A rigid coherent anti-Stokes Raman scattering endoscope with high resolution and a large field of view

Nonlinear optical endoscopy is an attractive technique for biomedical imaging since it promises to give access to high resolution imaging in vivo. Among the various techniques used for endoscopic contrast generation, coherent anti-Stokes Raman scattering (CARS) is especially interesting. CARS endoscopy allows molecule specific imaging of unlabeled samples. In this contribution, we describe the design, implementation, and experimental characterization of a rigid, compact CARS endoscope with a spatial resolution of 750 nm over a field of view of roughly 250 μm. Omission of the relay optics and use of a gradient index lens specifically designed for this application allow one to realize these specifications in an endoscopic unit which is 2.2 mm wide over a length of 187 mm, making clinical applications during surgical interventions possible. Multimodal use of the endoscope is demonstrated with images of samples with neurosurgical relevance.Nonlinear optical endoscopy is an attractive technique for biomedical imaging since it promises to give access to high resolution imaging in vivo. Among the various techniques used for endoscopic contrast generation, coherent anti-Stokes Raman scattering (CARS) is especially interesting. CARS endoscopy allows molecule specific imaging of unlabeled samples. In this contribution, we describe the design, implementation, and experimental characterization of a rigid, compact CARS endoscope with a spatial resolution of 750 nm over a field of view of roughly 250 μm. Omission of the relay optics and use of a gradient index lens specifically designed for this application allow one to realize these specifications in an endoscopic unit which is 2.2 mm wide over a length of 187 mm, making clinical applications during surgical interventions possible. Multimodal use of the endoscope is demonstrated with images of samples with neurosurgical relevance

ADPβS evokes microglia activation in the rabbit retina in vivo

To investigate whether stimulation of purinergic P2Y1 receptors modulates the activation of microglial and Müller glial cells in the rabbit retina in vivo, adenosine 5-O-(2-thiodiphosphate) (ADPβS; 2 mM in 100 μl saline), a non-hydrolyzable ADP analogue, was intravitreadly applied into control eyes or onto retinas that were experimentally detached from the pigment epithelium. Both retinal detachment and application of ADPßS onto control retinas induced phenotype alterations of the microglial cells (decrease of soma size, retraction of cell processes) and had no influence on microglial cell density. ADPßS application onto detached retinas accelerated the process retraction and resulted in a strongly decreased density of microglial cells. The effects of ADPßS on microglia density and phenotype in detached retinas were partially reversed by co-application of the selective inhibitor of P2Y1 receptors, MRS-2317 (3 mM in 100 μl saline). ADPßS apparently did not influence Müller cell gliosis, as determined by electrophysiological and calcium imaging records. It is concluded that rabbit retinal microglial cells express functional P2Y1 receptors in vivo, and that activation of these receptors stimulates phenotype alterations that are characteristical for microglia activation

Radiation enhancement and "temperature" in the collapse regime of gravitational scattering

We generalize the semiclassical treatment of graviton radiation to gravitational scattering at very large energies $\sqrt{s}\gg m_P$ and finite scattering angles $\Theta_s$, so as to approach the collapse regime of impact parameters $b \simeq b_c \sim R\equiv 2G\sqrt{s}$. Our basic tool is the extension of the recently proposed, unified form of radiation to the ACV reduced-action model and to its resummed-eikonal exchange. By superimposing that radiation all-over eikonal scattering, we are able to derive the corresponding (unitary) coherent-state operator. The resulting graviton spectrum, tuned on the gravitational radius $R$, fully agrees with previous calculations for small angles $\Theta_s\ll 1$ but, for sizeable angles $\Theta_s(b)\leq \Theta_c = O(1)$ acquires an exponential cutoff of the large $\omega R$ region, due to energy conservation, so as to emit a finite fraction of the total energy. In the approach-to-collapse regime of $b\to b_c^+$ we find a radiation enhancement due to large tidal forces, so that the whole energy is radiated off, with a large multiplicity $\langle N \rangle\sim Gs \gg 1$ and a well-defined frequency cutoff of order $R^{-1}$. The latter corresponds to the Hawking temperature for a black hole of mass notably smaller than $\sqrt{s}$.Comment: 5 pages, 2 figures, talk presented at the European Physical Society Conference on High Energy Physics, 5-12 July, Venice, Ital

