Multicolored Stain-Free Histopathology with Coherent Raman Imaging

Conventional histopathology with hematoxylin & eosin (H&E) has been the gold standard for histopathological diagnosis of a wide range of diseases. However, it is not performed in vivo and requires thin tissue sections obtained after tissue biopsy, which carries risk, particularly in the central nervous system. Here we describe the development of an alternative, multicolored way to visualize tissue in real-time through the use of coherent Raman imaging (CRI), without the use of dyes. CRI relies on intrinsic chemical contrast based on vibrational properties of molecules and intrinsic optical sectioning by nonlinear excitation. We demonstrate that multicolor images originating from $CH_2$ and $CH_3$ vibrations of lipids and protein, as well as two-photon absorption of hemoglobin, can be obtained with subcellular resolution from fresh tissue. These stain-free histopathological images show resolutions similar to those obtained by conventional techniques, but do not require tissue fixation, sectioning or staining of the tissue analyzed.Chemistry and Chemical Biolog

Integrated liquid-core optical fibers --- ultra-efficient nonlinear liquid photonics

We have developed a novel integrated platform for liquid photonics based on liquid core optical fiber (LCOF). The platform is created by fusion splicing liquid core optical fiber to standard single-mode optical fiber making it fully integrated and practical - a major challenge that has greatly hindered progress in liquid-photonic applications. As an example, we report here the realization of ultralow threshold Raman generation using an integrated CS2 filled LCOF pumped with sub-nanosecond pulses at 1064nm and 532nm. The measured energy threshold for the Stokes generation is ~ 1nJ, about three orders of magnitude lower than previously reported values in the literature for hydrogen gas. The integrated LCOF platform opens up new possibilities for ultralow power nonlinear optics such as efficient white light generation for displays, mid-IR generation, slow light generation, parametric amplification, all-optical switching and wavelength conversion using liquids that have orders of magnitude larger optical nonlinearities compared with silica glass.Comment: 4 pages, 3 figure

The Borexino detector at the Laboratori Nazionali del Gran Sasso

Borexino, a large volume detector for low energy neutrino spectroscopy, is currently running underground at the Laboratori Nazionali del Gran Sasso, Italy. The main goal of the experiment is the real-time measurement of sub MeV solar neutrinos, and particularly of the mono energetic (862 keV) Be7 electron capture neutrinos, via neutrino-electron scattering in an ultra-pure liquid scintillator. This paper is mostly devoted to the description of the detector structure, the photomultipliers, the electronics, and the trigger and calibration systems. The real performance of the detector, which always meets, and sometimes exceeds, design expectations, is also shown. Some important aspects of the Borexino project, i.e. the fluid handling plants, the purification techniques and the filling procedures, are not covered in this paper and are, or will be, published elsewhere (see Introduction and Bibliography).Comment: 37 pages, 43 figures, to be submitted to NI

Ecological variation drives morphological differentiation in a highly social vertebrate

Animals may respond to ecological heterogeneity by genetic differentiation or phenotypic plasticity. Responses of organisms to their ecology can include adaptation at various levels of organization, including morphology, behaviour and social structure. Adaptations at one level might constrain or enhance adaptations on other levels, which highlights the importance of understanding their interactions. In highly social animals, understanding the influence of their ecological niche on the evolution and maintenance of complex social organization poses an intriguing challenge. Predation risk and habitat characteristics determine social structure in the cooperatively breeding cichlid Neolamprologus pulcher. Here we examine how varying combinations of these ecological factors across eight distinct populations influence morphological differentiation. We investigated the relationship between body shape and ecological parameters for 137 wild-caught individuals from eight distinct populations. Furthermore, we examined the genetic structure and differentiation among these populations using microsatellites. Finally, to disentangle heritable from plastic responses, we raised two successive generations from six populations in the laboratory under common garden conditions and screened 188 individuals for morphological differentiation. We found that body shape of N. pulcher strongly correlates with the measured ecological parameters. Low predation risk, low habitat structure and small shelter size favoured shallow bodies, whereas at the opposite end of these environmental gradients deep body shapes prevail. These consistent morphological differences persisted over two laboratory-reared generations, revealing a heritable basis. In contrast to the significant effect of local ecology on morphological differentiation between populations, both geographical and genetic distance had little explanatory power, suggesting that morphological differentiation between populations is not a simple by-product of genetic isolation. Remarkably, the largest difference in body depth emerged between the two populations located closest to each other, but differing strongly in their ecological niche. These results highlight that morphological variation is a key component of local adaptation in neighbouring populations of a highly social species. Such morphological differentiation has the potential to influence individual cooperative behaviour, which will eventually feed back on group structure and mediate the evolution and maintenance of complex social systems

New limits on nucleon decays into invisible channels with the BOREXINO Counting Test Facility

The results of background measurements with the second version of the BOREXINO Counting Test Facility (CTF-II), installed in the Gran Sasso Underground Laboratory, were used to obtain limits on the instability of nucleons, bounded in nuclei, for decays into invisible channels ($inv$): disappearance, decays to neutrinos, etc. The approach consisted of a search for decays of unstable nuclides resulting from $N$ and $NN$ decays of parents $^{12}$C, $^{13}$C and $^{16}$O nuclei in the liquid scintillator and the water shield of the CTF. Due to the extremely low background and the large mass (4.2 ton) of the CTF detector, the most stringent (or competitive) up-to-date experimental bounds have been established: $\tau(n \to inv) > 1.8 \cdot 10^{25}$ y, $\tau(p \to inv) > 1.1 \cdot 10^{26}$ y, $\tau(nn \to inv) > 4.9 \cdot 10^{25}$ y and $\tau(pp \to inv) > 5.0 \cdot 10^{25}$ y, all at 90% C.L.Comment: 22 pages, 3 figures,submitted to Phys.Lett.

