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1. Photonic crystal enhanced fluorescence emission and blinking suppression for single quantum dot digital resolution biosensing

   https://doi.org/10.1038/s41467-022-32387-w  

   Xiong, Yanyu; Huang, Qinglan; Canady, Taylor D.; Barya, Priyash; Liu, Shengyan; Arogundade, Opeyemi H.; Race, Caitlin M.; Che, Congnyu; Wang, Xiaojing; Zhou, Lifeng; et al (December 2022, Nature Communications)

   Abstract While nanoscale quantum emitters are effective tags for measuring biomolecular interactions, their utilities for applications that demand single-unit observations are limited by the requirements for large numerical aperture (NA) objectives, fluorescence intermittency, and poor photon collection efficiency resulted from omnidirectional emission. Here, we report a nearly 3000-fold signal enhancement achieved through multiplicative effects of enhanced excitation, highly directional extraction, quantum efficiency improvement, and blinking suppression through a photonic crystal (PC) surface. The approach achieves single quantum dot (QD) sensitivity with high signal-to-noise ratio, even when using a low-NA lens and an inexpensive optical setup. The blinking suppression capability of the PC improves the QDs on-time from 15% to 85% ameliorating signal intermittency. We developed an assay for cancer-associated miRNA biomarkers with single-molecule resolution, single-base mutation selectivity, and 10-attomolar detection limit. Additionally, we observed differential surface motion trajectories of QDs when their surface attachment stringency is altered by changing a single base in a cancer-specific miRNA sequence. 

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2. Extended depth-of-field light-sheet microscopy improves imaging of large volumes at high numerical aperture

   https://doi.org/10.1063/5.0101426  

   Keomanee-Dizon, Kevin; Jones, Matt; Luu, Peter; Fraser, Scott_E; Truong, Thai_V (October 2022, Applied Physics Letters)

   Light-sheet microscopes must compromise among field of view, optical sectioning, resolution, and detection efficiency. High-numerical-aperture (NA) detection objective lenses provide higher resolution, but their narrow depth of field inefficiently captures the fluorescence signal generated throughout the thickness of the illumination light sheet when imaging large volumes. Here, we present ExD-SPIM (extended depth-of-field selective-plane illumination microscopy), an improved light-sheet microscopy strategy that solves this limitation by extending the depth of field (DOF) of high-NA detection objectives to match the thickness of the illumination light sheet. This extension of the DOF uses a phase mask to axially stretch the point-spread function of the objective lens while largely preserving lateral resolution. This matching of the detection DOF to the illumination-sheet thickness increases the total fluorescence collection, reduces the background, and improves the overall signal-to-noise ratio (SNR), as shown by numerical simulations, imaging of bead phantoms, and imaging living animals. In comparison to conventional light sheet imaging with low-NA detection that yields equivalent DOF, the results show that ExD-SPIM increases the SNR by more than threefold and dramatically reduces the rate of photobleaching. Compared to conventional high-NA detection, ExD-SPIM improves the signal sensitivity and volumetric coverage of whole-brain activity imaging, increasing the number of detected neurons by over a third. 

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3. Resolution enhancement of 2-photon microscopy using high-refractive index microspheres

   https://doi.org/10.1117/12.2290613  

   Tehrani, Kayvan F.; Darafsheh, Arash; Phang, Sendy; Mortensen, Luke J. (February 2018, Proc. SPIE 10498, Multiphoton Microscopy in the Biomedical Sciences XVIII)

   Intravital microscopy using multiphoton processes is the standard tool for deep tissue imaging inside of biological specimens. Usually, near-infrared and infrared light is used to excite the sample, which enables imaging several mean free path inside a scattering tissues. Using longer wavelengths, however, increases the width of the effective multiphoton Point Spread Function (PSF). Many features inside of cells and tissues are smaller than the diffraction limit, and therefore not possible to distinguish using a large PSF. Microscopy using high refractive index microspheres has shown promise to increase the numerical aperture of an imaging system and enhance the resolution. It has been shown that microspheres can image features ~λ/7 using single photon process fluorescence. In this work, we investigate resolution enhancement for Second Harmonic Generation (SHG) and 2-photon fluorescence microscopy. We used Barium Titanate glass microspheres with diameters ∼20–30 μm and refractive index ∼1.9–2.1. We show microsphere-assisted SHG imaging in bone collagen fibers. Since bone is a very dense tissue constructed of bundles of collagen fibers, it is nontrivial to image individual fibers. We placed microspheres on a dense area of the mouse cranial bone, and achieved imaging of individual fibers. We found that microsphere assisted SHG imaging resolves features of the bone fibers that are not readily visible in conventional SHG imaging. We extended this work to 2-photon microscopy of mitochondria in mouse soleus muscle, and with the help of microsphere resolving power, we were able to trace individual mitochondrion from their ensemble. 

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4. High-speed multiview imaging approaching 4pi steradians using conic section mirrors: theoretical and practical considerations

   https://doi.org/10.1364/JOSAA.440592  

   Zhou, Kevin_C; Dhalla, Al-Hafeez; McNabb, Ryan_P; Qian, Ruobing; Farsiu, Sina; Izatt, Joseph_A (November 2021, Journal of the Optical Society of America A)

   Illuminating or imaging samples from a broad angular range is essential in a wide variety of computational 3D imaging and resolution-enhancement techniques, such as optical projection tomography, optical diffraction tomography, synthetic aperture microscopy, Fourier ptychographic microscopy, structured illumination microscopy, photogrammetry, and optical coherence refraction tomography. The wider the angular coverage, the better the resolution enhancement or 3D-resolving capabilities. However, achieving such angular ranges is a practical challenge, especially when approaching $\pm <\#comment/>{90}^{\circ <\#comment/>}$or beyond. Often, researchers resort to expensive, proprietary high numerical aperture (NA) objectives or to rotating the sample or source-detector pair, which sacrifices temporal resolution or perturbs the sample. Here, we propose several new strategies for multiangle imaging approaching 4pi steradians using concave parabolic or ellipsoidal mirrors and fast, low rotational inertia scanners, such as galvanometers. We derive theoretically and empirically relations between a variety of system parameters (e.g.,  NA, wavelength, focal length, telecentricity) and achievable fields of view (FOVs) and importantly show that intrinsic tilt aberrations donotrestrict FOV for many multiview imaging applications, contrary to conventional wisdom. Finally, we present strategies for avoiding spherical aberrations at obliquely illuminated flat boundaries. Our simple designs allow for high-speed multiangle imaging for microscopic, mesoscopic, and macroscopic applications. 

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5. Broadband light extraction from near-surface NV centers using crystalline-silicon antennas

   Kim, Minjeong; Zahedian, Maryam; Wu, Wenxin; Fang, Chengyu; Yu, Zhaoning; Wambold, Raymond A; Yin, Shenwei; Czaplewski, David A; Choy, Jennifer T; Kats, Mikhail A (September 2024, arXivorg)

   We use crystalline silicon (Si) antennas to efficiently extract broadband single-photon fluorescence from shallow nitrogen-vacancy (NV) centers in diamond into free space. Our design features relatively easy-to-pattern high-index Si resonators on the diamond surface to boost photon extraction by overcoming total internal reflection and Fresnel reflection at the diamond-air interface, and providing modest Purcell enhancement, without etching or otherwise damaging the diamond surface. In simulations, ~20 times more single photons are collected from a single NV center compared to the case without the antenna; in experiments, we observe an enhancement of ~4 times, limited by spatial alignment between the NV and the antenna. Our approach can be readily applied to other color centers in diamond, and more generally to the extraction of light from quantum emitters in wide-bandgap materials. 

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