Infrared Spectroscopic Studies of Cells and Tissues: Triple Helix Proteins as a Potential Biomarker for Tumors

In this work, the infrared (IR) spectra of living neural cells in suspension, native brain tissue, and native brain tumor tissue were investigated. Methods were developed to overcome the strong IR signal of liquid water so that the signal from the cellular biochemicals could be seen. Measurements could be performed during surgeries, within minutes after resection. Comparison between normal tissue, different cell lineages in suspension, and tumors allowed preliminary assignments of IR bands to be made. The most dramatic difference between tissues and cells was found to be in weaker IR absorbances usually assigned to the triple helix of collagens. Triple helix domains are common in larger structural proteins, and are typically found in the extracellular matrix (ECM) of tissues. An algorithm to correct offsets and calculate the band heights and positions of these bands was developed, so the variance between identical measurements could be assessed. The initial results indicate the triple helix signal is surprisingly consistent between different individuals, and is altered in tumor tissues. Taken together, these preliminary investigations indicate this triple helix signal may be a reliable biomarker for a tumor-like microenvironment. Thus, this signal has potential to aid in the intra-operational delineation of brain tumor borders. © 2013 Stelling et al

Imaging Arm Regeneration : Label-Free Multiphoton Microscopy to Dissect the Process in Octopus vulgaris

Cephalopod mollusks are endowed with an impressive range of features that have captured the attention of scientists from different fields, the imaginations of artists, and the interests of the public. The ability to spontaneously regrow lost or damaged structures quickly and functionally is among one of the most notable peculiarities that cephalopods possess. Microscopical imaging techniques represent useful tools for investigating the regenerative processes in several species, from invertebrates to mammals. However, these techniques have had limited use in cephalopods mainly due to the paucity of specific and commercially available markers. In addition, the commonly used immunohistochemical staining methods provide data that are specific to the antigens studied. New microscopical methods were recently applied to vertebrates to investigate regenerative events. Among them, multiphoton microscopy appears promising. For instance, it does not depend on species-related epitopes, taking advantage of the specific characteristics of tissues and allowing for its use in a species-independent way. Here, we illustrate the results obtained by applying this label-free imaging technique to the injured arm of Octopus vulgaris, a complex structure often subject to injury in the wild. This approach allowed for the characterization of the entire tissue arm architecture (muscular layers, nerve component, connective tissues, etc.) and elements usually hardly detectable (such as vessels, hemocytes, and chromatophores). More importantly, it also provided morpho-chemical information which helped decipher the regenerative phases after damage, from healing to complete arm regrowth, thereby appearing promising for regenerative studies in cephalopods and other non-model species.publishe

Combined Brillouin and Raman system for biomedical applications

A VIPA based Brillouin spectrometer consisting of commercially available components has been combined with a standard Raman spectrometer and a confocal microscope allowing simultaneous and co-localized measurements of biological samples and tissues

Combined Brillouin and Raman system for biomedical applications

A VIPA based Brillouin spectrometer consisting of commercially available components has been combined with a standard Raman spectrometer and a confocal microscope allowing simultaneous and co-localized measurements of biological samples and tissues

Learned end-to-end high-resolution lensless fiber imaging towards real-time cancer diagnosis

Recent advances in label-free histology promise a new era for real-time diagnosis in neurosurgery. Deep learning using autofluorescence is promising for tumor classification without histochemical staining process. The high image resolution and minimally invasive diagnostics with negligible tissue damage is of great importance. The state of the art is raster scanning endoscopes, but the distal lens optics limits the size. Lensless fiber bundle endoscopy offers both small diameters of a few 100 microns and the suitability as single-use probes, which is beneficial in sterilization. The problem is the inherent honeycomb artifacts of coherent fiber bundles (CFB). For the first time, we demonstrate an end-to-end lensless fiber imaging with exploiting the near-field. The framework includes resolution enhancement and classification networks that use single-shot CFB images to provide both high-resolution imaging and tumor diagnosis. The well-trained resolution enhancement network not only recovers high-resolution features beyond the physical limitations of CFB, but also helps improving tumor recognition rate. Especially for glioblastoma, the resolution enhancement network helps increasing the classification accuracy from 90.8 to 95.6%. The novel technique enables histological real-time imaging with lensless fiber endoscopy and is promising for a quick and minimally invasive intraoperative treatment and cancer diagnosis in neurosurgery