Reversible inter-degree-of-freedom optical-coherence conversion via entropy swapping.

The entropy associated with an optical field quantifies the field fluctuations and thus its coherence. Any binary optical degree-of-freedom (DoF) - such as polarization or the field at a pair of points in space - can each carry up to one bit of entropy. We demonstrate here that entropy can be reversibly swapped between different DoFs, such that coherence is converted back and forth between them without loss of energy. Specifically, starting with a spatially coherent but unpolarized field carrying one bit of entropy, we unitarily convert the coherence from the spatial DoF to polarization to produce a spatially incoherent but polarized field by swapping the entropy between the two DoFs. Next, we implement the inverse unitary operator, thus converting the coherence back to yield once again a spatially coherent yet unpolarized field. We exploit the intermediate stage between the two coherence conversions - where the spatial coherence has been converted to the polarization DoF - to verify that the field has become immune to the deleterious impact of spatial phase scrambling. Maximizing the spatial entropy protects the spatial DoF by preventing it from taking on any additional fluctuations. After the second coherence conversion, spatial coherence is readily retrieved, and the effect of spatial phase scrambling circumvented.

Demonstration of speckle resistance using space-time light sheets.

The capacity of self-healing fields to reconstruct after passing through scattering media may prove useful in reducing speckle formation. Here, we study the speckle response of the space-time (ST) light sheet compared to a Gaussian wave packet, Airy beam, and Bessel Gauss beam. We find that the Pearson's correlation coefficient for the ST light sheet is 50%, 48% and 40% larger than that of the Gaussian, Airy beam and Bessel Gauss beams, respectively, demonstrating a strong correlation to an input beam that has not been speckled. These results suggest that the ST light sheet exhibits considerable resistance to speckle generation. We also investigate the speckle response of the ST light sheet at its second-harmonic frequency and observe a mean Pearson's correlation coefficient close to 0.6, comparable to the second-harmonic Bessel Gauss beam, and 2.8 × the value obtained for the second-harmonic Gaussian beam. Our results lend themselves to a variety of applications including bioimaging, communications, and optical tweezers.

Space-time vector light sheets.

We introduce the space-time (ST) vector light sheet. This unique one-dimensional ST wave packet is characterized by classical entanglement (CE), a correlation between at least two non-separable intrinsic degrees-of-freedom (DoFs), which in this case are the spatiotemporal DoFs in parallel with the spatial-polarization DoFs. We experimentally confirm that the ST vector light sheet maintains the intrinsic features of the uniformly polarized ST light sheet, such as near-diffraction-free propagation and self-healing, while also maintaining the intrinsic polarization structure of common vector beams, such as those that are radially polarized and azimuthally polarized. We also show that the vector beam structure of the ST vector light sheet is maintained in the subluminal and superluminal regimes.

Multiphoton Microscopy for the Characterization of Cellular Behavior on Naturally Derived Polysaccharide Tissue Constructs With Irregular Surfaces for the Development of Platform Biomaterials.

Over the past decade, the use of polymers as platform materials for biomedical applications including tissue engineering has been of rising interest. Recently, the use of naturally derived polysaccharides as 3-D scaffolds for tissue regeneration has shown promising material characteristics; however, due to complexities in composition, morphology, and optical properties, adequate spatial and temporal characterization of cellular behavior in these materials is lacking. Multiphoton microscopy has emerged as a viable tool for performing such quantification by permitting greater imaging depth while simultaneously minimizing un-favorable scattering and producing high-resolution optical cross sections for non-invasive analysis. Here we describe a method using endogenous contrast of cellulose nanofibers (CNF) using Second Harmonic Generation (SHG), combined with 2-photon fluorescence of Cell Tracker Orange for spatial and longitudinal imaging of cellular proliferation. Cell Tracker Orange is an ideal fluorophore to avoid the broad CNF autofluorescence allowing for segmentation of cells using a semi-automatic routine. Individual cells were identified using centroid locations for 3D cell proliferation. Overall, the methods presented are viable for investigation of cellular interactions with polysaccharide candidate biomaterials.

Global analysis and Decay Associated Images (DAI) derived from Fluorescence Lifetime Imaging Microscopy (FLIM).

The extraction of fluorophore lifetimes in a biological sample provides useful information about the probe environment that is not readily available from fluorescence intensity alone. Cell membrane potential, pH, concentration of oxygen ([O2]), calcium ([Ca2+]), NADH and other ions and metabolites are all regularly measured by lifetime-based techniques. These measurements provide invaluable knowledge about cell homeostasis, metabolism and communication with the cell environment. Fluorescence lifetime imaging microscopy (FLIM) produces spatial maps with time-correlated single-photon counting (TCSPC) histograms collected and analyzed at each pixel, but traditional TCSPC analysis is often hampered by the low number of photons that can reasonably be collected while maintaining high spatial resolution. More important, traditional analysis fails to employ the spatial linkages within the image. Here, we present a different approach, where we work under the assumption that mixtures of a global set of lifetimes (often only 2 or 3) can describe the entire image. We determine these lifetime components by globally fitting precise decays aggregated over large spatial regions of interest, and then we perform a pixel-by-pixel calculation of decay amplitudes (via simple linear algebra applied to coarser time-windows). This yields accurate amplitude images (Decay Associate Images, DAI) that contain stoichiometric information about the underlying mixtures while retaining single pixel resolution. We collected FLIM data of dye mixtures and bacteria expressing fluorescent proteins with a two-photon microscope system equipped with a commercial single-photon counting card, and we used these data to benchmark the gDAI program.