Optical sectioning microscope with a binary hologram based beam scanning.

We describe the development of a beam scanning microscope that can perform optical sectioning based on the principle of confocal microscopy. The scanning is performed by a laser beam diffracted from a dynamic binary hologram implemented using a liquid crystal spatial light modulator. Using the proposed scanning mechanism, unlike the conventional confocal microscopes, scanning over a two-dimensional area of the sample can be obtained without the use of a pair of galvo mirror scanners. The proposed microscope has a number of advantages, such as superior frame to frame repeatability, simpler optical arrangement, increased pixel dwell time relative to the time between two pixels, illumination of only the sample points without pulsing the laser, and absolute control over the amplitude and phase of the illumination beam on a pixel to pixel basis. The proposed microscope can be particularly useful for applications requiring very long exposure time or very large working distance objective lenses. In this paper we present experimental implementation of the setup using a nematic liquid crystal spatial light modulator and proof-of-concept experimental results.

Lateral resolution enhancement in confocal microscopy by vectorial aperture engineering.

This article reports the design and implementation of a lateral resolution-enhancement technique in confocal microscopy that can work, in principle, either in the reflection mode or in the fluorescence mode. Taking the difference between two images corresponding to two different vectorially (involving amplitude, phase, and polarization of light) engineered illumination pupils or apertures of a confocal microscope, high spatial frequency contents in the resultant image can be significantly enhanced. This can be realized by incorporating an extra vectorial beam-forming element into the illumination beam path of a conventional confocal microscope. The method of the proposed technique has been explained by giving it an analytical treatment supported by numerical simulation results. The technique has been implemented in a reflection mode confocal microscope and results obtained are presented.

Zonal wavefront sensing using an array of gratings.

We describe a zonal-wavefront-sensing technique using an array of plane diffraction gratings. A spatially coherent beam, whose wavefront is to be measured, is incident on the array of gratings. The direction of a diffracted beam of a certain diffraction order is a function of the orientation and periodicity of the corresponding grating. Thus, by choosing the orientation and periodicity of each grating appropriately and by having a lens immediately behind the grating array, it is possible to get an array of focal spots. The profile of the incident wavefront can be estimated from the displacements of these focal spots relative to those due to an unaberrated beam. The arrangement makes it possible to increase the separation between two adjacent focal spots corresponding to two nearby gratings without effecting the areas of the gratings. Consequently, a relatively large dynamic range in wavefront measurement can be achieved without compromising the accuracy. With the arrangement it is also possible to use a photodetector array whose outline is independent of the grating array outline. The proposed wavefront-sensing technique is implemented experimentally using a liquid-crystal spatial-light modulator in conjunction with a CCD camera, and the obtained results are presented.

Laser scanning confocal microscope with programmable amplitude, phase, and polarization of the illumination beam.

We describe the design and construction of a laser scanning confocal microscope with programmable beam forming optics. The amplitude, phase, and polarization of the laser beam used in the microscope can be controlled in real time with the help of a liquid crystal spatial light modulator, acting as a computer generated hologram, in conjunction with a polarizing beam splitter and two right angled prisms assembly. Two scan mirrors, comprising an on-axis fast moving scan mirror for line scanning and an off-axis slow moving scan mirror for frame scanning, configured in a way to minimize the movement of the scanned beam over the pupil plane of the microscope objective, form the XY scan unit. The confocal system, that incorporates the programmable beam forming unit and the scan unit, has been implemented to image in both reflected and fluorescence light from the specimen. Efficiency of the system to programmably generate custom defined vector beams has been demonstrated by generating a bottle structured focal volume, which in fact is the overlap of two cross polarized beams, that can simultaneously improve both the lateral and axial resolutions if used as the de-excitation beam in a stimulated emission depletion confocal microscope.

Minerals and functional groups present in the jackfruit seed: a spectroscopic investigation.

In this work, emission and Fourier transform infrared spectra of the jackfruit seed, in powdered form, are recorded. Analysis of the emission spectrum confirms the presence of two hitherto undetected elements, manganese and magnesium. The Fourier transform infrared spectrum reveals the presence of some specific functional groups, attributed to the different bands present in the spectrum.