New results on solar neutrino fluxes from 192 days of Borexino data

We report the direct measurement of the ^7Be solar neutrino signal rate performed with the Borexino detector at the Laboratori Nazionali del Gran Sasso. The interaction rate of the 0.862 MeV ^7Be neutrinos is 49+-3(stat)+-4(syst) counts/(day * 100ton). The hypothesis of no oscillation for ^7Be solar neutrinos is inconsistent with our measurement at the 4sigma level. Our result is the first direct measurement of the survival probability for solar nu_e in the transition region between matter-enhanced and vacuum-driven oscillations. The measurement improves the experimental determination of the flux of ^7Be, pp, and CNO solar nu_e, and the limit on the magnetic moment of neutrinos

Rapid, Label-Free Detection of Brain Tumors with Stimulated Raman Scattering Microscopy

Surgery is an essential component in the treatment of brain tumors. However, delineating tumor from normal brain remains a major challenge. We describe the use of stimulated Raman scattering (SRS) microscopy for differentiating healthy human and mouse brain tissue from tumor-infiltrated brain based on histoarchitectural and biochemical differences. Unlike traditional histopathology, SRS is a label-free technique that can be rapidly performed in situ. SRS microscopy was able to differentiate tumor from nonneoplastic tissue in an infiltrative human glioblastoma xenograft mouse model based on their different Raman spectra. We further demonstrated a correlation between SRS and hematoxylin and eosin microscopy for detection of glioma infiltration (κ = 0.98). Finally, we applied SRS microscopy in vivo in mice during surgery to reveal tumor margins that were undetectable under standard operative conditions. By providing rapid intraoperative assessment of brain tissue, SRS microscopy may ultimately improve the safety and accuracy of surgeries where tumor boundaries are visually indistinct.Chemistry and Chemical Biolog

Live-cell imaging of alkyne-tagged small biomolecules by stimulated Raman scattering

Sensitive and specific visualization of small biomolecules in living systems is highly challenging. We report stimulated Raman-scattering imaging of alkyne tags as a general strategy for studying a broad spectrum of small biomolecules in live cells and animals. We demonstrate this technique by tracking alkyne-bearing drugs in mouse tissues and visualizing de novo synthesis of DNA, RNA, proteins, phospholipids and triglycerides through metabolic incorporation of alkyne-tagged small precursors

Label-free, multi-scale imaging of ex-vivo mouse brain using spatial light interference microscopy

Brain connectivity spans over broad spatial scales, from nanometers to centimeters. In order to understand the brain at multi-scale, the neural network in wide-field has been visualized in detail by taking advantage of light microscopy. However, the process of staining or addition of fluorescent tags is commonly required, and the image contrast is insufficient for delineation of cytoarchitecture. To overcome this barrier, we use spatial light interference microscopy to investigate brain structure with high-resolution, sub-nanometer pathlength sensitivity without the use of exogenous contrast agents. Combining wide-field imaging and a mosaic algorithm developed in-house, we show the detailed architecture of cells and myelin, within coronal olfactory bulb and cortical sections, and from sagittal sections of the hippocampus and cerebellum. Our technique is well suited to identify laminar characteristics of fiber tract orientation within white matter, e.g. the corpus callosum. To further improve the macro-scale contrast of anatomical structures, and to better differentiate axons and dendrites from cell bodies, we mapped the tissue in terms of its scattering property. Based on our results, we anticipate that spatial light interference microscopy can potentially provide multiscale and multicontrast perspectives of gross and microscopic brain anatomy.ope

Supermultiplexed optical imaging and barcoding with engineered polyynes

Optical multiplexing has a large impact in photonics, the life sciences and biomedicine. However, current technology is limited by a 'multiplexing ceiling' from existing optical materials. Here we engineered a class of polyyne-based materials for optical supermultiplexing. We achieved 20 distinct Raman frequencies, as 'Carbon rainbow', through rational engineering of conjugation length, bond-selective isotope doping and end-capping substitution of polyynes. With further probe functionalization, we demonstrated ten-color organelle imaging in individual living cells with high specificity, sensitivity and photostability. Moreover, we realized optical data storage and identification by combinatorial barcoding, yielding to our knowledge the largest number of distinct spectral barcodes to date. Therefore, these polyynes hold great promise in live-cell imaging and sorting as well as in high-throughput diagnostics and screening