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Register a new userDeep penetration fluorescence imaging through dense yeast cells suspensions using Airy beams
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We propose a new method to image fluorescent objects through turbid media base on Airy beam scanning. This is achieved by using the non-diffractive nature of Airy beams, namely their ability to maintain their shape while penetrating through a highly scattering medium. We show, that our technique can image fluorescent objects immersed in turbid media with higher resolution and signal to noise than confocal imaging. As proof-of-principle, we demonstrate imaging of 1$mu$m sized fluorescent beads through a dense suspension of yeast cells with an attenuation coefficient of 51cm$^{-1}$ at a depth of 90$mu$m. Finally, we demonstrate that our technique can also provide the depth of the imaged object without any additional sectioning.
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Subwavelength imaging by microsphere lenses is a promising label-free super-resolution imaging technique. There is a growing interest to use live cells to replace the widely used non-biological microsphere lenses. In this work, we demonstrate the use of yeast cells for such imaging purpose. Using fiber-based optical trapping technique, we successfully trapped a chain of yeast cells and bring them to the vicinity of imaging objects. These yeast cells work as near-field magnifying lenses and simultaneously pick up the sub-diffraction information of the nanoscale objects under each cell and project them into the far-field. Blu-ray disc of 100 nm feature can be clearly resolved in a parallel manner by each cell, thus effectively increasing the imaging field of view and imaging efficiency. Our work will contribute to the further development of more advanced bio-superlens imaging system
High-resolution optical microscopy suffers from a low contrast in scattering media where a multiply scattered wave obscures a ballistic wave used for image formation. To extend the imaging depth, various gating operations - confocal, coherence, and polarization gating - have been devised to filter out the multiply scattered wave. However, these gating methods are imperfect as they all act on the detection plane located outside a scattering medium. Here, we present a new gating scheme, called space gating, that rejects the multiply scattered wave directly at the object plane inside a scattering medium. Specifically, we introduced a 30 $mu$m-wide acoustic focus to the object plane and reconstructed a coherent image only with the ballistic wave modulated by acousto-optic interaction. This method allows us to reject the multiply scattered wave that the existing gating methods cannot filter out and improves the ratio of the ballistic wave to the multiply scattered wave by more than 100 times for a scattering medium more than 20 times thicker than its scattering mean free path. Using the coherent imaging technique based on space gating, we demonstrate the unprecedented imaging capability - phase imaging of optically transparent biological cells fully embedded within a scattering medium - with a spatial resolution of 1.5 $mu$m.
Beam self-cleaning (BSC) in graded-index (GRIN) multimode fibres (MMFs) has been recently reported by different research groups. Driven by the interplay between Kerr effect and beam self-imaging, BSC counteracts random mode coupling, and forces laser beams to recover a quasi-single mode profile at the output of GRIN fibres. Here we show that the associated self-induced spatiotemporal reshaping allows for improving the performances of nonlinear fluorescence microscopy and endoscopy using multimode optical fibres. We experimentally demonstrate that the beam brightness increase, induced by self-cleaning, enables two and three-photon imaging of biological samples with high spatial resolution. Temporal pulse shortening accompanying spatial beam clean-up enhances the output peak power, hence the efficiency of nonlinear imaging. We also show that spatiotemporal supercontinuum generation is well-suited for large-band nonlinear fluorescence imaging in visible and infrared domains. We substantiated our findings by multiphoton fluorescence imaging in both microscopy and endoscopy configurations.
Over the last dozen of years, the area of accelerating waves has made considerable advances not only in terms of fundamentals and experimental demonstrations but also in connection to a wide range of applications. Starting from the prototypical Airy beam that was proposed and observed in 2007, new families of accelerating waves have been identified in the paraxial and nonparaxial domains in space and/or time, with different methods developed to control at will their trajectory, amplitude, and beam width. Accelerating optical waves exhibit a number of highly desirable attributes. They move along a curved or accelerating trajectory while being resilient to perturbations (self-healing), and, are diffraction-free. It is because of these particular features that accelerating waves have been utilized in a variety of applications in the areas of filamentation, beam focusing, particle manipulation, biomedical imaging, plasmons, and material processing among others.
Super-resolution fluorescence microscopy is an important tool in biomedical research for its ability to discern features smaller than the diffraction limit. However, due to its difficult implementation and high cost, the universal application of super-resolution microscopy is not feasible. In this paper, we propose and demonstrate a new kind of super-resolution fluorescence microscopy that can be easily implemented and requires neither additional hardware nor complex post-processing. The microscopy is based on the principle of stepwise optical saturation (SOS), where $M$ steps of raw fluorescence images are linearly combined to generate an image with a $sqrt{M}$-fold increase in resolution compared with conventional diffraction-limited images. For example, linearly combining (scaling and subtracting) two images obtained at regular powers extends resolution by a factor of $1.4$ beyond the diffraction limit. The resolution improvement in SOS microscopy is theoretically infinite but practically is limited by the signal-to-noise ratio. We perform simulations and experimentally demonstrate super-resolution microscopy with both one-photon (confocal) and multiphoton excitation fluorescence. We show that with the multiphoton modality, the SOS microscopy can provide super-resolution imaging deep in scattering samples.
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