scanning Auger microscopy. Based on recent monte-Carlo computer simulations for scanning electron microscopy, the influence of backscattered electrons on the edge resolution is estimated for low-Z (Al) and high-Z materials (Au). The resolution is found to be of the order of 100nm and to be nearly independent of the atomic number of the sample.