Supplementary document for Fisher Information and Cram\'er-Rao Lower Bound in Single Pixel Localization Microscopy with Spatio-temporally Modulated Illumination - 6135085.pdf

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Supplementary document for Super resolution computational saturated absorption microscopy - 6449981.pdf

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Supplementary document for Nearly degenerate two color impulsive coherent Raman hyperspectral imaging - 6044818.pdf

Latex for a supplemental document with expanded experimental method

Super resolution computational saturated absorption microscopy

Imaging beyond the diffraction limit barrier has attracted wide attention due to the ability to resolve image features that were previously hidden. Of the various super-resolution microscopy techniques available, a particularly simple method called saturated excitation microscopy (SAX) requires only a simple modification of a laser scanning microscope where the illumination beam power is sinusoidally modulated and driven into saturation. SAX images are extracted from harmonics of the modulation frequency and exhibit improved spatial resolution. Unfortunately, this elegant strategy is hindered by the incursion of shot noise that prevents high resolution imaging in many realistic scenarios. Here, we demonstrate a new technique for super resolution imaging that we call computational saturated absorption (CSA) in which a joint deconvolution is applied to a set of images with diversity in spatial frequency support among the point spread functions used in the image formation with saturated laser scanning fluorescence microscope. CSA microscopy allows access to the high spatial frequency diversity in a set of saturated effective point spread functions, while avoiding image degradation from shot noise

Hyperspectral acquisition with ScanImage at the single pixel level: Application to time domain coherent Raman imaging

We present a comprehensive strategy and its practical implementation using the commercial ScanImage software platform to perform hyperspectral point scanning microscopy when a fast time dependent signal varies at each pixel level. In the proposed acquisition scheme the scan along the X axis is slowed down while the data acquisition is maintained at high pace to enable the rapid acquisition of the time dependent signal at each pixel level. The ScanImage generated raw 2D images have a very asymmetric aspect ratio between X and Y, the X axis encoding both for space and time acquisition. The results are X axis macro-pixel where the associated time depend signal is sampled therefore providing an hyperspectral information. We exemplified the proposed hyperspectral scheme in the context of time domain coherent Raman imaging where a pump pulse impulsively excites molecular vibrations that are subsequently probed by a time delayed probe pulse. In this case the time dependent signal is a fast acousto-optics delay line that can scan a delay of 4.5ps in 25$\mu$s, at each pixel level. We this acquisition scheme we demonstrate ultra-fast hyperspectral vibrational imaging in the low frequency range [10$cm^{-1}$, 150 $cm^{-1}$] over a 500 $\mu m$ field of view in 14ms (7 frames/s). The proposed acquisition scheme can be readily extended to other applications requiring to acquired a fast evolving signal at each pixel level

Synthetic aperture holographic third harmonic generation microscopy

Third harmonic generation (THG) provides a valuable, label-free approach to imaging biological systems. To date, THG microscopy has been performed using point scanning methods that rely on intensity measurements lacking phase information of the complex field. We report the first demonstration of THG holographic microscopy and the reconstruction of the complex THG signal field with spatial synthetic aperture imaging. Phase distortions arising from measurement-to-measurement fluctuations and imaging components cause optical aberrations in the reconstructed THG field. We have developed an aberration-correction algorithm that estimates and corrects for these phase distortions to reconstruct the spatial synthetic aperture THG field without optical aberrations