Nanophotonics has emerged as a new and important field of study, not only in research, but also in undergraduate optics and photonics education and training. Beyond the study of classical and quantum optics, it is important for students to learn about how the flow of light can be manipulated on a nanoscale level, and used in applications such as telecommunications, imaging, and medicine. This paper reports on our work to integrate basic nanophotonic concepts and topics into existing optics and optical electronics courses, as well as independent study projects, at the undergraduate level. Through classroom lectures, topical readings, computer modeling exercises, and laboratory experiments, students are introduced to nanophotonic concepts subsequent to a study of physical and geometrical optics. A compare and contrast methodology is employed to help students identify similarities and differences that exist in the optical behavior of bulk and nanostructured media. Training is further developed through engineering design and simulation exercises that use advanced, vector-diffraction-based, modeling software for simulating the performance of various materials and structures. To date, the addition of a nanophotonics component to the optics curriculum has proven successful, been enthusiastically received by students, and should serve as a basis for further course development efforts that emphasize the combined capabilities of nanotechnology and photonics.
Subwavelength structures (SWS), a form of diffractive optic, are well-known for their ability to function as polarizers and anti-reflection coatings. They can also be used to create narrowband optical filters whose surface reflectance spectra exhibit resonant peaks that are highly wavelength dependent, especially as the surface index is modified by the deposition or adsorption of biomaterials such as molecules or cells. In this study, we report on the design and fabrication of SWS structures in silicon that are suitable for use as biosensors in sensitive molecular detection. The structures combine a two-dimensional dielectric grating and Si/SiO2 optical waveguide to create a surface that can function as a narrowband optical filter. The SWS structures were fabricated using a combination of three-beam interference lithography and reactive-ion etching in a CBrF3 plasma. This produced a two-dimensional periodic nanostructure grating array, having a period of ~450 nm and air pores of ~265 nm, within a 300 nm thick silicon layer that serves as the core waveguide region of the filter. The ability to achieve sensitive molecular detection (< nm) is expected by virtue of working with high-index silicon-based structures, but may be practically limited by the need to detect the reflectance at near-infrared, rather than visible, wavelengths.
The results of experimental studies that investigate the reactive ion etching characteristics of Si nanostructures using CBrF3 plasmas is reported. Etch rates and anisotropies were studied as functions of RF and ICP power (0 - 200 W), gas pressure (10 - 190 mT), gas flow, and O2 gas mixtures in a commercial ICP etching system. Both isotropic and anisotropic etching regimes were identified, and used to create < 600 nm feature periodic motheye and photonic crystal nanostructures that possess pyramidal and near- anisotropic profiles, respectively. The ability of CBrF3 to effectively control sidewall profile and etch depth when used with different resist mask profiles makes it an attractive etchant for the fabrication of periodic optical nanostructures that have stringent sidewall and nano-tolerance requirements.
Photonic crystals are emerging as an important class of engineered nanophotonic devices that possess unique optical properties and which can also provide textured surfaces for the study and control of cellular and molecular interactions. From among the many types of photonic crystal structures, two-dimensional (2D) and planar (slab) photonic crystals are the most attractive because of their ability to support guided-wave and active optical devices in semiconductor and polymer materials, serve as templates for device replication, and interface with colloidal and nanoparticle systems. This paper reports on the results of modeling and design efforts that show how 2d and slab silicon photonic crystals, based on their in-plane optical waveguiding and out-of-plane radiation properties, might be used to probe surface-bound cells and molecules or perform localized spectroscopy. The results of a parametric analysis show that photonic crystals comprised of high-index contrast materials (e.g. Si, air) are sensitive to geometric and material factors, potentially making them an effective medium to study molecular and cellular interactions critical to a number of biotechnological applications
T-cell contact with antigen-presenting B cells initiates an activation cascade which includes an increase in T-cell intracellular calcium and leads to T-cell proliferation and differentiation. We studied cell-cell contact requirements for T-cell activation using an optical trap to control the orientation of T-cell/B-cell pairs and fluorescence microscopy to measure subsequent T-cell(Ca2+)i response. B cells or beads coated with antibodies to the T- cell receptor are trapped with a titanium-sapphire laser and placed at different locations along the T-cell, which has a polarized appearance defined by the shape and direction of crawling. T-cell (Ca2+)i is detected as an emission shift from the combination of fura-red and oregon- green, two cytoplasmic (Ca2+)i indicators. T- cells which are presented antigen at the leading edge have a higher probability of responding and a shorter latency of response than those contacting B-cells or beads with their trailing end.
