Virtual optics and sensing of the retrieved complex field in the back focal plane using a constrained defocus algorithm.

The reflected back focal plane from a microscope objective is known to provide excellent information of material properties and can be used to analyze the generation of surface plasmons and surface waves in a localized region. Most analysis has concentrated on direct measurement of the reflected intensity in the back focal plane. By accessing the phase information, we show that examination in the back focal plane becomes considerably more powerful allowing the reconstructed field to be filtered, propagated and analyzed in different domains. Moreover, the phase often gives a superior measurement that is far easier to use in the assessment of the sample, an example of such cases is examined in the present paper. We discuss how the modified defocus phase retrieval algorithm has the potential for real time measurements with parallel image acquisition since only three images are needed for reliable retrieval of arbitrary distributions.

Spatially resolved random-access pump-probe microscopy based on binary holography.

In this Letter, we present a spatially resolved pump-probe microscope based on a digital micromirror device (DMD). The microscope system enables the measurements of ultrafast transient processes at arbitrarily selected regions in a 3-D specimen. To achieve random-access scanning, the wavefront of the probe beam is modulated by the DMD via binary holography. By switching the holograms stored in the DMD memory, the laser focus can be rapidly moved in space in a discrete fashion. The microscope system has a field of view of 65×130×155 µm3 in the x, y, and z axes, respectively; and a scanning speed of 8 kHz which is limited by the response time of the lock-in amplifier. To demonstrate the pump-probe system, we measured the ultrafast transient reflectivity of 2-D gold patterns on a silicon substrate and on silicon nitride cantilever beams. The results show an excellent signal-to-noise ratio and spatial-temporal resolution, as well as the 3-D random scanning capability. The new pump-probe microscope is a versatile instrument to characterize ultrafast 3-D phenomena with high spatial and temporal resolution, e.g., the propagation of localized surface plasmon resonance on curved surfaces.

Optical detection of ultrasound by lateral shearing interference of a transparent PDMS thin film.

A lateral shearing interferometric technique combined with an 11.6 µm polydimethylsiloxane (PDMS) transparent thin film is proposed and demonstrated for optical detection of ultrasound. We experimentally report the device change of reflectivity with pressure of 5.1×10-7 Pa-1, 9.5 times more sensitive than the critical-angle-based sensor, 31 times more sensitive than the surface-plasmon-based sensor, and comparable to the Fabry-Perot sensor. The objective-lens-based angle scanning characterization setup inspired from a laser scanning system allows direct comparison between the PDMS sensor and critical-angle-based sensor by adjusting the incident angle with a scanning mirror, thereby eliminating optical and electronics system dependence. The sensing element is easily fabricated through spin coating and the detection element incorporated into an existing optical system with minimum modification.

Common-path surface plasmon interferometer with radial polarization.

We present a common-path surface plasmon interferometer with radial polarization. We show how the V(z) effect, the output of the microscope versus defocus z, can be derived utilizing a radially polarized illumination and a virtual annulus. The measurement of the V(z) effect gives a strong signature of the surface plasmon propagation, which is functionally related to the material properties. We discuss the advantages of using radial polarization compared to linear polarization.

Hilbert transform-based single-shot plasmon microscopy.

The localized properties of surface plasmons (SPs) and surface waves can be measured with a modified confocal microscope. An interference signal arises from a locally generated reference close to normal incidence and the beam that forms the surface wave. A spatial light modulator can impose different phase shifts on the part of the incident light to recover the properties of the SP. We report a Hilbert transform method to recover the wavenumber with a single shot. The method is faster and potentially less expensive than previous approaches. The signal-to-noise ratio is equivalent to the phase-stepping method. The signal processing necessary to condition the signal is described.

Adjustable microscopic measurement of nanogap waveguide and plasmonic structures.

We investigate the performance of surface plasmon and Fabry-Perot modes formed between two closely spaced layers. The motivation for this study is twofold: first, to look for modes that may be excited at lower incident angles compared to the usual Kretschmann configuration with similar or superior refractive index responsivity and, second, to develop a simple and applicable method to study these structures over a wide range of separations without recourse to the construction of ad hoc structures. Using back focal plane observation and appropriate signal processing, we show results for the Otto configuration at visible wavelengths at a range of separations not reported hitherto. Moreover, we investigate a hybrid structure we call the Kretschmann-Otto configuration that gives modes that change continuously from a hybridized surface plasmon mode to a zero-order Fabry-Perot mode. The ability to change the separation to small gap distances enables us to examine the Fabry-Perot modes where we show that it has superior refractive index responsivity, by more than an order of magnitude, compared to the Kretschmann configuration.

