Diffuse optical spectroscopy and imaging to detect and quantify adipose tissue browning

Adipose (fat) tissue is a complex metabolic organ that is highly active and essential. In contrast to white adipose tissue (WAT), brown adipose tissue (BAT) is deemed metabolically beneficial because of its ability to burn calories through heat production. The conversion of WAT-resident adipocytes to “beige” or “brown-like” adipocytes has recently attracted attention. However, it typically takes a few days to analyze and confirm this browning of WAT through conventional molecular, biochemical, or histological methods. Moreover, accurate quantification of the overall browning process is not possible by any of these methods. In this context, we report the novel application of diffuse reflectance spectroscopy (DRS) and multispectral imaging (MSI) to detect and quantify the browning process in mice. We successfully demonstrated the time-dependent increase in browning of WAT, following its induction through β-adrenergic agonist injections. The results from these optical techniques were confirmed with those of standard molecular and biochemical assays, which measure gene and protein expression levels of UCP1 and PGC-1α, as well as with histological examinations. We envision that the reported optical methods can be developed into a fast, real time, cost effective and easy to implement imaging approach for quantification of the browning process in adipose tissue

Seeing Elastin: A Near-Infrared Zwitterionic Fluorescent Probe for In Vivo Elastin Imaging

Elastic fibers are present in a variety of tissues and are responsible for their resilience. Until now, no optical contrast agent in the near-infrared (NIR) wavelength range of 700-900 nm has been reported for the imaging of elastic fibers. Here, we report the discovery of a NIR zwitterionic elastin probe ElaNIR (elastin NIR) through fluorescent-image-based screening. The probe was successfully applied for in vitro, ex vivo, and in vivo imaging by various imaging modalities. Age-related elastin differences shown by in vivo fluorescent and photoacoustic imaging indicated that ElaNIR can be a potentially convenient tool for uncovering changes of elastin in live models.11Ysciescopu

Sensitive multiplex detection of serological liver cancer biomarkers using SERS-active photonic crystal fiber probe

Surface-enhanced Raman scattering (SERS) spectroscopy possesses the most promising advantage of multiplex detection for biosensing applications, which is achieved due to the narrow fingerprint' Raman spectra from the analyte molecules. We developed an ultrasensitive platform for the multiplex detection of cancer biomarkers by combining the SERS technique with a hollow-core photonic crystal fiber (HCPCF). Axially aligned air channels inside the HCPCF provide an excellent platform for optical sensing using SERS. In addition to the flexibility of optical fibers, HCPCF provides better light confinement and a larger interaction length for the guided light and the analyte, resulting in an improvement in sensitivity to detect low concentrations of bioanalytes in extremely low sample volumes. Herein, for the first time, we demonstrate the sensitive multiplex detection of biomarkers immobilized inside the HCPCF using antibody-conjugated SERS-active nanoparticles (SERS nanotags). As a proof-of-concept for targeted multiplex detection, initially we carried out the sensing of epidermal growth factor receptor (EGFR) biomarker in oral squamous carcinoma cell lysate using three different SERS nanotags. Subsequently, we also achieved simultaneous detection of hepatocellular carcinoma (HCC) biomarkers-alpha fetoprotein (AFP) and alpha-1-antitrypsin (A1AT) secreted in the supernatant from Hep3b cancer cell line. Using a SERS-HCPCF sensing platform, we could successfully demonstrate the multiplex detection in an extremely low sample volume of approximate to 20 nL. In future, this study may lead to sensitive biosensing platform for the low concentration detection of biomarkers in an extremely low sample volume of body fluids to achieve early diagnosis of multiple diseases. ((c) 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)1127sciescopu

Actively targeted in vivo multiplex detection of intrinsic cancer biomarkers using biocompatible sers nanotags