Optical laser trapping microscopy has emerged as a powerful tool not only for the optical manipulation of cells and macromolecules, but also for the study of cellular physiological responses via force transduction and fluorescence imaging. We describe here the most recent results from our laboratory in the use and application of laser trapping microscopy to a variety of studies at the cellular and molecular levels. Fluorescence spectroscopy and imaging have been successfully combined with optical micromanipulation. A single near-infrared laser beam is used for two-photon fluorescence excitation and micromanipulation of trapped biological specimens. Cell viability is observed and monitored with a Nd:YAG laser ((lambda) equals 1064 nm) and an Al:GaAs diode laser ((lambda) equals 809 nm). Traps and conventional fluorescence imaging are also used simultaneously to examine T-cell activation dynamics.
The behavior of real scattering surfaces is often specified by measuring the bidirectional reflectance factor (BRF), defined as the ratio of the flux scattered into a given direction by a surface under given conditions of illumination to the flux scattered in the same direction by a Lambertian scatterer under identical conditions.THe utility of this factor is that measurements on surfaces can be related to known standards, which have a BRF greater than 99 percent for a broad range of wavelengths. In addition to the incidence angle and spectral features of the incident flux, the reflectance properties of a surface are affected by the intrinsic composition and roughness properties of the surface. Therefore, the spectral reflectance of different targets will generally yield spectral reflectance curves of different shapes, forming the basis for identification of materials. For example, optical principles developed for the determination of reflectance properties of marine particles facilitate the determination of the BRF of oceanic samples. We have recently developed and implemented a system for determining the BRF composed of a Zeiss photomicroscope equipped with a reflective system. In this system, excitation is provided over a large field of view while reflection collection is acquired over a slightly smaller solid angle. Multi-wavelength measurements allow the determination of the effect of the excitation wavelength on both the reflectance and fluorescence properties of the sample, whereas monochromatic measurements exclude fluorescence effects. This new technique provides the advantages of determination of the BRF for different types of individual and bulk particulates transferred onto an optical embedding medium or collected on an Anopore filter. Abundance and other optical properties of dominant particle types can also be determined by individual particle analysis on the same sample.
We report the implementation of an optical laser trap that incorporates phase conjugate microscopy as one of its key elements. The laser trap consists of a primary trapping beam, and a counterpropagating self-pumped phase conjugate beam, the latter being derived from the primary signalbeam when it is transmitted through the sample and phase conjugated in a photorefractive crystal. Using a fundamental TEM00 gaussian beam from an Ar-ion laser (514 nm), and a barium titanate (BaTiO3) crystal in the CAT (total internal reflection) geometry, an optical laser trap was created using approximately 25 mW of laser power. Strong transverse optical confinement is reported for approximately 2.5 micrometers diameter polystyrene microspheres, using both high and low numerical aperture (e.g. 60 X, 0.85 N.A. and 10 X, 0.25 N.A.) objective lenses. By virtue of the phase conjugation process, the present geometry achieves self-alignment of the two counter-propagating beams, as well as photorefractive gain in the ratio of nearly 4:1. The system has the potential for implementing several novel image processing functions unique to the nonlinear phase conjugation process, on living cells confined to an optical laser trap, including aberration correction, contrast reversal, and novelty filtering, respectively.
