In situ measurement of three-dimensional ion densities in focused femtosecond pulses

We image spatial distributions of Xeq+ ions in the focus of a laser beam of ultrashort, intense pulses in all three dimensions, with a resolution of approximately 3 microm and approximately 12 microm in the two transverse directions. This allows for studying ionization processes without spatially averaging ion yields. Our in situ ion imaging is also useful to analyze focal intensity profiles and to investigate the transverse modal purity of tightly focused beams of complex light. As an example, the intensity profile of a Hermite-Gaussian beam mode HG1,0 recorded with ions is found to be in good agreement with optical images.     

Transverse superresolution and focal shift with rotational tunable phase mask

This paper reports the imaging characteristics of an optical system can be modified by our designed polarization pupil mask. The novel rotational symmetric polarization pupil mask design based on combination of half-wave and quarter-wave plates is introduced for realizing the focal shift and extending focal depth of an optical system and the procedure for designing is presented. Numerical results show when an appropriate rotational symmetric polarization pupil mask is used as an apodizer in the optical imaging system, it not only can effectively achieve the continuously focal shift in a small range and extend focal depth of the optimized system, but also can evidently increase the transverse resolution of the optimized system at the genuine focal plane.     

Adaptive-optics ultrahigh-resolution optical coherence tomography

Merging of ultrahigh-resolution optical coherence tomography (UHR OCT) and adaptive optics (AO), resulting in high axial (3 microm) and improved transverse resolution (5-10 microm) is demonstrated for the first time to our knowledge in in vivo retinal imaging. A compact (300 mm x 300 mm) closed-loop AO system, based on a real-time Hartmann-Shack wave-front sensor operating at 30 Hz and a 37-actuator membrane deformable mirror, is interfaced to an UHR OCT system, based on a commercial OCT instrument, employing a compact Ti:sapphire laser with 130-nm bandwidth. Closed-loop correction of both ocular and system aberrations results in a residual uncorrected wave-front rms of 0.1 microm for a 3.68-mm pupil diameter. When this level of correction is achieved, OCT images are obtained under a static mirror configuration. By use of AO, an improvement of the transverse resolution of two to three times, compared with UHR OCT systems used so far, is obtained. A significant signal-to-noise ratio improvement of up to 9 dB in corrected compared with uncorrected OCT tomograms is also achieved.     

Femtosecond micromachining of symmetric waveguides at 1.5 microm by astigmatic beam focusing

We report on a new spatial beam-shaping approach for fabrication of waveguides with a circular transverse profile by femtosecond laser pulses, using an astigmatic beam and controlling both beam waist and focal position in the tangential and sagittal planes. We apply this technique to write single-mode active waveguides at 1.5microm in Er:Yb-doped glass substrates. The experimental results are well described by a simple nonlinear absorption model.     

A conical air-coupled capacitance transducer for surface imaging

This paper describes the construction and operation of an air-coupled capacitance transducer with a conical backplate. This was designed to produce a focal region over an extended distance along the transducer axis. Measurements were performed to examine both the frequency response of the transducer in pulse-echo mode, and the lateral resolution for imaging purposes. The radiated field was measured and compared to theory, and the extent of the focal region determined. Images of surface topography are presented, to illustrate the range of application of the transducer.     

Single-body lensed photonic crystal fibers as side-viewing probes for optical imaging systems

We report the fabrication and performance of a lensed photonic crystal fiber (PCF) designed as a compact but effective side-viewing optical imaging probe. The lensed-PCF probe was implemented in a single body without using any other fibers or additional optics. The beam expansion region and a focusing ball lens, necessary for a focuser, were simultaneously formed along a small piece of PCF by applying arc discharges. The side-viewing ability was provided by polishing the ball lens with a femtosecond laser to form a total internal reflection surface. The working distance and the transverse resolution of the fabricated single-body lensed-PCF were experimentally measured to be 570 and 6.8 microm, respectively. With the proposed lensed-PCF probe, optical coherence tomography images of an in vitro biological sample were successfully obtained.     

Dual-axis confocal microscope for high-resolution in vivo imaging

We describe a novel confocal microscope that uses separate low-numerical-aperture objectives with the illumination and collection axes crossed at angle theta from the midline. This architecture collects images in scattering media with high transverse and axial resolution, long working distance, large field of view, and reduced noise from scattered light. We measured transverse and axial (FWHM) resolution of 1.3 and 2.1 microm, respectively, in free space, and confirm subcellular resolution in excised esophageal mucosa. The optics may be scaled to millimeter dimensions and fiber coupled for collection of high-resolution images in vivo.     

