Nanoscale local modification of PMMA refractive index by tip-enhanced femtosecond pulsed laser irradiation

Investigation techniques based on tip-enhanced optical effects, capable to yield spatial resolutions down to nanometers level, have enabled a wide palette of important discoveries over the past twenty years. Recently, their underlying optical setups are beginning to emerge as useful tools to modify and manipulate matter with nanoscale spatial resolution. We try to contribute to these efforts by reporting a method that we found viable to modify the surface refractive index of polymethyl methacrylate (PMMA), an acrylic polymer material. The changes in the refractive index are accomplished by focusing a femtosecond pulsed near-infrared laser beam on the apex of a metalized nano-sized tip, traditionally used in scanning probe microscopy (SPM) applications. The adopted illumination strategy yields circular-shaped modifications of the refractive index occurring at the surface of the PMMA sample, exhibiting a lateral size <200 nm, under 790 nm illumination, representing a four-fold increase in precision compared to the current state-of-the-art. The light intensity enhancement effects taking place at the tip apex makes possible achieving refractive index changes at low laser pulse energies (<0.5 nJ), which represents two orders of magnitude advantage over the current state-of-the art. The presented nanoimprinting method is very flexible, as it can be used with different power levels and can potentially be operated with other materials. Besides enabling modifications of the refractive index with high lateral resolution, this method can pave the way towards other important applications such the fabrication of photonic crystal lattices or surface waveguides

Experimenting Liver Fibrosis Diagnostic by Two Photon Excitation Microscopy and Bag-of-Features Image Classification

The accurate staging of liver fibrosis is of paramount importance to determine the state of disease progression, therapy responses, and to optimize disease treatment strategies. Non-linear optical microscopy techniques such as two-photon excitation fluorescence (TPEF) and second harmonic generation (SHG) can image the endogenous signals of tissue structures and can be used for fibrosis assessment on non-stained tissue samples. While image analysis of collagen in SHG images was consistently addressed until now, cellular and tissue information included in TPEF images, such as inflammatory and hepatic cell damage, equally important as collagen deposition imaged by SHG, remain poorly exploited to date. We address this situation by experimenting liver fibrosis quantification and scoring using a combined approach based on TPEF liver surface imaging on a Thioacetamide-induced rat model and a gradient based Bag-of-Features (BoF) image classification strategy. We report the assessed performance results and discuss the influence of specific BoF parameters to the performance of the fibrosis scoring framework.Romania. Executive Agency for Higher Education, Research, Development and Innovation Funding (research grant PN-II-PT-PCCA-2011-3.2-1162)Rectors' Conference of the Swiss Universities (SCIEX NMS-CH research fellowship nr. 12.135)Singapore. Agency for Science, Technology and Research (R-185-000-182-592)Singapore. Biomedical Research CouncilInstitute of Bioengineering and Nanotechnology (Singapore)Singapore-MIT Alliance (Computational and Systems Biology Flagship Project funding (C-382-641-001-091))Singapore-MIT Alliance for Research and Technology (SMART BioSyM and Mechanobiology Institute of Singapore (R-714-001-003-271)

Mapping electron beam injected trapped charge with scattering scanning near-field optical microscopy

Scattering scanning near-field optical microscopy (s-SNOM) has been demonstrated as a valuable tool for mapping the optical and optoelectronic properties of materials with nanoscale resolution. Here we report experimental evidence that trapped electric charges injected by an electron beam at the surface of dielectric samples affect the sample-dipole interaction, which has direct impact on the s-SNOM image content. Nanoscale mapping of the surface trapped charge holds significant potential for the precise tailoring of the electrostatic properties of dielectric and semiconductive samples, such as hydroxyapatite, which has particular importance with respect to biomedical applications. The methodology developed here is highly relevant to semiconductor device fabrication as well

Roadmap on Label-Free Super-resolution Imaging

Label-free super-resolution (LFSR) imaging relies on light-scattering processes in nanoscale objects without a need for fluorescent (FL) staining required in super-resolved FL microscopy. The objectives of this Roadmap are to present a comprehensive vision of the developments, the state-of-the-art in this field, and to discuss the resolution boundaries and hurdles that need to be overcome to break the classical diffraction limit of the label-free imaging. The scope of this Roadmap spans from the advanced interference detection techniques, where the diffraction-limited lateral resolution is combined with unsurpassed axial and temporal resolution, to techniques with true lateral super-resolution capability that are based on understanding resolution as an information science problem, on using novel structured illumination, near-field scanning, and nonlinear optics approaches, and on designing superlenses based on nanoplasmonics, metamaterials, transformation optics, and microsphere-assisted approaches. To this end, this Roadmap brings under the same umbrella researchers from the physics and biomedical optics communities in which such studies have often been developing separately. The ultimate intent of this paper is to create a vision for the current and future developments of LFSR imaging based on its physical mechanisms and to create a great opening for the series of articles in this field.Peer reviewe