Surface-enhanced Raman scattering (SERS) technique is becoming highly popular for multiplex biosensing due to the \u27fingerprint\u27 Raman spectra from every molecule. As a proof-of-concept, we demonstrated the actively targeted multiplex in vitro and in vivo detection of three intrinsic cancer biomarkers - EGFR, CD44 and TGF beta RII in a breast cancer model using three multiplexing capable, biocompatible SERS nanoparticles/nanotags. Intra-tumorally injected antibody conjugated nanotags specifically targeting the three biomarkers exhibited maximum signal at 6 hours and no detectable signal at 72 hours. However, nanotags without antibodies showed no detectable signal after 6 hours. This difference could be due to the specific binding of the bioconjugated nanotags to the receptors on the cell surface. Thus, this study establishes SERS nanotags as an ultrasensitive nanoprobe for the multiplex detection of biomarkers and opens up its potential application in monitoring tumor progression and therapy and development into a theranostic probe

Optimized sandwiched surface plasmon resonance enhanced biosensor for multiplex biomarker detection

In this study, we performed finite element method (FEM) simulations to optimize the configuration of gold nanorods (GNR) enhanced surface plasmon resonance (SPR) sensor and discovered its application for multiplex antigens detection. Our work analyzed the near-field coupling between the sensing film and GNR. By systematically study the effect of gold film thickness, GNR-to-film distance and GNR dimensions on SPR, it was found that for GNR width smaller than 40nm, length change in GNR brought about significant SPR wavelength shift on the sensor, while the sensor is insensitive for GNR-to-film distance. As an application, we adopted GNRs of width 20 nm and aspect ratios from 2 to 4 and demonstrated the concept of conjugating gold film and GNRs with anti-Immunoglobulin G (anti-IgG) antibodies for multiplex detection of various IgG proteins with more than 100nm separation on their SPR wavelengths

Surface-enhanced raman scattering (sers)-based volatile organic compounds (vocs) detection using plasmonic bimetallic nanogap substrate

In this paper, we present surface-enhanced Raman scattering (SERS)-based volatile organic compounds (VOCs) detection with bimetallic nanogap structure substrate. Deep UV photolithography at the wavelength of 250 nm is used to pattern circular shape nanostructures. The nanogap between adjacent circular patterns is 30 +/- 5 nm. Silver (30 nm) and gold (15 nm) plasmonic active layers are deposited on the nanostructures subsequently. SERS measurements on different concentrations of acetone vapor ranged from 0.7, 1.5, 3.5, 10.3, 24.5 % and control have been performed with the substrate. The measurement results are found reproducible, and the detection limit is found to be 9.5 pg (acetone molecule). The detection sensitivity is 28.7 % higher than that of the recent reported leaning silicon nanopillar substrate. With further system miniaturization, the sensing technique can work as a portable SERS-based VOCs detection platform for point-of-care breath analysis, homeland security, chemical sensing and environmental monitoring

Machine Learning Assisted Real-Time Label-Free SERS Diagnoses of Malignant Pleural Effusion due to Lung Cancer

More than half of all pleural effusions are due to malignancy of which lung cancer is the main cause. Pleural effusions can complicate the course of pneumonia, pulmonary tuberculosis, or underlying systemic disease. We explore the application of label-free surface-enhanced Raman spectroscopy (SERS) as a point of care (POC) diagnostic tool to identify if pleural effusions are due to lung cancer or to other causes (controls). Lung cancer samples showed specific SERS spectral signatures such as the position and intensity of the Raman band in different wave number region using a novel silver coated silicon nanopillar (SCSNP) as a SERS substrate. We report a classification accuracy of 85% along with a sensitivity and specificity of 87% and 83%, respectively, for the detection of lung cancer over control pleural fluid samples with a receiver operating characteristics (ROC) area under curve value of 0.93 using a PLS-DA binary classifier to distinguish between lung cancer over control subjects. We have also evaluated discriminative wavenumber bands responsible for the distinction between the two classes with the help of a variable importance in projection (VIP) score. We found that our label-free SERS platform was able to distinguish lung cancer from pleural effusions due to other causes (controls) with higher diagnostic accuracy