Sensitive detection of voltage transients using differential intensity surface plasmon resonance system.

This paper describes theoretical and experimental study of the fundamentals of using surface plasmon resonance (SPR) for label-free detection of voltage. Plasmonic voltage sensing relies on the capacitive properties of metal-electrolyte interface that are governed by electrostatic interactions between charge carriers in both phases. Externally-applied voltage leads to changes in the free electron density in the surface of the metal, shifting the SPR position. The study shows the effects of the applied voltage on the shape of the SPR curve. It also provides a comparison between the theoretical and experimental response to the applied voltage. The response is presented in a universal term that can be used to assess the voltage sensitivity of different SPR instruments. Finally, it demonstrates the capacity of the SPR system in resolving dynamic voltage signals; a detection limit of 10mV with a temporal resolution of 5ms is achievable. These findings pave the way for the use of SPR systems in the detection of electrical activity of biological cells.

Single shot embedded surface plasmon microscopy with vortex illumination.

In previous work we demonstrated how a confocal microscope with a spatial light modulator in the back focal plane could perform accurate measurement of the k-vector of a propagating surface plasmon. This involved forming an embedded interferometer between light incident close to normal incidence (reference beam) and light incident at the angle to excite surface plasmons (sample beam). The signal from the interferometer was extracted by stepping the phase of the reference beam relative to the sample beam using a spatial light modulator; this requires at least 3 phase steps, which limits the speed of operation. To overcome this and extract the same information with a single measurement, we project an azimuthal varying phase between 0 and 2π in the central region of the back focal plane; corresponding to small angles of incidence. This projects a vortex beam as the reference, so that the phase of the reference beam varies with azimuthal angle. By extracting the interference signal from different portions of the reference beam, different phase steps between the reference and the sample are obtained, so all the values required for phase reconstruction can be extracted simultaneously. It is thus possible to obtain the same information with a single shot measurement, at each defocus position, without additional changes to the back focal plane illumination. Results are presented to show that the vortex illuminated sample provides similar results to the phase stepped version, whose values are, in turn, validated with ellipsometry and surface profilometry.

Grating-coupled Otto configuration for hybridized surface phonon polariton excitation for local refractive index sensitivity enhancement.

We demonstrate numerically through rigorous coupled wave analysis (RCWA) that replacing the prism in the Otto configuration with gratings enables us to excite and control different modes and field patterns of surface phonon polaritons (SPhPs) through the incident wavelength and height of the Otto spacing layer. This modified Otto configuration provides us the following multiple modes, namely, SPhP mode, Fabry-Pérot (FP) cavity resonance, dielectric waveguide grating resonance (DWGR) and hybridized between different combinations of the above mentioned modes. We show that this modified grating-coupled Otto configuration has a highly confined field pattern within the structure, making it more sensitive to local refractive index changes on the SiC surface. The hybridized surface phonon polariton modes also provide a stronger field enhancement compared to conventional pure mode excitation.

Live Imaging of Cellular Internalization of Single Colloidal Particle by Combined Label-Free and Fluorescence Total Internal Reflection Microscopy.

In this work we utilize the combination of label-free total internal reflection microscopy and total internal reflectance fluorescence (TIRM/TIRF) microscopy to achieve a simultaneous, live imaging of single, label-free colloidal particle endocytosis by individual cells. The TIRM arm of the microscope enables label free imaging of the colloid and cell membrane features, while the TIRF arm images the dynamics of fluorescent-labeled clathrin (protein involved in endocytosis via clathrin pathway), expressed in transfected 3T3 fibroblasts cells. Using a model polymeric colloid and cells with a fluorescently tagged clathrin endocytosis pathway, we demonstrate that wide field TIRM/TIRF coimaging enables live visualization of the process of colloidal particle interaction with the labeled cell structure, which is valuable for discerning the membrane events and route of colloid internalization by the cell. We further show that 500 nm in diameter model polystyrene colloid associates with clathrin, prior to and during its cellular internalization. This association is not apparent with larger, 1 µm in diameter colloids, indicating an upper particle size limit for clathrin-mediated endocytosis.

Effect of skin color on optical properties and the implications for medical optical technologies: a review.