Effect of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker initiation on organ support-free days in patients hospitalized with COVID-19

IMPORTANCE Overactivation of the renin-angiotensin system (RAS) may contribute to poor clinical outcomes in patients with COVID-19. Objective To determine whether angiotensin-converting enzyme (ACE) inhibitor or angiotensin receptor blocker (ARB) initiation improves outcomes in patients hospitalized for COVID-19. DESIGN, SETTING, AND PARTICIPANTS In an ongoing, adaptive platform randomized clinical trial, 721 critically ill and 58 non–critically ill hospitalized adults were randomized to receive an RAS inhibitor or control between March 16, 2021, and February 25, 2022, at 69 sites in 7 countries (final follow-up on June 1, 2022). INTERVENTIONS Patients were randomized to receive open-label initiation of an ACE inhibitor (n = 257), ARB (n = 248), ARB in combination with DMX-200 (a chemokine receptor-2 inhibitor; n = 10), or no RAS inhibitor (control; n = 264) for up to 10 days. MAIN OUTCOMES AND MEASURES The primary outcome was organ support–free days, a composite of hospital survival and days alive without cardiovascular or respiratory organ support through 21 days. The primary analysis was a bayesian cumulative logistic model. Odds ratios (ORs) greater than 1 represent improved outcomes. RESULTS On February 25, 2022, enrollment was discontinued due to safety concerns. Among 679 critically ill patients with available primary outcome data, the median age was 56 years and 239 participants (35.2%) were women. Median (IQR) organ support–free days among critically ill patients was 10 (–1 to 16) in the ACE inhibitor group (n = 231), 8 (–1 to 17) in the ARB group (n = 217), and 12 (0 to 17) in the control group (n = 231) (median adjusted odds ratios of 0.77 [95% bayesian credible interval, 0.58-1.06] for improvement for ACE inhibitor and 0.76 [95% credible interval, 0.56-1.05] for ARB compared with control). The posterior probabilities that ACE inhibitors and ARBs worsened organ support–free days compared with control were 94.9% and 95.4%, respectively. Hospital survival occurred in 166 of 231 critically ill participants (71.9%) in the ACE inhibitor group, 152 of 217 (70.0%) in the ARB group, and 182 of 231 (78.8%) in the control group (posterior probabilities that ACE inhibitor and ARB worsened hospital survival compared with control were 95.3% and 98.1%, respectively). CONCLUSIONS AND RELEVANCE In this trial, among critically ill adults with COVID-19, initiation of an ACE inhibitor or ARB did not improve, and likely worsened, clinical outcomes. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT0273570

Generic arrays of surface-positioned and shallow-buried gold multi-shapes as reference samples to benchmark near-field microscopes. Part 1: Applications in s-SNOM depth imaging

A wide palette of nanoscale imaging techniques operating in the near-field regime has been reported to date, enabling an important number of scientific breakthroughs. While the tuning and benchmarking of near-field microscopes represent a very important step for optimizing the outputs of the imaging sessions, no generally acknowledged standards exist yet in terms of calibration of near-field microscopes, which would play an important role in fully exploiting the potential of these instruments. With this work, we aim to contribute to filling in this gap, by introducing a prototypical sample, that holds potential for becoming a benchmark with respect to comparing the performances of diverse near-field measurement techniques, including traditional, aperture based, scanning near field microscopy (SNOM), or apertureless variants, such as scattering-type scanning nearfield optical microscopy (s-SNOM). The proposed samples have been thoroughly simulated, and an easy fabrication procedure is presented and demonstrated. In this latter context, Au-SiO2 samples sharing different configurations, in terms of geometry, number and depth of contrast yielding layers, enabling both surface and sub-surface nanoscopy measurements, were designed and fabricated. We argue that the proposed prototypical samples can be highly useful for benchmarking the outputs of various near-field microscopy techniques, as they facilitate a broad range of tests, relevant for comparing the performances and accuracy of many diverse investigation methods. We also introduce a methodology for numerically simulating the samples and their near-field after illuminating them with light of different wavelengths, as well as their simple process flow. This methodology can considerably augment their future use as a prototypical sample for the evaluation and calibration of current and next generation near-field nanoscopy techniques. Experimental evidence on the usefulness of these samples as s-SNOM testing and benchmarking tools is provided in the context of differentiation of surface and sub-surface structures, and influence of tip-sample distance on attainable amplitude and phase signals. We consider these efforts to represent an important, required step, in advancing the near-field imaging field, with important potential to augment the outputs of current near-field imaging systems, and to facilitate the development and benchmarking of next generation of near-field instrumentation

Editorial: Advances in label free tissue imaging with laser scanning microscopy techniques

Advances in Label Free Tissue Imaging with Laser Scanning Microscopy Technique