Eric D. Diebold

Digitally synthesized beat frequency multiplexing for sub-millisecond fluorescence microscopy

A confocal fluorescence microscopy scheme that maps the image to the radiofrequency spectrum by beating together two optical fields offers enhanced read-out speeds at kilohertz frame rates. It provides a new way for observing dynamic phenomena in cells.

Giant tunable optical dispersion using chromo-modal excitation of a multimode waveguide

The ability to control chromatic dispersion is paramount in applications where the optical pulsewidth is critical, such as chirped pulse amplification and fiber optic communications. Typically, devices used to generate large amounts (>100 ps/nm) of chromatic dispersion are based on diffraction gratings, chirped fiber Bragg gratings, or dispersion compensating fiber. Unfortunately, these dispersive elements suffer from one or more of the following restrictions: (i) limited operational bandwidth, (ii) limited total dispersion, (iii) low peak power handling, or (iv) large spatial footprint. Here, we introduce a new type of tunable dispersive device, which overcomes these limitations by leveraging the large modal dispersion of a multimode waveguide in combination with the angular dispersion of diffraction gratings to create chromatic dispersion. We characterize the devices dispersion, and demonstrate its ability to stretch a sub-picosecond optical pulse to nearly 2 nanoseconds in 20 meters of multimode optical fiber. Using this device, we also demonstrate single-shot, time-wavelength atomic absorption spectroscopy at a repetition rate of 90.8 MHz.

Tailoring the biodegradability of porous silicon nanoparticles

Porous silicon nanoparticles (PSiNPs) are attractive carriers for targeted drug delivery in nanomedicine. For in vivo applications, the biodegradation property of PSiNPs provides a pathway for their safe clearance from the body. Particles sizes of 80-120 nm are of particular interest as they are important for cellular applications, such as drug delivery for cancer therapy, because these nanoparticles can take advantage of the enhanced permeability and retention effect to deliver drug preferentially to tumors with leaky vasculature, yet large enough to avoid renal clearance. However, the biodegradability rate of such particles is often too fast, which limits particle half-life and potentially reduces their in vivo delivery efficiency. In this work, we focus on the degradation of nanoscale particles and study the effect of both thermal oxidation and silica coating on the stability of PSiNPs in phosphate buffered saline solution (a close mimic of a basic biological fluid). Using thermal oxidation, the half-life of PSiNPs can be varied from 10 min up to 3 h. Using silica coating, the half-life can be extended further to 8 h. The particles produced using both these techniques can be functionalized using standard silica surface chemistries developed for applications in drug delivery.

Active THz medical imaging using broadband direct detection

Research in THz imaging is generally focused on three primary application areas: medical, security, and nondestructive evaluation (NDE). While work in THz security imaging and personnel screening is populated by a number of different active and passive system architectures, research in medical imaging in is generally performed with THz time-domain systems. These systems typically employ photoconductive or electro-optic source/detector pairs and can acquire depth resolved data or spectrally resolved pixels by synchronously sampling the electric field of the transmitted/reflected waveform. While time-domain is a very powerful scientific technique, results reported in the literature suggest that desired THz contrast in medical imaging may not require the volume of data accessible from time-resolved measurements and that a simpler direct detection, active technique may be sufficient for specific applications. In this talk we discuss an active direct detection reflectometer system architecture operating at a center frequency of ~ 525 GHz that uses a photoconductive source and schottky diode detector. This design takes advantage or radar-like pulse rectification and novel reflective optical design to achieve high target imaging contrast with significant potential for high speed acquisition time. Results in spatially resolved hydration mapping of burn wounds are presented and future outlooks discussed.

Spectrally encoded angular light scattering

The angular light scattering profile of microscopic particles significantly depends on their morphological parameters, such as size and shape. This dependency is widely used in state-of-the-art flow cytometry methods for particle classification. We introduce a new spectrally encoded angular light scattering method, with potential application in scanning flow cytometry. We show that a one-to-one wavelength-to-angle mapping enables the measurement of the angular dependence of scattered light from microscopic particles over a wide dynamic range. Improvement in dynamic range is obtained by equalizing the angular dependence of scattering via wavelength equalization. Continuous angular spectrum is obtained without mechanical scanning enabling single-shot measurement. Using this information, particle morphology can be determined with improved accuracy. We derive and experimentally verify an analytic wavelength-to-angle mapping model, facilitating rapid data processing. As a proof of concept, we demonstrate the methods capability of distinguishing differently sized polystyrene beads. The combination of this technique with time-stretch dispersive Fourier transform offers real-time and high-throughput (high frame rate) measurements and renders the method suitable for integration in standard flow cytometers.