Significance: Skin color affects light penetration leading to differences in its absorption and scattering properties. COVID-19 highlighted the importance of understanding of the interaction of light with different skin types, e.g., pulse oximetry (PO) unreliably determined oxygen saturation levels in people from Black and ethnic minority backgrounds. Furthermore, with increased use of other medical wearables using light to provide disease information and photodynamic therapies to treat skin cancers, a thorough understanding of the effect skin color has on light is important for reducing healthcare disparities. Aim: The aim of this work is to perform a thorough review on the effect of skin color on optical properties and the implication of variation on optical medical technologies. Approach: Published in vivo optical coefficients associated with different skin colors were collated and their effects on optical penetration depth and transport mean free path (TMFP) assessed. Results: Variation among reported values is significant. We show that absorption coefficients for dark skin are ∼6% to 74% greater than for light skin in the 400 to 1000 nm spectrum. Beyond 600 nm, the TMFP for light skin is greater than for dark skin. Maximum transmission for all skin types was beyond 940 nm in this spectrum. There are significant losses of light with increasing skin depth; in this spectrum, depending upon Fitzpatrick skin type (FST), on average 14% to 18% of light is lost by a depth of 0.1 mm compared with 90% to 97% of the remaining light being lost by a depth of 1.93 mm. Conclusions: Current published data suggest that at wavelengths beyond 940 nm light transmission is greatest for all FSTs. Data beyond 1000 nm are minimal and further study is required. It is possible that the amount of light transmitted through skin for all skin colors will converge with increasing wavelength enabling optical medical technologies to become independent of skin color.

Quantitative plasmonic measurements using embedded phase stepping confocal interferometry.

In previous publications [Opt. Express 20, 7388 (2012), Opt. Express 20, 28039 (2012)] we showed how a confocal configuration can form an surface plasmon microscope involving interference between a path involving the generation of surface plasmons and one involving a directly reflected beam. The relative phase of these contributions changes with axial scan position allowing the phase velocity of the surface plasmon to be measured. In this paper we extend the interferometer concept to produce an 'embedded' phase shifting interferometer, where we can control the phase between the reference and surface plasmon beams with a spatial light modulator. We demonstrate that this approach facilitates extraction of the amplitude and phase of the surface plasmon to measure of the phase velocity and the attenuation of the surface plasmons with greatly improved signal to noise compared to previous measurement approaches. We also show that reliable results are obtained over smaller axial scan ranges giving potentially superior lateral resolution.

Super-resolution imaging using proximity projection grating and structured light illumination.

This paper addresses optical super-resolution in the far field. We will describe the use of a novel optical component, which we call the proximity projection grating (PPG), that can provide different intensity patterns for sample illumination. These different illumination patterns allow the optical system to perform various modes of imaging, all are capable of resolution beyond the Abbe diffraction limit. Results will be shown to demonstrate the operations of some of these imaging modes. The potential of the PPG unit will also be discussed.

Wavelength-multiplexing phase-sensitive surface plasmon imaging sensor.

A wavelength-multiplexing phase-sensitive surface plasmon resonance (SPR) imaging sensor offering wide dynamic detection range and microarray capability is reported. Phase detection is accomplished by performing self-interference between the s- and p- polarizations within the signal beam. A liquid crystal tunable filter is used to sequentially select the SPR excitation wavelength from a white light source. This wavelength-multiplexing approach enables fast detection of the sensor's SPR phase response over a wide range of wavelengths, thereby covering literally any regions of interest within the SPR dip and thus maintaining the highest sensitivity point at all times. The phase-sensitive approach is particularly important for imaging SPR sensing applications because of its less stringent requirements for intensity signal-to-noise ratio, which also means the possibility of using uncooled modest resolution analog-to-digital conversion imaging devices. Experimental results demonstrate a resolution of 2.7×10(-7) RIU with a dynamic range of 0.0138 RIU.

Relevance and utility of the in-vivo and ex-vivo optical properties of the skin reported in the literature: a review [Invited].

Imaging non-invasively into the human body is currently limited by cost (MRI and CT scan), image resolution (ultrasound), exposure to ionising radiation (CT scan and X-ray), and the requirement for exogenous contrast agents (CT scan and PET scan). Optical imaging has the potential to overcome all these issues but is currently limited by imaging depth due to the scattering and absorption properties of human tissue. Skin is the first barrier encountered by light when imaging non-invasively, and therefore a clear understanding of the way that light interacts with skin is required for progress on optical medical imaging to be made. Here we present a thorough review of the optical properties of human skin measured in-vivo and compare these to the previously collated ex-vivo measurements. Both in-vivo and ex-vivo published data show high inter- and intra-publication variability making definitive answers regarding optical properties at given wavelengths challenging. Overall, variability is highest for ex-vivo absorption measurements with differences of up to 77-fold compared with 9.6-fold for the in-vivo absorption case. The impact of this variation on optical penetration depth and transport mean free path is presented and potential causes of these inconsistencies are discussed. We propose a set of experimental controls and reporting requirements for future measurements. We conclude that a robust in-vivo dataset, measured across a broad spectrum of wavelengths, is required for the development of future technologies that significantly increase the depth of optical imaging.

