John M. Girkin

Practical implementation of adaptive optics in multiphoton microscopy

A dedicated two-photon microscope incorporating adaptive-optic correction of specimen-induced aberrations is presented. Wavefront alteration of the scanning laser beam was achieved by use of a micromachined deformable mirror. Post scan head implementation produces a compact module compatible with the Bio-Rad MRC-600 scan head. Automatic aberration correction using feedback from the multiphoton fluorescence intensity allowed the adaptive optic to extend the imaging depth attainable in both artificial and biological refractive-index mismatched samples. With a 1.3-NA, x40, Nikon oil immersion objective, the imaging depth in water was extended from approximately 3.4 to 46.2 microm with a resolution defined by a FWHM axial point-spread function of 1.25 microm.

Optical ferris wheel for ultracold atoms

We propose a versatile optical ring lattice suitable for trapping cold and quantum degenerate atomic samples. We demonstrate the realisation of intensity patterns from pairs of Laguerre-Gauss (exp(i??) modes with different ? indices. These patterns can be rotated by introducing a frequency shift between the modes. We can generate bright ring lattices for trapping atoms in red-detuned light, and dark ring lattices suitable for trapping atoms with minimal heating in the optical vortices of blue-detuned light. The lattice sites can be joined to form a uniform ring trap, making it ideal for studying persistent currents and the Mott insulator transition in a ring geometry.

GaN micro-light-emitting diode arrays with monolithically integrated sapphire microlenses

GaN micro-light-emitting diodes (micro-LEDs) with monolithically integrated microlenses have been demonstrated. Microlenses, with a focal length of 44 μm and a root mean square roughness of ∼1 nm, have been fabricated on the polished back surface of a sapphire substrate of an array of micro-LEDs by resist thermal reflow and plasma etching. The optical properties of the microlenses have been demonstrated to alter the emission pattern of the LED emitters. The cone of light emitted from this hybrid device is significantly less divergent than a conventional broad-area device. This combination of micro-LED and microlens technologies offers the potential for further improvement in the overall efficiency of GaN-based light emitters.

Adaptive optics for enhanced signal in CARS microscopy.

We report the use of adaptive optics with coherent anti-Stokes Raman scattering (CARS) microscopy for label-free deep tissue imaging based on molecular vibrational spectroscopy. The setup employs a deformable membrane mirror and a random search optimization algorithm to improve signal intensity and image quality at large sample depths. We demonstrate the ability to correct for both system and sample-induced aberrations in test samples as well as in muscle tissue in order to enhance the CARS signal. The combined system and sample-induced aberration correction increased the signal by an average factor of approximately 3x for the test samples at a depth of 700 microm and approximately 6x for muscle tissue at a depth of 260 microm. The enhanced signal and higher penetration depth offered by adaptive optics will augment CARS microscopy as an in vivo and in situ biomedical imaging modality.

Surface-enhanced Raman scattering spectroscopy as a sensitive and selective technique for the detection of folic acid in water and human serum.

Surface-enhanced Raman scattering (SERS) is shown to give linear and sensitive concentration-dependent detection of folic acid using silver nanoparticles created via ethylene-diaminetetraacetic acid (EDTA) reduction. Optical detection by SERS overcomes the primary limitation of photodissociation encountered during the application of other shorter wavelength ultraviolet (UV)/near-UV techniques such as fluorescence based microscopy. The SERS approach in water-based samples was demonstrated and optimized using several longer wavelengths of excitation (514.5, 632.8, and 785 nm). Excitation in the green (514.5 nm) was found to achieve the best balance between photodissociation and SERS efficiency. Linear concentration dependence was observed in the range of 0.018 to 1 μM. The importance of folic acid in a clinical setting and the potential applications of this technique in a biological environment are highlighted. We demonstrate the potential to transfer this technique to real biological samples by the detection of folic acid in human serum samples by SERS.

Creating permanent 3D arrangements of isolated cells using holographic optical tweezers

We report the creation of permanent 3D configurations of cells, at predefined positions, within a gelatin matrix. The technique used holographic optical tweezers to manipulate individual E. coli within a solution comprising monomer precursors. The matrix was then set and after the laser beam was removed, we were able to demonstrate that the structures remained intact for many days. We were also able to demonstrate that, in the presence of appropriate nutrients, the E. coli survived within the gelatin matrix for several days. The technique could have a number of potential future applications, including the arrangement of a variety of different cell types in complex architectures, as motifs for promoting tissue differentiation and growth within the field of cell engineering.

Adaptive optics for deeper imaging of biological samples.

Optical microscopy has been a cornerstone of life science investigations since its first practical application around 400 years ago with the goal being subcellular resolution, three-dimensional images, at depth, in living samples. Nonlinear microscopy brought this dream a step closer, but as one images more deeply the material through which you image can greatly distort the view. By using optical devices, originally developed for astronomy, whose optical properties can be changed in real time, active compensation for sample-induced aberrations is possible. Submicron resolution images are now routinely recorded from depths over 1mm into tissue. Such active optical elements can also be used to keep conventional microscopes, both confocal and widefield, in optimal alignment.

Reflection/transmission confocal microscopy characterization of single-crystal diamond microlens arrays

Using the method of photoresist reflow and inductively coupled plasma dry etching, we have fabricated microlens arrays in type-IIa natural single-crystal diamond, with diameters down to 10 μm. The surface profile of the microlenses was characterized by atomic force microscopy and was found to match well with a spherical shape, with a surface roughness of better than 1.2 nm. To characterize the optical properties of these diamond microlens arrays, a laser scanning reflection/transmission confocal microscopy technique has been developed. This technique enabled the surface profile of the microlenses to be measured simultaneously with optical parameters including focal length and spot size, opening up an application area for confocal microscopy.

Generation of achromatic Bessel beams using a compensated spatial light modulator.

We report the creation of white-light, achromatic Bessel beams using a spatial light modulator and a prism to compensate for the dispersion. Unlike the Bessel beam created by a refractive axicon, this achromatic beam has a radial wavevector and hence an intensity cross-section which is independent of wavelength. The technique also lends itself to the generation of higher order Bessel beams with an on-axis optical vortex and associated orbital angular momentum.

Evaluation of enamel dental restoration interface by optical coherence tomography

Evaluation of molar dental restorations on enamel is performed using optical coherence tomography (OCT) with 10 microm resolution. Images of approximately 50 microm failure gaps in the restorations are demonstrated and the OCT images are compared with x-ray and optical microscopy pictures. The results demonstrate the potential of the technique for clinical evaluation of dental restorations.