Second harmonic generation and multiphoton microscopic detection of collagen without the need for species specific antibodies

High-resolution, high-contrast, three-dimensional images of live cell and tissue architecture can be obtained using second harmonic generation (SHG), which comprises non-absorptive frequency changes in an excitation laser line. SHG does not require any exogenous antibody or fluorophore labeling, and can generate images from unstained sections of several key endogenous biomolecules, in a wide variety of species and from different types of processed tissue. Here, we examined normal control human skin sections and human burn scar tissues using SHG on a multi-photon microscope (MPM). Examination and comparison of normal human skin and burn scar tissue demonstrated a clear arrangement of fibers in the dermis, similar to dermal collagen fiber signals. Fluorescence-staining confirmed the MPM-SHG collagen colocalization with antibody staining for dermal collagen type-I but not fibronectin or elastin. Furthermore, we were able to detect collagen MPM-SHG signal in human frozen sections as well as in unstained paraffin embedded tissue sections that were then compared with hematoxylin and eosin staining in the identical sections. This same approach was also successful in localizing collagen in porcine and ovine skin samples, and may be particularly important when species-specific antibodies may not be available. Collectively, our results demonstrate that MPM SHG-detection is a useful tool for high resolution examination of collagen architecture in both normal and wounded human, porcine and ovine dermal tissue.

LED lighting design strategies to enhance window appearance and increase energy savings in daylit office spaces

Vertical windows are the most common and simplest method to introduce daylight to interior spaces of office buildings, while also providing a view and connection to the outside. However, high contrast ratios between windows and surrounding surfaces can cause visual discomfort for occupants and can negatively influence their health and productivity. Consequently, building occupants may try to adapt their working environment through closing blinds and turning on lights in order to improve indoor visual comfort. Such interventions defeat the purpose of daylight harvesting systems and can increase the forecast electric lighting consumption in buildings that include such systems. A simple strategy to prevent these problematic consequences is to reduce the luminance contrasts presented by the window wall by increasing the luminance of areas surrounding the window through the sparing use of energy-efficient supplementary lighting, such light emitting diodes (LEDs). This paper presents the result of a pilot study in typical office in Brisbane, Australia that tests the effectiveness of a supplementary LED lighting system. The study shows an improvement in the appraisal of the visual environment is achieved using the supplementary system, along with up to 88% reductions in luminance contrast at the window wall. Also observed is a 36% reduction in the likelihood of user interventions that would increase energy usage. These results are used as the basis of an annual energy simulation of the test office and indicate that supplementary systems could be used to save energy beyond what is typically realised in side lit office spaces.

Pilot study: Effect of age on visual acuity with defocus and astigmatism

PURPOSE: To investigate how distance visual acuity in the presence of defocus and astigmatism is affected by age and whether aberration properties of young and older eyes can explain any differences. METHODS: Participants were 12 young adults (mean [±SD] age, 23 [±2] years) and 10 older adults (mean [±SD] age, 57 [±4] years). Cyclopleged right eyes were used with 4-mm effective pupil sizes. Thirteen blur conditions were used by adding five spherical lens conditions (-1.00 diopters [D], -0.50 D, plano/0.00 D, +0.50 D, and +1.00 D) and adding two cross-cylindrical lenses (+0.50 DS/-1.00 DC and +1.00 D/-2.00 DC, or 0.50 D and 1.00 D astigmatism) at four negative cylinder axes (45, 90, 135, and 180 degrees). Targets were single lines of high-contrast letters based on the Bailey-Lovie chart. Successively smaller lines were read until a participant could no longer read any of the letters correctly. Aberrations were measured with a COAS-HD Hartmann-Shack aberrometer. RESULTS: There were no significant differences between the two age groups. We estimated that 70 to 80 participants per group would be needed to show significant effects of the trend of greater visual acuity loss for the young group. Visual acuity loss for astigmatism was twice that for defocus of the same magnitude of blur strength (0.33 logMAR [logarithm of the minimum angle of resolution]/D compared with 0.18 logMAR/D), contrary to the geometric prediction of similar loss. CONCLUSIONS: Any age-related differences in visual acuity in the presence of defocus and astigmatism were swamped by interparticipant variation.