Large area confocal microscopy

Imaging large tissue areas with microscopic resolution in vivo may offer an alternative to random excisional biopsy. We present an approach for performing confocal imaging of large tissue surface areas using spectrally encoded confocal microscopy (SECM). We demonstrate a single-optical-fiber SECM apparatus, designed for imaging luminal organs, that is capable of imaging with a transverse resolution of 2.1 microm over a subsurface area of 16 cm2 in less than 1 min. Due to the unique probe configuration and scanning geometry, the speed and resolution of this new imaging technology are sufficient for comprehensively imaging large tissues areas at a microscopic scale in times that are appropriate for clinical use.     

Real-time two-dimensional imaging in scattering media by use of a femtosecond Cr(4+):forsterite laser

An original femtosecond Cr(4+):forsterite laser source associated with a nonlinear optical correlator was used for imaging through scattering media with 1220-nm light. The system, which operates as an ultrafast optical gate by sum-frequency generation in a nonlinear crystal, was able to detect the light reflected from a resolution chart hidden in a turbid medium, at an attenuation of as much as 15 mean free paths. When the object was illuminated with a collimated beam, real-time two-dimensional images were obtained, with a maximum transverse resolution of ~20 microm.     

Extended depth of focus in tomographic phase microscopy using a propagation algorithm

Tomographic phase microscopy is a laser interferometry technique in which a 3D refractive index map of a biological sample is constructed from quantitative phase images collected at a set of illumination angles. Although the resulting tomographic images provide valuable information, their resolution declines at axial distances beyond about 1 microm from the focal plane. We describe an improved 3D reconstruction algorithm in which the field at the focal plane is numerically propagated to depths throughout the sample. Diffraction is thus incorporated, extending the depth of focus to more than 10 mum. Tomograms with improved focal depth are demonstrated for single HT29 cells.     

Toward a spherical spot distribution with 4pi focusing of radially polarized light

The properties of the focal spot for 4pi focusing with radially polarized light are presented for various apodization factors. With a focusing system satisfying the Herschel condition, sharp focal spots with almost-perfect spherical symmetry (leading to equal axial and transverse resolution) and extremely low sidelobes are achieved.     

Depth-resolved holographic imaging through scattering media by use of a photorefractive polymer composite device in the near infrared

We report full-field depth-resolved holographic imaging through scattering media by use of a photorefractive polymer device in the near infrared. Real-time images through seven scattering mean free paths are acquired in a few seconds with a broadband superluminescent diode at 794 nm. A depth resolution of 15 microm and a transverse resolution of better than 42 microm are achieved. The photorefractive devices used are based on a poly(N-vinylcarbazole):2,4,7-trinitro-9-fluorenone dimalenitrile charge-transport network doped with the electro-optic chromophore 1-(2'-ethylhexyloxy)-2,5-dimethyl-4-(4"-nitrophenylazo)benzene.     

In vivo fluorescent imaging of the mouse retina using adaptive optics

In vivo imaging of the mouse retina using visible and near infrared wavelengths does not achieve diffraction-limited resolution due to wavefront aberrations induced by the eye. Considering the pupil size and axial dimension of the eye, it is expected that unaberrated imaging of the retina would have a transverse resolution of 2 microm. Higher-order aberrations in retinal imaging of human can be compensated for by using adaptive optics. We demonstrate an adaptive optics system for in vivo imaging of fluorescent structures in the retina of a mouse, using a microelectromechanical system membrane mirror and a Shack-Hartmann wavefront sensor that detects fluorescent wavefront.     

Analysis of the healthy rabbit lens surface using MAC Mode atomic force microscopy

In this investigation healthy rabbit crystalline lenses were characterized by atomic force microscopy (AFM). The lenses were cut in slices with thickness with 1mm and thus, put after cortex distinct regions of nucleus and cortex for AFM examination. AFM analysis were carried out using a PicoSPM I operating in Mac Mode. We obtained topographic images of rabbit lenses and a quantitative analysis of the width and height of fibers for nucleus and cortex regions. The longitudinal section analysis of fibers in the nucleus region indicated structures with an average width of 200nm and average height of 200nm. The intershells distance was determined as 4microm. Fiber cell cross-section dimensions, longitudinal and transverse widths, could be estimated in these regions from the AFM images. Structures with average widths as small as 1.0microm are observed in the nucleus; the intershell distance is 4.0microm. In cortical regions, hexagonal structures with average longitudinal and transverse widths of 5.0mum and 3.0mum, respectively, were identified. Three-dimensional images of tissue sections with resolution on a nanometer scale were obtained. The potential of AFM analysis for characterizing healthy and pathologic lens tissues is discussed.     