Real-time wavelength and bandwidth-independent optical integrator based on modal dispersion.

High-throughput real-time optical integrators are of great importance for applications that require ultrafast optical information processing, such as real-time phase reconstruction of ultrashort optical pulses. In many of these applications, integration of wide optical bandwidth signals is required. Unfortunately, conventional all-optical integrators based on passive devices are usually sensitive to the wavelength and bandwidth of the optical carrier. Here, we propose and demonstrate a passive all-optical intensity integrator whose operation is independent of the optical signal wavelength and bandwidth. The integrator is implemented based on modal dispersion in a multimode waveguide. By controlling the launch conditions of the input beam, the device produces a rectangular temporal impulse response. Consequently, a temporal intensity integration of an arbitrary optical waveform input is performed within the rectangular time window. The key advantage of this device is that the integration operation can be performed independent of the input signal wavelength and optical carrier bandwidth. This is preferred in many applications where optical signals of different wavelengths are involved. Moreover, thanks to the use of a relatively short length of multimode waveguide, lower system latency is achieved compared to the systems using long dispersive fibers. To illustrate the versatility of the optical integrator, we demonstrate temporal intensity integration of optical waveforms with different wavelengths and optical carrier bandwidths. Finally, we use this device to perform high-throughput, single-shot, real-time optical phase reconstruction of phase-modulated signals at telecommunications bit rates.

Digitally synthesized beat frequency-multiplexed fluorescence lifetime spectroscopy

Frequency domain fluorescence lifetime imaging is a powerful technique that enables the observation of subtle changes in the molecular environment of a fluorescent probe. This technique works by measuring the phase delay between the optical emission and excitation of fluorophores as a function of modulation frequency. However, high-resolution measurements are time consuming, as the excitation modulation frequency must be swept, and faster low-resolution measurements at a single frequency are prone to large errors. Here, we present a low cost optical system for applications in real-time confocal lifetime imaging, which measures the phase vs. frequency spectrum without sweeping. Deemed Lifetime Imaging using Frequency-multiplexed Excitation (LIFE), this technique uses a digitally-synthesized radio frequency comb to drive an acousto-optic deflector, operated in a cats-eye configuration, to produce a single laser excitation beam modulated at multiple beat frequencies. We demonstrate simultaneous fluorescence lifetime measurements at 10 frequencies over a bandwidth of 48 MHz, enabling high speed frequency domain lifetime analysis of single- and multi-component sample mixtures.

Single-Cell Analysis of Morphological and Metabolic Heterogeneity in Euglena gracilis by Fluorescence-Imaging Flow Cytometry

Microalgal biofuels and biomass have ecofriendly advantages as feedstocks. Improved understanding and utilization of microalgae require large-scale analysis of the morphological and metabolic heterogeneity within populations. Here, with Euglena gracilis as a model microalgal species, we evaluate how fluorescence- and brightfield-derived-image-based descriptors vary during environmental stress at the single-cell level. This is achieved with a new multiparameter fluorescence-imaging cytometric technique that allows the assaying of thousands of cells per experiment. We track morphological changes, including the intensity and distribution of intracellular lipid droplets, and pigment autofluorescence. The combined fluorescence-morphological analysis identifies new metrics not accessible with traditional flow cytometry, including the lipid-to-cell-area ratio (LCAR), which shows promise as an indicator of oil productivity per biomass. Single-cell metrics of lipid productivity were highly correlated ( R2 > 0.90, p < 0.005) with bulk oil extraction. Such chemomorphological atlases of algal species can help optimize growth conditions and selection approaches for large-scale biomass production.

Digitally-synthesized beat frequency multiplexing for sub-millisecond fluorescence microscopy

Radiofrequency beating of digitally synthesized optical fields extends the time resolution of fluorescence microscopy and enhances the throughput of flow imaging. We demonstrate fluorescence microscopy with pixel readout rates 30x faster than a modern EMCCD.

Radiofrequency encoded angular-resolved light scattering

The sensitive, specific, and label-free classification of microscopic cells and organisms is one of the outstanding problems in biology. Today, instruments such as the flow cytometer use a combination of light scatter measurements at two distinct angles to infer the size and internal complexity of cells at rates of more than 10 000/s. However, by examining the entire angular light scattering spectrum, it is possible to classify cells with higher resolution and specificity. Current approaches to performing these angular spectrum measurements all have significant throughput limitations, making them incompatible with other state-of-the-art flow cytometers. Here, we introduce a method for performing complete angular scattering spectrum measurements at high throughput combining techniques from the field of scattering flow-cytometry and radiofrequency communications. Termed Radiofrequency Encoded Angular-resolved Light Scattering, this technique multiplexes angular light scattering in the radiofrequency domain,...