Stimulated Raman scattering (SRS) microscopy allows for high-speed label-free chemical imaging of biomedical systems. The imaging sensitivity of SRS microscopy is limited to ~10 mM for endogenous biomolecules. Electronic pre-resonant SRS allows detection of sub-micromolar chromophores. However, label-free SRS detection of single biomolecules having extremely small Raman cross-sections (~10−30 cm2sr−1) remains unreachable. Here, we demonstrate plasmon-enhanced stimulated Raman scattering (PESRS) microscopy with single-molecule detection sensitivity. Incorporating pico-Joule laser excitation, background subtraction, and a denoising algorithm, we obtain robust single-pixel SRS spectra exhibiting single-molecule events, verified by using two isotopologues of adenine and further confirmed by digital blinking and bleaching in the temporal domain. To demonstrate the capability of PESRS for biological applications, we utilize PESRS to map adenine released from bacteria due to starvation stress. PESRS microscopy holds the promise for ultrasensitive detection and rapid mapping of molecular events in chemical and biomedical systems.
Being able to image chemical bonds with high sensitivity and speed, stimulated Raman scattering (SRS) microscopy has made a major impact in biomedical optics. However, it is well known that the standard SRS microscopy suffers from various backgrounds, limiting the achievable contrast, quantification and sensitivity. While many frequency-modulation (FM) SRS schemes have been demonstrated to retrieve the sharp vibrational contrast, they often require customized laser systems and/or complicated laser pulse shaping or introduce additional noise, thereby hindering wide adoption. Herein we report a simple but robust strategy for FM-SRS microscopy based on a popular commercial laser system and regular optics. Harnessing self-phase modulation induced self-balanced spectral splitting of picosecond Stokes beam propagating in standard single-mode silica fibers, a high-performance FM-SRS system is constructed without introducing any additional signal noise. Our strategy enables adaptive spectral resolution for background-free SRS imaging of Raman modes with different linewidths. The generality of our method is demonstrated on a variety of Raman modes with effective suppressing of backgrounds including non-resonant cross phase modulation and electronic background from two-photon absorption or pump-probe process. As such, our method is promising to be adopted by the SRS microscopy community for background-free chemical imaging.
- Award ID(s):
- 1904684
- Publication Date:
- NSF-PAR ID:
- 10149198
- Journal Name:
- Optics Express
- Volume:
- 28
- Issue:
- 10
- Page Range or eLocation-ID:
- Article No. 15663
- ISSN:
- 1094-4087; OPEXFF
- Publisher:
- Optical Society of America
- Sponsoring Org:
- National Science Foundation
More Like this
-
Abstract -
One of the top priorities in observational astronomy is the direct imaging and characterization of extrasolar planets (exoplanets) and planetary systems. Direct images of rocky exoplanets are of particular interest in the search for life beyond the Earth, but they tend to be rather challenging targets since they are orders-of-magnitude dimmer than their host stars and are separated by small angular distances that are comparable to the classical
diffraction limit, even for the coming generation of 30 m class telescopes. Current and planned efforts for ground-based direct imaging of exoplanets combine high-order adaptive optics (AO) with a stellar coronagraph observing at wavelengths ranging from the visible to the mid-IR. The primary barrier to achieving high contrast with current direct imaging methods is quasi-static speckles, caused largely by non-common path aberrations (NCPAs) in the coronagraph optical train. Recent work has demonstrated that millisecond imaging, which effectively “freezes” the atmosphere’s turbulent phase screens, should allow the wavefront sensor (WFS) telemetry to be used as a probe of the optical system to measure NCPAs. Starting with a realistic model of a telescope with an AO system and a stellar coronagraph, this paper provides simulations of several closely related regression models that take advantagemore » -
The success of nonlinear optics relies largely on pulse-to-pulse consistency. In contrast, covariance-based techniques used in photoionization electron spectroscopy and mass spectrometry have shown that a wealth of information can be extracted from noise that is lost when averaging multiple measurements. Here, we apply covariance-based detection to nonlinear optical spectroscopy, and show that noise in a femtosecond laser is not necessarily a liability to be mitigated, but can act as a unique and powerful asset. As a proof of principle we apply this approach to the process of stimulated Raman scattering in α-quartz. Our results demonstrate how nonlinear processes in the sample can encode correlations between the spectral components of ultrashort pulses with uncorrelated stochastic fluctuations. This in turn provides richer information compared with the standard nonlinear optics techniques that are based on averages over many repetitions with well-behaved laser pulses. These proof-of-principle results suggest that covariance-based nonlinear spectroscopy will improve the applicability of fs nonlinear spectroscopy in wavelength ranges where stable, transform-limited pulses are not available, such as X-ray free-electron lasers which naturally have spectrally noisy pulses ideally suited for this approach.
-
A spatial heterodyne Raman spectrometer (SHRS), constructed using a modular optical cage and lens tube system, is described for use with a commercial silica and a custom single-crystal (SC) sapphire fiber Raman probe. The utility of these fiber-coupled SHRS chemical sensors is demonstrated using 532 nm laser excitation for acquiring Raman measurements of solid (sulfur) and liquid (cyclohexane) Raman standards as well as real-world, plastic-bonded explosives (PBX) comprising 1,3,5- triamino- 2,4,6- trinitrobenzene (TATB) and octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) energetic materials. The SHRS is a fixed grating-based dispersive interferometer equipped with an array detector. Each Raman spectrum was extracted from its corresponding fringe image (i.e., interferogram) using a Fourier transform method. Raman measurements were acquired with the SHRS Littrow wavelength set at the laser excitation wavelength over a spectral range of ∼1750 cm−1with a spectral resolution of ∼8 cm−1for sapphire and ∼10 cm−1for silica fiber probes. The large aperture of the SHRS allows much larger fiber diameters to be used without degrading spectral resolution as demonstrated with the larger sapphire collection fiber diameter (330 μm) compared to the silica fiber (100 μm). Unlike the dual silica fiber Raman probe, the dual sapphire fiber Raman probe did not include filtering at the fiber probe tip nearest the sample. Even so,more »
-
Abstract: 100 words Jurors are increasingly exposed to scientific information in the courtroom. To determine whether providing jurors with gist information would assist in their ability to make well-informed decisions, the present experiment utilized a Fuzzy Trace Theory-inspired intervention and tested it against traditional legal safeguards (i.e., judge instructions) by varying the scientific quality of the evidence. The results indicate that jurors who viewed high quality evidence rated the scientific evidence significantly higher than those who viewed low quality evidence, but were unable to moderate the credibility of the expert witness and apply damages appropriately resulting in poor calibration. Summary: <1000 words Jurors and juries are increasingly exposed to scientific information in the courtroom and it remains unclear when they will base their decisions on a reasonable understanding of the relevant scientific information. Without such knowledge, the ability of jurors and juries to make well-informed decisions may be at risk, increasing chances of unjust outcomes (e.g., false convictions in criminal cases). Therefore, there is a critical need to understand conditions that affect jurors’ and juries’ sensitivity to the qualities of scientific information and to identify safeguards that can assist with scientific calibration in the courtroom. The current project addresses thesemore »