We report the observation of two-photon fluorescence excitation and cell confinement, simultaneously, in a continuous-wave (cw) single-beam gradient force optical trap, and demonstrate its use as an in-situ probe to study the physiological state of an optically confined cell sample. At the wavelength of 1064 nm, a single focused gaussian laser beam is used to simultaneously confine, and excite visible fluorescence from, a human sperm cell that has been tagged with propidium iodide, a exogenous fluorescent dye that functions as a viability assay of cellular physiological state. The intensity at the dye peak emission wavelength of 620 nm exhibits a near-square-law dependence on incident trapping beam photon laser power, a behavior consistent with a two-photon absorption process. In addition, for a sperm cell held stationary in the optical tweezers for a period of several minutes at a constant trapping power, red fluorescence emission was observed to increase the time, indicating that the cell has gradually transitioned between a live and dead state. Two-photon excited fluorescence was also observed in Chinese hamster ovary cells that were confined by cw laser tweezers and stained with either propidium iodide or Snarf, a pH-sensitive dye probe. These results suggest that, for samples suitably tagged with fluorescent probes and vital stains, optical tweezers can be used to generate their own in-situ diagnostic optical probes of cellular viability or induced photodamage, via two-photon processes.
We report on the development of a flow system integrated with a microscope that facilitates the simultaneous optical trapping and fluorescence excitation/detection of micron-sized samples that have been tagged with fluorescent probes. This system, when used in conjunction with nucleotide fluorescence labeling and DNA fragment cleavage procedures, offers the potential for the rapid sequencing of DNA fragments attached to microsphere 'handles.' Using a Nd:YAG laser (1064 nm) as the trapping beam, latex microspheres were stably trapped in a flow stream, where flow velocities in the range of 1 - 10 mm/s were achieved. Such flow velocities are commensurate with those required to stretch, and measure fluorescence from, DNA strands in high speed sequencing applications. High spatial resolution (approximately 1 micrometer) and high signal-to-noise ratios (greater than 103) were achieved using a high N.A. objective lens for trapping and fluorescence detection within a confocal microscope geometry. In this system, samples could be displaced with the scanning laser trap by up to plus or minus 25 micrometers off the trapping beam axis in the sample plane while maintaining, at the same time, a large fluorescence detection efficiency. At a trapping depth of 20 micrometer below the chamber surface, a laser power of 100 mW was sufficient to hold a 2 micrometer diameter microsphere in a flow stream having a velocity of 1 mm/s while its fluorescence was measured. The results of a systematic study which investigates the effects of trapping efficiency, trapping depth, flow velocity, and tweezers holding time in a 500 micrometer by 500 micrometer flow microchamber system are reported. The application of this technique to the confinement and detection of fluorescence-labeled DNA nucleotides is also described.
We describe fluorescence spectroscopy and imaging studies of optically trapped single Chinese hamster ovary (CHO) and motile human sperm cells. The NIR trapping beam was provided by a tunable, multimode continuous wave Ti:Sapphire laser. The beam was introduced into an inverted confocal laser scanning microscope. Fluorescence of cells in the single- beam gradient force optical trap was excited with a 488 nm microbeam (laser scanning microscopy) or with 365 nm radiation from a high- pressure mercury lamp. Modifications to NADH-attributed autofluorescence and Rhodamine- and Propidium Iodide-attributed xenofluorescence indicate a significant cell-damaging effect of 760 nm trapping beams. 760 nm effects produce a biological response comparable to UVA-induced oxidative stress and appear to be a consequence to two-photon absorption.
Microthermometric measurements on optically-trapped Chinese Hamster Ovary (CHO) cells and sperms cells re reported, using a noninvasive microfluorometric detection technique. Within an optical tweezer system that has been outfitted with a spectral fluorescence excitation and detection capability, the changes in temperature induced by the process of sample confinement by a focused laser beam has been quantified over micron-sized spatial regions of both motile and immotile cells. Our measurement technique is based on the use of environmentally sensitive fluorophores that can be incorporated into the cell membrane and used to sense local changes in temperature when the cell membrane is perturbed optically or via other environmental stress factors. Using a cw 1.064 micrometers Nd:YAG laser for trapping CHO and human sperm cells, a temperature increase of approximately equals 1°C per 100 mW laser power was observed. At this infrared wavelength, cellular heating as result of laser confinement appears to be mainly due to radiation absorption by water.
