Nanoscopy enables breaking down the light diffraction limit and reveals the nanostructures of objects being studied using light. In 2014, three scientists pioneered the development of nanoscopy and won the Nobel Prize in Chemistry. This recognized the achievement of the past twenty years in the field of nanoscopy. However, fluorescent probes used in the field of nanoscopy are still numbered. Here, we review the currently available four categories of probes and existing methods to improve the performance of probes.
A hyperspectral image data cube acquired from HEK‐293 cells labeled with cytoplasmic and nuclear stains: Calcein Green and NucBlu. The top view (XY plane) displays three spectrally unmixed channels for cellular autofluorescence (red), Calcein Green (green), and NucBlue (blue). The Z axis shows spectral information, from low to high wavelength. The article by Leavesley and colleagues describes an approach for calculating the sensitivity of spectral imaging assays for detecting a fluorescence signature within a mix of other signatures or autofluorescence. Further details can be found in the article by Silas J. Leavesley et al. ( e201600227 ).
Germanium vs Silicon: All‐dielectric nanoparticles provides the heat resistance for proteins under light‐induced heating. Further details can be found in the article by Andrei A. Krasilin et al. ( e201700322 )
An integrated 4‐modality endoscopy system combining white light imaging, autofluorescence imaging, diffuse reflectance spectroscopy and Raman spectroscopy technologies was developed for in vivo endoscopic nasopharyngeal cancer detection. Both high diagnostic sensitivity (98.6%) and high specificity (95.1%) for differentiating cancer from normal tissue sites were achieved using this system combined with multivariate diagnostic algorithm, demonstrating great potential for improving real‐time, in vivo diagnosis of cancer at endoscopy. Further details can be found in the article by Duo Lin et al. ( e201700251 )
Optical coherence tomography through an implanted dorsal imaging window allows for prolonged in vivo structural and functional assessment of the mouse oviduct (Fallopian tube), including threedimensional structural imaging, quantitative measurements of the smooth muscle contraction, and mapping of cilia beat frequency. This method brings new opportunities for live studies and longitudinal analyses of mouse reproductive events in the native context. Further details can be found in the article by Shang Wang et al. ( e201700316 ).
A plasmon waveguide resonance (PWR) sensor is proposed for studying the interaction between gold nanoparticles and proteins. The ability of the PWR sensor to operate in both TM and TE Polarizations, i.e. its polarization diversity, facilitates the simultaneous spectroscopy of the nanoparticles surface reactions using both polarizations. The response of each polarization to streptavidin‐biotin binding at the surface of gold nanoparticles is investigated in real time. Finally, using the principles of multimode spectroscopy, the nanoparticle's surface reactions are decoupled from the bulk solution refractive index variations.
Schematic diagram of the NP‐modified PWR sensor 相似文献
In this paper the utilization of smartphone as a detection platform for colorimetric quantification of biological macromolecules has been demonstrated. Using V‐channel of HSV color space, the quantification of BSA protein, catalase enzyme and carbohydrate (using D‐glucose) have been successfully investigated. A custom designed android application has been developed for estimating the total concentration of biological macromolecules. The results have been compared with that of a standard spectrophotometer which is generally used for colorimetric quantification in laboratory settings by measuring its absorbance at a specific wavelength. The results obtained with the designed sensor is found to be similar when compared with the spectrophotometer data. The designed sensor is low cost, robust and we envision that it could promote diverse fields of bio‐analytical investigations.
Schematic illustration of the smartphone sensing mechanism for colorimetric analysis of biomolecular samples. 相似文献
In vivo multiphoton imaging was used to map changes in hepatobiliary metabolism in liver fibrosis (left column) and hepatocellular carcinoma (right column). The top row shows the maps of kinetic rate constant of the uptake and esterase processing while the bottom row shows that of bile canalicular excretion of xenobiotics. Further details can be found in the article by Chih‐Ju Lin, Sheng‐Lin Lee, Wei‐Hsiang Wang, et al. ( e201700338 ).
Raman images were used to study the effect of the contaminant chlorpyriphos‐oxon on zebrafish eye samples. Multivariate Curve Resolution‐Alternating Least Squares (MCR‐ALS) was used to obtain the distribution maps and spectral signatures of biological components present in the images analyzed. The use of MCRALS spectral signatures as starting information for Partial Least Squares‐Discriminant Analysis allowed statistical assessment of the effect of the contaminant at a specific tissue level. Further details can be found in the article by Víctor Olmos et al. ( e201700089 ).
In this study, we introduce two key improvements that overcome limitations of existing polygon scanning microscopes while maintaining high spatial and temporal imaging resolution over large field of view (FOV). First, we proposed a simple and straightforward means to control the scanning angle of the polygon mirror to carry out photomanipulation without resorting to high speed optical modulators. Second, we devised a flexible data sampling method directly leading to higher image contrast by over 2‐fold and digital images with 100 megapixels (10 240 × 10 240) per frame at 0.25 Hz. This generates sub‐diffraction limited pixels (60 nm per pixels over the FOV of 512 μm) which increases the degrees of freedom to extract signals computationally. The unique combined optical and digital control recorded fine fluorescence recovery after localized photobleaching (r ~10 μm) within fluorescent giant unilamellar vesicles and micro‐vascular dynamics after laser‐induced injury during thrombus formation in vivo. These new improvements expand the quantitative biological‐imaging capacity of any polygon scanning microscope system.