Integrating LED lighting design strategies with side daylighting systems to improve interior lighting design of office buildings

A high contrast ratio between windows and surrounding walls may lead to office workers visual discomfort that could negatively affect their satisfaction and productivity. Consequently, occupants may try to adapt their working environment by closing blinds and/ or turning on the lights to enhance indoor visual comfort, which can reduce predicted energy savings. The hypothesis of this study is that reducing luminance contrast ratio on the window wall will improve window appearance which potentially will reduce visual discomfort and decrease workers interventions. Thus, this PhD research proposes a simple strategy to diminish the luminance contrast on the window wall by increasing the luminance of the areas surrounding the windows using supplementary light emitting diode (LED) systems. To test the hypothesis, this investigation will involve three experiments in different office layouts with various window types and orientations in Brisbane, Australia. It will assess user preferences for different luminance patterns in windowed offices featuring flexible, lowpower LED lighting installations that allows multiple lighting design options on the window wall. Detailed luminance and illuminance measures will be used to match quantitative lighting design assessment to user preferences.

Pattern visual evoked potentials in dyslexic versus normal children

Purpose: Presence of neurophysiological abnormalities in dyslexia has been a conflicting issue. This study was performed to evaluate the role of sensory visual deficits in the pathogenesis of dyslexia. Methods: Pattern visual evoked potentials (PVEP) were recorded in 72 children including 36 children with dyslexia and 36 children without dyslexia (controls) who were matched for age, sex and intelligence. Two check sizes of 15 and 60 min of arc were used with temporal frequencies of 1.5 Hz for transient and 6 Hz for steady‑state methods. Results: Mean latency and amplitude values for 15 min arc and 60 min arc check sizes using steady state and transient methods showed no significant difference between the two study groups (P values: 0.139/0.481/0.356/0.062).Furthermore, no significant difference was observed between two methods of PVEPs in dyslexic and normal children using 60min arc with high contrast(Pvalues: 0.116, 0.402, 0.343 and 0.106). Conclusion: The sensitivity of PVEP has high validity to detect visual deficits in children with dyslexic problem. However, no significant difference was found between dyslexia and normal children using high contrast stimuli.

Nonquadratic penalization improves near-infrared diffuse optical tomography

A new approach that can easily incorporate any generic penalty function into the diffuse optical tomographic image reconstruction is introduced to show the utility of nonquadratic penalty functions. The penalty functions that were used include quadratic (l(2)), absolute (l(1)), Cauchy, and Geman-McClure. The regularization parameter in each of these cases was obtained automatically by using the generalized cross-validation method. The reconstruction results were systematically compared with each other via utilization of quantitative metrics, such as relative error and Pearson correlation. The reconstruction results indicate that, while the quadratic penalty may be able to provide better separation between two closely spaced targets, its contrast recovery capability is limited, and the sparseness promoting penalties, such as l(1), Cauchy, and Geman-McClure have better utility in reconstructing high-contrast and complex-shaped targets, with the Geman-McClure penalty being the most optimal one. (C) 2013 Optical Society of America

Image deconvolution research: its scope and importance in live cell microscopy

Fluorescence microscopy has become an indispensable tool in cell biology research due its exceptional specificity and ability to visualize subcellular structures with high contrast. It has highest impact when applied in 4D mode, i.e. when applied to record 3D image information as a function of time, since it allows the study of dynamic cellular processes in their native environment. The main issue in 4D fluorescence microscopy is that the phototoxic effect of fluorescence excitation gets accumulated during 4D image acquisition to the extent that normal cell functions are altered. Hence to avoid the alteration of normal cell functioning, it is required to minimize the excitation dose used for individual 2D images constituting a 4D image. Consequently, the noise level becomes very high degrading the resolution. According to the current status of technology, there is a minimum required excitation dose to ensure a resolution that is adequate for biological investigations. This minimum is sufficient to damage light-sensitive cells such as yeast if 4D imaging is performed for an extended period of time, for example, imaging for a complete cell cycle. Nevertheless, our recently developed deconvolution method resolves this conflict forming an enabling technology for visualization of dynamical processes of light-sensitive cells for durations longer than ever without perturbing normal cell functioning. The main goal of this article is to emphasize that there are still possibilities for enabling newer kinds of experiment in cell biology research involving even longer 4D imaging, by only improving deconvolution methods without any new optical technologies.

Advances in Image Reconstruction Methods for Real-Time Magnetic Resonance Thermometry

Magnetic Resonance Imaging (MRI) has been widely used in cancer treatment planning, which takes the advantage of high-resolution and high-contrast provided by it. The raw data collected in the MRI can also be used to obtain the temperature maps and has been explored for performing MR thermometry. This review article describes the methods that are used in performing MR thermometry, with an emphasis on reconstruction methods that are useful to obtain these temperature maps in real-time for large region of interest. This article also proposes a prior-image constrained reconstruction method for temperature reconstruction in MR thermometry, and a systematic comparison using ex-vivo tissue experiments with state of the art reconstruction method is presented.