Optical trapping is a novel technique that utilizes radiation pressure for the non-contact manipulation and control of micron sized particles including living cells and micro-organisms. Typically, optical trapping is performed using a high power microscope and a complicated optical setup to form a single beam optical gradient trap or `optical tweezers.' Recent work has demonstrated three dimensional optical trapping using the counterpropagating beams from two optical fiber cleaves. This results in a simple and low cost implementation of an optical trap. Our paper discuses a refinement of this technique using pigtailed 1.3 micrometers semiconductor lasers and tapered lensed optical fibers with hemispherically machined microlens ends. Our optical trap consists of two tapered fiber lenses separated by distances of between 100 and 400 microns, with optical power ranges between 2 and 40 mW. Adjusting the relative powers of the optical fibers allowed us to trap and position 3, 5 and 10 micron beads over axial distances of several hundred microns. Our refinements improve trap accessibility while simultaneously increasing the trap stability. We have also used a ray optics model to simulate the performance of the optical fiber trap and predict the forces generated throughout the trapping volume. Axial and transverse trapping efficiencies up to 0.1 are predicted. The model can be used to predict trap strength and stability for various combinations of fiber spacings and particle sizes. Experimental observations of trapping and manipulation of 3 micrometers , 5 micrometers , and 10 micrometers beads are also presented and compared to the model.
Photoinduced modifications of NAD(P)H attributed autofluorescence of CHO cells in a single- beam gradient force optical trap (optical tweezers) were studied. Fluorescence spectra of single cells in the optical trap were measured using a modified microscope with an IR microbeam at 1064 and 760 nm for trapping, UVA radiation at 365 nm for fluorescence excitation, and an optical multichannel analyzer for spectral recording. No strong effect of the 1064 nm trapping beam on fluorescence intensity and spectral characteristics was found, even for power densities up to 70 MW/cm2. In contrast, 760 nm microirradiation resulted in a significant fluorescence increase, probably indicating cell damage due to absorption by heme- containing molecules. UVA exposure (1 W/cm2) of the trapped cells generated within seconds an initial fluorescence decrease, followed by a significant increase up to 5X of the value prior to irradiation. The UVA-induced modifications reflect NAD(P)H auto-oxidation and irreversible cell damage due to oxidative stress.
Optical tweezers is a term used to describe the optical force generation and confinement process by a highly focused laser beam. The forces exerted by the tweezer are sufficient to confine and move cells and particles without physical contact. When integrated with fluorescence or scattering detection, the laser tweezer can become a powerful instrument for the rapid characterization of the optical properties of isolated organic marine particulates and phytoplanktonic cells, from which bulk properties may be inferred. This technique offers the advantage of studying planktonic cells and organisms in their natural environment by confinement without immobilization, thereby preserving the spectral absorption and fluorescence properties of the samples under study. Herein, we report, for the first time, on the measurement of the spectral fluorescence and scattering of cyanobacteria and diatoms which have been confined by an optical tweezer. Preliminary data shows the characteristic emission peak from the chlorophyll (alpha) pigment (685 nm) for both samples, as well as spectral features that may be related to other photosynthetic pigments.
Low frequency noise measurements of n-p-n polyemitter bipolar transistors showing an increasing base current 1/f noise amplitude spanning seven orders of magnitude and increasing exponential dependence on biasing current between Ib and Ib2 with increasing emitter interfacial oxide continuity and decreasing emitter junction depth are described. Coupled with corresponding variations in linearized and large signal device characteristics, these trends lead to base voltage noise intensities that can both increase and decrease with increasing biasing current within the moderate 0.6 V < Vbe < 0.8 V biasing range. This behavior has not been previously attributed to the combination of fundamental base current noise and its noise voltage transfer function explicitly, but has importance to the assessment and accurate modeling of polyemitter transistor noise performance in shallow emitter transistors. The large range and high resolution of these features also imply that low frequency noise may be useful in characterizing the structure of the broken emitter interfacial oxide layer and its electrical properties.
