Browsing by Subject "Nonlinear microscopy"

Imaging based on nonlinear processes takes advantage of the localized excitation to achieve high spatial resolution, optical sectioning, and deeper penetration in highly scattering media. However, the use of nonlinear contrast for imaging has conventionally been limited to processes that create light of wavelengths that are different from the wavelengths used for excitation. Intrinsic nonlinear contrasts that do not generate light at distinct wavelengths are generally difficult to measure because of the overwhelming background from the excitation light. This dissertation focuses on extension of nonlinear microscopy to these new intrinsic processes by using femtosecond pulse shaping to encode the nonlinear information as new frequency components in the spectrum. We will present a pump-probe microscopy technique based on pulse train shaping technology to sensitively access nonlinear transient absorption or gain processes. This technique has recently been used to uniquely identify a variety of biological pigments with high spatial resolution. Here, we extend this technique to image and characterize several inorganic and organic pigments used in historical artwork. We also present a spectral reshaping technique based on individual femtosecond pulse shaping to sensitively access nonlinear refractive contrasts in scattering media. We will describe an extension of this technique to utilize two distinct wavelengths and discuss its application in biological imaging. This two-color implementation would allow the extension of widely employed phase contrast to the nonlinear regime.

First mined in Afghanistan nearly 6,000 years ago, lapis lazuli is a blue pigment also known as ultramarine, a material that was highly prized (and corresponding highly priced) by Western painters during the Middle Ages and the Italian Renaissance. Both the mineral lapis lazuli and the synthetic ultramarine can undergo degradation in paintings and other works of cultural heritage, which presents challenges to the preservations of these works. Due to the limitations of many modern analytical techniques, art conservators and conservation scientists often still need to remove a sample of paint in order to understand the layering of pigments in a painting. Femtosecond transient absorption (also called pump-probe) spectroscopy and imaging are here used to explore the effects of depth, polarization, and power on the ultrafast excited state dynamics of ultramarines both natural and synthetic, in order to further understand the photophysics and potential photo-degradation pathways of ultramarine pigments in paintings. Both lapis lazuli and synthetic ultramarine undergo identical forms of photo-induced transformation in the context of these experiments, where it appears that either the lazurite chromophore or the sodalite cage structure of ultramarine is destroyed.

Nonlinear optical microscopy serves as a great tool for biomedical imaging due to its high resolution, deep penetration, inherent three dimensional optical sectioning capabilities and superior performance in scattering media. Conventional nonlinear optical microscopy techniques, e.g. two photon fluorescence and second harmonic generation, are based on detecting a small light signal emitted at a new wavelength that is well separated from the excitation light. However, there are also many other nonlinear processes, such as two-photon absorption and self-phase modulation, that do not generate light at new wavelengths and that have not been extensively explored for imaging. This dissertation extends the accessible mechanisms for contrast to the later nonlinear optical processes by combining femtosecond laser pulse shaping and homodyne detection. We developed a rapid pulse shaper with a relatively simple and compact instrument design that modifies the spectrum of individual laser pulses from an 80 MHz mode-locked laser. The pulse shaper enables simultaneous two-photon absorption and self-phase modulation imaging of various nanoparticles in-vitro with high sensitivity. We also applied this imaging technique to study the nonlinear optical response in graphene. Because our technology detects the nonlinear signature encoded within the laser pulse itself, we achieve intrinsic contrast of biological and non-biological samples in highly scattering media. These capabilities have significant implications in biomedical imaging and nanophotonics.

The materials and working method of a painting can reveal important information about our cultural history, as well as lend the conservator the necessary knowledge for treatment options. The removal of a cross-section sample reveals the three-dimensional (3d) structure of the painting and can be used to identify materials. However, cross-section samples are destructive and provide only local information. Nonlinear optical ultrafast pump-probe microscopy, originally developed for biomedical imaging, can provide high resolution 3d images with chemical contrast. In this dissertation, I adapt pump-probe microscopy to multiple materials and applications in cultural heritage research. Pump-probe dynamics were found to be sensitive to the ratio of the two chromophores present in the precious blue pigment lapis lazuli and its synthetic analogs, ultramarines blue and violet. Virtual pump-probe cross-sections were combined with nonlinear fluorescence contrast to study differences between the interactions of paper supports with inorganic crystalline pigments and organic dyes. Multiple early Italian paintings (The Crucifixion by Puccio Capanna, The Martyrdom of St. Alexander and The Body of Christ Supported by Angels attributed to Lorenzo Lotto) were imaged in-situ, in conjunction with traditional conservation science methods, as a part of a technical case study. Thus, pump-probe microscopy offers an important new tool for gaining fundamental insights into our cultural heritage.