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Spectral self-interference microscopy (SSM) is a fluorescence imaging technique developed in my former group at Boston University. In SSM an object is stained with fluorescent molecules and placed above a mirror. The fluorophores are optically excited and emit light toward the observer and toward the mirror. The resulting effect is directly analogous to soap bubble interference. The light coming out of the sample displays spectral patterns which can be measured with a spectrometer. By analyzing these patterns we are able to infer the fluorophore-to-mirror distance with nanometer precision. Researchers are currently making great strides in the understanding of molecular and sub-cellular biological processes. Fluorescence microscopy is a very important tool in this effort, as fluorescent molecules can be attached to prescribed biological structures with great specificity and imaged in vivo. SSM has the potential to enable important new fluorescence studies, as it allows nanoscale depth localization of the fluorescent dyes. For example, SSM was demonstrated in the study of DNA confirmation in Moiseev et al., PNAS, 2006. I was brought into the SSM project soon after the technique was conceived, and contributed to the first journal-published description of SSM. My doctoral work focused on providing a full mathematical description of the capabilities (and limitations) of SSM. This work also led to the recognition of a connection between SSM and 4Pi microscopy, a technique developed in the group of Dr. Stefan Hell. A full description of the capabilites of SSM in noisy imaging environments has also been published and includes the use of an evanescent field to excite the fluorescent object.
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Interferometric Synthetic Aperture Microscopy (ISAM) is an optical imaging technique developed and demonstrated by Ralston, Marks, Carney, and Boppart. Rather than imaging a fluorescent sample, ISAM measures light scattered from an unstained object. ISAM is based on Optical Coherence Tomography (OCT), a maturing noninvasive technique that has already found application, particularly in opthamology. In addition, the Boppart group is actively pursuing the application of OCT and ISAM to other biomedical tasks, such as breast tissue imaging.
OCT and ISAM both rely on interferometric detection of scattered light. A broadband source is used and the resulting data is a function of two spatial variables and the wavelength - (x,y,λ). With the correct processing the data can be used to infer object structure in three spatial dimensions - (x,y,z). In OCT the image is reconstructed only where the probing light is in focus. The achievement of ISAM is to recognize that light outside of the focus can be computationally focused after data acquisition. This greatly improves the depth-of-field available. As in OCT, the processing used in ISAM requires interferometric detection of data, so that phase information is available. As the name of this technology indicates, ISAM has strong commonalities with Synthetic Aperture Radar (SAR). OCT can be viewed as analogous to standard radar.
I joined the ISAM research effort after the first physical demonstrations. My work includes a full electromagnetic high-aperture model for ISAM, which generalizes previous scalar results. The electromagnetic model clarified the mechanisms behind ISAM - in particular it was shown that the stationary-phase approximation should be used to derive the ISAM processing algorithm for scatterers that lie out of focus. Despite the nonparaxial and vectorial nature of the model, it was shown that the main action of the ISAM processing can be reduced to a simple Fourier-domain data warping. In addition, I was involved in demonstrating that ISAM implicitly mitigates the autocorrelation artifact produced by frequency-domain data collection.
Optical fields are subject to random fluctuations and can therefore be considered statistically. The study of optical-field statistics is usually known as "coherence theory." Many optical fields are statistically stationary which means that the statistics do not vary with time. The assumption of stationarity allows the invocation of the celebrated Weiner-Khintchine theorem. This theorem states that each frequency component (i.e. each color or wavelength) is independent of all other frequency components. This useful result means that an optical field can be analyzed frequency-by-frequency and the results incoherently summed to get the total field. Many stationary fields are also ergodic, which means that the field statistics can be estimated from physical measurements. The use of pulsed lasers is becoming more prevalent in modern optical applications. Fields from pulsed lasers have a repeating temporal structure and are therefore nonstationary. This introduces correlations in the spectrum (as shown in the diagram above) and necessitates the use of a more general coherence theory. In addition, ergodicity is no longer applicable and care needs to be taken with the interpretation of optical measurements. I have applied the theory of cyclostationary random processes to optical fields. A cyclostationary field has statistics that repeat periodically - a model applicable to pulsed lasers. Cyclostationarity and cycloergodicity allow the construction of a coherence theory for pulsed fields that can be used to guide the interpretation of physical measurements. I have shown how standard ultrafast pulse measurement instruments can be used in a many-pulse regime (generally, the standard use is to measure only a single pulse) and the resulting data used to infer nonstationary statistics of the field. This indicates a direction toward the comprehensive characterization of pulsed laser stability. In addition to contributing to nonstationary coherence theory, I have also developed numerical space-time simulators for stationary fields. | ||
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Like ISAM, synthetic aperture sonar (SAS) and synthetic aperture radar are coherent imaging techniques that use image reconstruction to synthesize an image. As part of the masters program at the University of Arizona I performed research into the effects of multipath propagation in SAS imaging. My work showed that the SAS processing localizes the desirable line-of-sight signal but leaves the multipath artifacts blurred.
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Comment / Questions to David Lytle, lytle@uiuc.edu |
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