We report the feasibility study of a versatile optical configuration consisting of a phase conjugate Michelson interferometer in conjunction with microscopic imaging optics for image processing and trapping of microscopic objects. Our test samples include phase gratings, amplitude gratings (i.e., Ronchi rulings), polystyrene microspheres, and biological samples such as liposomes and yeast cells. We have experimentally demonstrated (1) the novelty filtering feature which enhances the image of moving phase objects by suppressing the stationary background, (2) contrast reversal which is useful for the imaging of light absorbing (or scattering) particles, (3) the aberration correction capability of the system to enhance the image quality of microscopic objects embedded in or otherwise distorted by aberrators, and (4) optical trapping of polystyrene microspheres. The potential of using this technique for the manipulation and diagnosis of biological cells and tissues is discussed.
We describe the use of the optical trapping technique to the study of micron-sized particles and cell samples, from which scattering, particle size, and optical parametric data can be derived. Its application to the study of phytoplanktonic cells and other biogenic particles is also presented. In comparison to conventional flow cytometric or volume scattering measurement systems, an optical trap utilizes the radiation forces, derived from a highly focused laser beam, to confine the particle under study to an optical potential well. The optical trap, therefore, functions simultaneously as both a non-contact micromanipulator and microforce transducer. In addition, forward angle light scattering measurements can be made while the cell sample is held by the focused laser beam. Forward light scattering measurements and calculations for optically trapped spherical test particles and mammalian cells, under low power (< 10 mW), are presented for the cases when the laser beam spot size (omega) o is approximately r, (lambda) /2n < (omega) o < r, and (omega) o approximately (lambda) /2n, respectively, where r is the cell radius and n is the refractive index of the surrounding water medium. Scattering data over the range from approximately 0 degree(s) to 45 degree(s) is shown to be a sensitive function of beam radius, particle size, and relative refractive index. The optical trapping technique should prove to be a powerful tool in the study of the optical properties of marine cells and organisms, and their dependence on external optical stimuli.
A new inverted laser trapping microscope, combined with a linear diode array, has been developed to measure light scattering from a single cell over a large (up to approximately equals 45 degree(s)) angular range in real time. Diffraction profiles from 5 and 10 micrometers diameter polystyrene dielectric microspheres are in qualitative agreement with Lorenz-Mie calculations in terms of the separation and number of side-lobe features. Diffraction profiles from red blood cells (RBC), Chinese Hamster Ovary (CHO) cells, and liposomes exhibit features that can ultimately be correlated to their size and shape. Results obtained with this instrument can be used to determine the optical properties of the trapped cell, the location of the cell in the optical trap with respect to the laser focal point, and the forces acting on the cell.
The properties and development of low-loss (<1 dB/cm) gradedindex(GRIN) polymer microstructure waveguides (PMSWs) on various substrates, including semiconductors, conductors, insulators, and Ceramics,
are described. PMSW films can now be produced, uniformly, over substrate areas exceeding 400 cm2. The polymer materials exhibit wide transmission bandwidths (0.3 to 2.7 pm) and thermal stability characteristics
over a large dynamic temperature range ( -100°C to +180°C). Local sensitization techniques have been applied to PMSW films to produce planar, single and multiplexed, holographic gratings. Wavelength division
demultiplexing devices and single-wavelength 1-to-N fanout optical interconnects have also been realized. The polymer technology reported is well suited for optical interconnection, signal processing, communication,and sensing applications.
A five-channel wavelength-division demultiplexer (WDDM) is demonstrated that is fabricated in polymer microstructure waveguides and operates over a 100-nm bandwidth centered at 750 nm in the near-IR. The device has a maximum diffraction efficiency of about 50 percent at 730 nm, a spectral bandwidth of about 15 nm, and effectively utilizes the large optical transparency of the photolime gelatin polymer material at laser diode wavelengths.
Planar polymer lightguides ofpoly-(methyl methacrylate) (PMMA) are doped with a UV
sensitive material and an azo dye, sensitive for green or blue light (514, 488 nm) simultaneously.
The UV-sensitive material is used for recording a permanent waveguide pattern
while the azo dye is meant for a reversible light induced refractive index change. Although
the azo dye is partly destroyed by the UV light and the photochemical reaction during waveguide
formation, a light induced phase modulation in the guide is still possible in such
bifunctional materials.
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