Active focus locking in an optically sectioning microscope utilizing a deformable membrane mirror

A significant challenge for in vivo imaging is to remove movement artifacts. These movements (typically due to either respiration and cardiac-related movement or surface chemical response) are normally limited to the axial direction, and hence features move in and out of the focal plane. This presents a real problem for high-resolution optically sectioned imaging techniques such as confocal and multiphoton microscopy. To overcome this we have developed an actively locked focus-tracking system based around a deformable membrane mirror. This has a significant advantage over more conventional focus-tracking techniques where the microscope objective is dithered, since the active element is not in direct, or indirect, contact with the sample. To examine the operational limits and to demonstrate possible applications for this form of focus locking, sample oscillation and movement are simulated for two different biological applications. We were able to track focus over a 400 microm range (limited by the range of the piezomounted objective) with a rms precision on the focal depth of 0.31 microm +/- 0.05 microm.     

Slanted hole array beam profiler (SHArP)-a high-resolution portable beam profiler based on a linear aperture array

We demonstrate a novel high-resolution portable beam profiler based on a slanted linear array of small apertures, termed a slanted hole array beam profiler (SHArP). The apertures are directly fabricated on a metal-coated CMOS imaging sensor. With a single linear scan, the aperture array can establish a virtual grid of sampling points for beam profiling. With our prototype, we demonstrate beam profiling of Gaussian beams over an area of 66.5 microm x 66.5 microm with a resolution of 0.8 microm (compare with the CMOS pixel size of 10 microm). The resolution can be improved into the range of submicrometers by fabricating smaller apertures. The good correspondence between the measured and calculated beam profiles proves the fidelity of our new beam profiling scheme.     

Microtubule dynamics depart from the wormlike chain model

Thermal shape fluctuations of grafted microtubules were studied using high resolution particle tracking of attached fluorescent beads. First mode relaxation times were extracted from the mean square displacement in the transverse coordinate. For microtubules shorter than approximately 10 microm, the relaxation times were found to follow an L2 dependence instead of L4 as expected from the standard wormlike chain model. This length dependence is shown to result from a complex length dependence of the bending stiffness which can be understood as a result of the molecular architecture of microtubules. For microtubules shorter than approximately 5 microm, high drag coefficients indicate contributions from internal friction to the fluctuation dynamics.     

Tracking optical coherence tomography

An experimental tracking optical coherence tomography (OCT) system has been clinically tested. The prototype instrument uses a secondary sensing beam and steering mirrors to compensate for eye motion with a closed-loop bandwidth of 1 kHz and tracking accuracy, to within less than the OCT beam diameter. The retinal tracker improved image registration accuracy to <1 transverse pixel (<60 microm). Composite OCT images averaged over multiple scans and visits show a sharp fine structure limited only by transverse pixel size. As the resolution of clinical OCT systems improves, the capability to reproducibly map complex structures in the living eye at high resolution will lead to improved understanding of disease processes and improved sensitivity and specificity of diagnostic procedures.     

Enhanced background rejection in thick tissue using focal modulation microscopy with quadrant apertures

Image formation in focal modulation microscopy using quadrant apertures (QFMM) is presented. The spatial resolution is discussed: compared with confocal microscopy, QFMM can simultaneously enhance the axial and transverse resolution. In contrast to focal modulation microscopy using D-shaped apertures (DFMM), QFMM can maintain x-y symmetry of spatial resolution, thus causing less confusion in imaging interpretation. The capability of background rejection is investigated, showing QFMM has the potential to increase imaging penetration depth. The signal level is also analyzed, indicating that the preferred detector pinhole radius is greater than for DFMM, resulting in a higher signal level.     

三环位相型光瞳滤波器的横向超分辨与轴向焦深扩展

用矢量衍射方法分析了线偏振光入射到带有三环位相型光瞳滤波器的高数值孔径物镜时,焦点的轴向和横向光强分布.数值模拟结果表明,高数值孔径物镜聚焦时需同时考虑光强的轴向和横向分布.通过加入三环位相光瞳滤波器,在实现横向超分辨的同时实现了光学系统轴向焦深扩展和轴向光强分布平坦化,并且位相调制深度变化时,会出现轴向焦移现象.对三环位相光瞳滤波器结构进行了优化,得出了优化结果. 关键词： 光学超分辨 扩展焦深 位相型光瞳滤波器 矢量衍射方法