A total-internal-reflection-based Fabry-Pérot resonator for ultra-sensitive wideband ultrasound and photoacoustic applications.

In photoacoustic and ultrasound imaging, optical transducers offer a unique potential to provide higher responsivity, wider bandwidths, and greatly reduced electrical and acoustic impedance mismatch when compared with piezoelectric transducers. In this paper, we propose a total-internal-reflection-based Fabry-Pérot resonator composed of a 12-nm-thick gold layer and a dielectric resonant cavity. The resonator uses the same Kretschmann configuration as surface plasmon resonators (SPR). The resonators were analyzed both theoretically and experimentally. The experimental results were compared with those for an SPR for benchmarking. The 1.9-µm-thick-PMMA- and 3.4-µm-thick-PDMS-based resonators demonstrated responsivities of 3.6- and 30-fold improvements compared with the SPR, respectively. The measured bandwidths for the PMMA, PDMS devices are 110 MHz and 75 MHz, respectively. Single-shot sensitivity of 160 Pa is obtained for the PDMS device. The results indicate that, with the proposed resonator in imaging applications, sensitivity and the signal-to-noise ratio can be improved significantly without compromising the bandwidth.

Surface plasmon microscopic sensing with beam profile modulation.

Surface Plasmon microscopy enables measurement of local refractive index on a far finer scale than prism based systems. An interferometric or confocal system gives the so-called V(z) curve when the sample is scanned axially, which gives a measure of the surface plasmon propagation velocity. We show how a phase spatial light modulator (i) performs the necessary pupil function apodization (ii) imposes an angular varying phase shift that effectively changes sample defocus without any mechanical movement and (iii) changes the relative phase of the surface plasmon and reference beam to provide signal enhancement not possible with previous configurations.

Confocal surface plasmon microscopy with pupil function engineering.

Surface Plasmon microscopy can measure local changes of refractive index on the micron scale. Interferometric plasmon imaging delivers quantitative high spatial resolution sensitive to refractive index. In addition the so called V(z) method allows image contrast to be controlled by varying the sample defocus without substantially degrading spatial resolution. Here, we show how a confocal system provides a simpler and more stable alternative. This system, however, places greater demands on the dynamic range of the system. We therefore use a spatial light modulator to engineer the microscope pupil function to suppress light that does not contribute to the signal.

Determination of crystallographic orientation of large grain metals with surface acoustic waves.

A previously described laser ultrasonic technique known as spatially resolved acoustic spectroscopy (SRAS) can be used to image surface microstructure, using the local surface acoustic wave (SAW) velocity as a contrast mechanism. It is shown here that measuring the SAW velocity in multiple directions can be used to determine the crystallographic orientation of grains. The orientations are determined by fitting experimentally measured velocities to theoretical velocities. Using this technique the orientations of 12 nickel and 3 aluminum single crystal samples have been measured, and these are compared with x-ray Laue backreflection (LBR) measurements with good agreement. The root mean square difference between SRAS and LBR measurements in terms of an R-value is less than 4.1°. The influence of systematic errors in the SAW velocity determination due to instrument miscalibration, which affects the accurate determination of the planes, is discussed. SRAS has great potential for complementary measurements or even for replacing established orientation determination and imaging techniques.

Polarization modulation thermal lens microscopy for imaging the orientation of non-spherical nanoparticles.

In this paper a far field optical technique we call polarization modulation thermal lens microscopy (PM-TLM) is used for imaging the orientation and dichroism of non-spherical nanoparticles. In PM-TLM, the polarization state of a pump beam is periodically modulated which in turn causes morphology related intensity fluctuations in a continuous probe beam, thus allowing high signal to noise ratio detection with using lock-in amplification. Since PM-TLM uses nanoparticle absorption as the contrast mechanism, it may be used to detect and image nanoparticles of far smaller dimensions than can be observed by conventional dark field optical microscopy. The technique, its implementation and experiment results are presented.