Bladder cancer is among the most common cancers in the UK and conventional detection techniques suffer from low sensitivity, low specificity, or both. Recent attempts to address the disparity have led to progress in the field of autofluorescence as a means to diagnose the disease with high efficiency, however there is still a lot not known about autofluorescence profiles in the disease. The multi‐functional diagnostic system “LAKK‐M” was used to assess autofluorescence profiles of healthy and cancerous bladder tissue to identify novel biomarkers of the disease. Statistically significant differences were observed in the optical redox ratio (a measure of tissue metabolic activity), the amplitude of endogenous porphyrins and the NADH/porphyrin ratio between tissue types. These findings could advance understanding of bladder cancer and aid in the development of new techniques for detection and surveillance.
We disclose a theranostic device for performing image‐guided riboflavin/UV‐A corneal cross‐linking. The device determines treatment efficacy by real time monitoring of riboflavin concentration in the corneal stroma. The study shows efficacy of the device in eye bank human donor tissues. Further details can be found in the article by Giuseppe Lombardo et al. ( e201800028 )
Sensitive Escherichia coli detection based on a T4 bacteriophageimmobilized multimode microfiber is proposed and demonstrated in this article. Different modes are excited and guided in the microfiber as evanescent field that can interact with surrounding E. coli directly. The change of E. coli concentration and corresponding binding of E. coli on microfiber surface will lead to the shift of optical spectrum, which can be exploited for the application of biosensing. Further details can be found in the article by Yanpeng Li, Hui Ma, Lin Gan, et al. ( e201800012 ).
A label‐free interferometric transducer showing a theoretical detection limit for homogeneous sensing of 5 × 10–8 RIU, being equivalent to a protein mass coverage resolution of 2.8 fg mm–2, is used to develop a high sensitive biosensor for protein detection. The extreme sensitivity of this transducer combined with a selective bioreceptor layer enables the direct evaluation of the human growth hormone (hGH) in undiluted urine matrix in the 10 pg mL–1 range.
Congenital cardiovascular defects are the leading cause of birth defect related death. It has been hypothesized that fluid mechanical forces of embryonic blood flow affect cardiovascular development and play a role in congenital malformations. Studies in small animal embryos can improve our understanding of congenital malformations and can lead to better treatment. We present a feasibility study in which high‐resolution optical coherence tomography (OCT) and computational fluid dynamics (CFD) are combined to provide quantitative analysis of the embryonic flow mechanics and the associated anatomy in a small animal model.
Gold nanoparticles serve as imaging contrast agents useful for two‐photon nonlinear microscopy of biological cells and tissues. In this study, 100‐nm‐sized gold particles with a multitude of nanopores embedded inside have been physically synthesized and investigated for the plasmonic enhancement in two‐photon luminescence. Exhibiting remarkable potential for two‐photon imaging, the porous gold nanoparticles boost near‐infrared light absorption substantially and allow emission signals 20 times brighter than gold nanorods being currently used as typical imaging agents. Further details can be found in the article by Joo H. Park et al. ( e201700174 )
We present a hybrid dual‐wavelength optoacoustic and ultrasound bio‐microscope capable of rapid transcranial visualization of morphology and oxygenation status of large‐scale cerebral vascular networks. Imaging of entire cortical vasculature in mice is achieved with single capillary resolution and complemented by simultaneously acquired pulse‐echo ultrasound microscopy scans of the mouse skull. The new approach holds potential to facilitate studies into neurological and vascular abnormalities of the brain. Further details can be found in the article by Johannes Rebling, Héctor Estrada, Sven Gottschalk, et al. ( e201800057 ).
Eu3+integrated photoluminescence intensity ratio (PLIR) approach for optical detection of lactates in blood serum, plasma and confocal imaging of brain tissues has very high potential for exploitation of this technique in both in vitro monitoring and in vivo bioimaging applications for the detection of biomarkers in various diseases states. This image is diagrammatic representation of fact that the overall PLIR is higher with more lactates conjugated with Eu3+ ions. Further details can be found in the article by Tarun Kakkar et al. ( e201700199 ).
Tissue autofluorescence provides fluorescence lifetime contrast between acellular tissue and that containing newly seeded cells. Fiber‐based fluorescence lifetime imaging (FLIm) can be used for tracking recellularization of engineered vascular grafts and potential matrix remodeling at large scale, without compromising sample integrity. FLIm cellular contrast was verified in a subset of samples seeded with eGFP‐labelled cells. Results suggests fiberbased FLIm is a suitable tool for monitoring recellularization of engineered tissue nondestructively. Further details can be found in the article by Alba Alfonso‐Garcia, Jeny Shklover, Benjamin E. Sherlock, et al. ( e201700391 ).
下载免费PDF全文