Limited-view light sheet fluorescence microscopy for three dimensional volume imaging

We propose and demonstrate a limited-view light sheet microscopy (LV-LSM) for three dimensional (3D) volume imaging. Realizing that longer and frequent image acquisition results in significant photo-bleaching, we have taken limited angular views (18 views) of the macroscopic specimen and integrated with maximum likelihood (ML) technique for reconstructing high quality 3D volume images. Existing variants of light-sheet microscopy require both rotation and translation with a total of approximately 10-fold more views to render a 3D volume image. Comparatively, LV-LSM technique reduces data acquisition time and consequently minimizes light-exposure by many-folds. Since ML is a post-processing technique and highly parallelizable, this does not cost precious imaging time. Results show noise-free and high contrast volume images when compared to the state-of-the-art selective plane illumination microscopy. (C) 2015 AIP Publishing LLC.

HOMOGENIZATION OF A HYPERBOLIC EQUATION WITH HIGHLY CONTRASTING DIFFUSIVITY COEFFICIENTS

We study a hyperbolic problem in the framework of periodic homogenization assuming a high contrast between the diffusivity coefficients of the two components M-epsilon and B-epsilon of the heterogeneous medium. There are three regimes depending on the ratio between the size of the period and the amplitude a, of the diffusivity in B-epsilon. For the critical regime alpha(epsilon) similar or equal to epsilon, the limit problem is a strongly coupled system involving both the macroscopic and the microscopic variables. We also include the results in the non critical case.

HOMOGENIZATION OF A HYPERBOLIC EQUATION WITH HIGHLY CONTRASTING DIFFUSIVITY COEFFICIENTS

We study a hyperbolic problem in the framework of periodic homogenization assuming a high contrast between the diffusivity coefficients of the two components M-epsilon and B-epsilon of the heterogeneous medium. There are three regimes depending on the ratio between the size of the period and the amplitude a, of the diffusivity in B-epsilon. For the critical regime alpha(epsilon) similar or equal to epsilon, the limit problem is a strongly coupled system involving both the macroscopic and the microscopic variables. We also include the results in the non critical case.

Ultrafast control of multiple filamentation by ultrafast laser pulses

Control of multiple filamentation by laser-induced microlens effect due to a nonlinear interaction of two overlapping laser beams inside a glass plate was demonstrated. Individual or multiple spots on the white light pattern which is a product of multiple filamentation through a mesh can be switched on and off with a very high contrast ratio on a femtosecond time scale. This phenomenon can find applications such as ultrafast optical switch and high-speed sampling. (C) 2005 American Institute of Physics.

利用块状介质进行飞秒强激光脉冲的腔外压缩

高强度飞秒激光脉冲的腔外压缩是获得高次谐波阿秒脉冲驱动源的必要手段。实验研究了超强超短飞秒激光脉冲在经过块状介质后的光谱展宽和色散补偿压缩现象。单脉冲能量0．26mJ，脉宽50fs的激光脉冲经透镜在空气中聚焦后再入射到块状材料上，出射脉冲光谱被展宽到接近40nm。由于在块状材料中的自聚焦效应，出射光束质量变好并保持较小的空间啁啾。利用熔融石英棱镜对补偿带有正色散的出射脉冲，最后得到〉0．1mJ，19fs的压缩脉冲。利用SPIDER装置测量了出射脉冲的脉宽和光谱相位。整个系统的能量效率大约为35％，压缩后的

基于孔径耦合带状线的激光脉冲整形系统

实现了一种新型的激光脉冲整形系统，该系统使用了一个由孔径耦合带状线（ACSL）电脉冲整形器驱动的电光调制器。一个电脉冲整形器由两条通过其公共接地板上的耦合孔径发生耦合作用的带状传输线所组成的四端口装置。更换具有不同耦合孔径的公共接地板，该电脉冲整形发生器可以具有150ps时间结构的任意整形电脉冲。将任意整形的电脉冲输入到电光调制器上，就可以得到任意整形的激光脉冲。利用该系统，激光脉冲整形系统能够产生具有150ps前后沿，1～3ns脉冲宽度可调、高对比度、光滑过渡以及任意整形的激光脉冲。

高功率激光装置前端系统激光脉冲的时间整形及修正技术

针对激光惯性约束聚变实验研究对高功率激光驱动器前端系统复杂时间形状种子激光脉冲的需求, 应用孔径耦合带状线集成波导整形系统设计了满足需要的前端整形激光脉冲。用一种新方法精确计算了孔径耦合带状线电脉冲整形器的耦合系数和孔径宽度的数值关系, 并针对高衬比度整形激光脉冲的需求, 提出了高衬比度双极型集成波导整形系统方案。由该系统可以得到100 ps脉冲前沿、1～3 ns脉冲宽度可调、高衬比度(大于100∶1)、光滑无纹波调制、可精确满足神光II八路及第九路装置需求的前端整形激光脉冲。