Large depth-of-field fluorescence microscopy based on deep learning supported by Fresnel incoherent correlation holography

Author(s):

Wu, Peng; Zhang, Dejie; Yuan, Jing; Zeng, Shaoqun; Gong, Hui; Luo, Qingming & Yang, Xiaoquan

Abstract:

“Fluorescence microscopy plays an irreplaceable role in biomedicine. However,

limited depth of field (DoF) of fluorescence microscopy is always an obstacle of image quality,

especially when the sample is with an uneven surface or distributed in different depths. In

this manuscript, we combine deep learning with Fresnel incoherent correlation holography to

describe a method to obtain significant large DoF fluorescence microscopy. Firstly, the hologram

is restored by the Auto-ASP method from out-of-focus to in-focus in double-spherical wave

Fresnel incoherent correlation holography. Then, we use a generative adversarial network to

eliminate the artifacts introduced by Auto-ASP and output the high-quality image as a result.

We use fluorescent beads, USAF target and mouse brain as samples to demonstrate the large

DoF of more than 400µm, which is 13 times better than that of traditional wide-field microscopy.

Moreover, our method is with a simple structure, which can be easily combined with many

existing fluorescence microscopic imaging technology”

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Publication: Optics Express
Issue/Year: Optics Express, Volume 30; Number 4; Pages 5177; 2022
DOI: 10.1364/oe.451409

Improving Multiphoton Microscopy by Combining Spherical Aberration Patterns and Variable Axicons

Author(s):

Bueno, J.M.; Hernández, G.; Skorsetz, M.; Artal, P.

Abstract:

“Multiphoton (MP) microscopy is a well-established method for the non-invasive imaging of biological tissues. However, its optical sectioning capabilities are reduced due to specimen-induced aberrations. Both the manipulation of spherical aberration (SA) and the use of axicons have been reported to be useful techniques to bypass this limitation. We propose the combination of SA patterns and variable axicons to further improve the quality of MP microscopy images. This approach provides enhanced images at different depth locations whose quality is better than those corresponding to the use of SA or axicons separately. Thus, the procedure proposed herein facilitates the visualization of details and increases the depth observable at high resolution.”

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Publication: Photonics
Issue/Year: Photonics, Volume 8; Number 12; Pages 573; 2021
DOI: 10.3390/photonics8120573

Complex-amplitude single-pixel imaging using coherent structured illumination

Author(s):

Hou, Hong-Yun; Zhao, Ya-Nan; Han, Jia-Cheng; Cui, Sheng-Wei; Cao, De-Zhong; Liu, Hong-Chao; Zhang, Su-Heng & Liang, Bao-Lai

Abstract:

“This research presents a coherent structured illumination single-pixel imaging scheme to image objects with complex amplitudes. By utilizing a phase-only spatial light modulator for phase modulation, we can efficiently generate the Hadamard basis structured light and the reference light that interfere with each other to form the coherent structured illumination. Using the 4-step phase-shifting, the spectrum of the object is acquired by detecting the zero-frequency component of the object light with a single-pixel photodetector. The desired complex-amplitude image can be further retrieved by applying an inverse Hadamard transform. The proposed scheme is experimentally demonstrated by imaging two etched glass objects, a dragonfly wing, and a resolution test chart. Benefiting from the phase modulation, this scheme has a high efficiency, a high imaging quality, a high spatial resolution, and a simple and stable configuration to obtain both the phase and amplitude information of the target object. The proposed scheme provides a promising complex-amplitude imaging modality with single-pixel detection. Thus it might find broad applications in optical metrology and biomedical science.”

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Publication: Optics Express
Issue/Year: Optics Express, Volume 29; Number 25; Pages 41827; 2021
DOI: 10.1364/oe.443258

Axial resolution enhancement for planar Airy beam light-sheet microscopy via the complementary beam subtraction method

Author(s):

Liu, Chao; Yu, Xianghua; Bai, Chen; Li, Xing; Zhou, Yuan; Yan, Shaohui; Min, Junwei; Dan, Dan; Li, Runze; Gu, Shuangyu & Yao, Baoli

Abstract:

“Airy beam light-sheet illumination can extend the field of view (FOV) of light-sheet fluorescence microscopy due to the unique propagation properties of non-diffraction and self-acceleration. However, the side lobes create undesirable out-of-focus background, leading to poor axial resolution and low image contrast. Here, we propose an Airy complementary beam subtraction (ACBS) method to improve the axial resolution while keeping the extended FOV. By scanning the optimized designed complementary beam that has two main lobes (TML), the generated complementary light-sheet has almost identical intensity distribution to that of the planar Airy light-sheet except for the central lobe. Subtraction of the two images acquired by double exposure respectively using the planar Airy light-sheet and the planar TML light-sheet can effectively suppress the influence of the out-of-focus background. The axial resolution improves from ∼4µm to 1.2 µm. The imaging performance was demonstrated by imaging specimens of aspergillus conidiophores and GFP labeled mouse brain section. The results show that the ACBS method enables the Airy beam light-sheet fluorescence microscopy to obtain better imaging quality.”

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Publication: Applied Optics
Issue/Year: Applied Optics, Volume 60; Number 32; Pages 10239; 2021
DOI: 10.1364/ao.441070

Towards non-blind optical tweezing by finding 3D refractive index changes through off-focus interferometric tracking

Author(s):

Landenberger, Benjamin; Yatish & Rohrbach, Alexander

Abstract:

“In modern 3D microscopy, holding and orienting arbitrary biological objects with optical forces instead of using coverslips and gel cylinders is still a vision. Although optical trapping forces are strong enough and related photodamage is acceptable, the precise (re-) orientation of large specimen with multiple optical traps is difficult, since they grab blindly at the object and often slip off. Here, we present an approach to localize and track regions with increased refractive index using several holographic optical traps with a single camera in an off-focus position. We estimate the 3D grabbing positions around several trapping foci in parallel through analysis of the beam deformations, which are continuously measured by defocused camera images of cellular structures inside cell clusters. Although non-blind optical trapping is still a vision, this is an important step towards fully computer-controlled orientation and feature-optimized laser scanning of sub-mm sized biological specimen for future 3D light microscopy.”

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Publication: Nature Communications
Issue/Year: Nature Communications, Volume 12; Number 1; 2021
DOI: 10.1038/s41467-021-27262-z

25D microscopy with polarization independent SLM for enhanced detection efficiency and aberration correction

Author(s):

Ren, Jinhan & Han, Kyu Young

Abstract:

“Fast, volumetric imaging by fluorescence microscopy is essential in studying bi-ological phenomena and cellular functions. Recently, single-shot 2.5D microscopy showedpromising results for high-throughput quantitative subcellular analysis via extended depth offield imaging without sequentialz-scanning; however, the detection efficiency was limited and itlacked depth-induced aberration correction. Here we report that a spatial light modulator (SLM)in a polarization insensitive configuration can significantly improve the detection efficiency of2.5D microscopy, while also compensating for aberrations at large imaging depths caused bythe refractive index mismatch between the sample and the immersion medium. We highlightthe improved efficiency via quantitative single-molecule RNA imaging of mammalian cellswith a 2-fold improvement in the fluorescence intensity compared to a conventional SLM-basedmicroscopy. We demonstrate the aberration correction capabilities and extended depth of field byimaging thick specimens with fewerz-scanning steps.”

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Publication: Optics Express
Issue/Year: Optics Express, Volume 29; Number 17; Pages 27530; 2021
DOI: 10.1364/oe.434260

Manipulating aqueous droplets by light-induced virtual electrodes

Author(s):

Zamboni, Riccardo; Imbrock, Jörg & Denz, Cornelia

Abstract:

“The precise spatio-temporal manipulation of droplets is fundamental for many lab-on-a-chip systems with applications in biology, healthcare and chemistry. Different approaches have been investigated, including thermal, chemical and electrical methodologies. Among this latter, electrophoresis (EP) and dielectrophoresis (DEP) play a key role, since they are highly compatible with microfluidic systems and provide sufficiently strong forces to control up to microliter volume aqueous droplets. However, EP and DEP techniques typically require the presence of metallic electrodes to create the desired electric fields, making these approaches less flexible and efficient than those exploiting pure optical techniques. Iron-doped lithium niobate (LiNbO3:Fe) allows for the generation of strong electric field modulation due to an inhomogenous illumination, thanks to its photovoltaic properties. These photoinduced fields interact as EP and DEP forces with microdroplets, while guaranteeing the flexibility provided by optical field-based modulation. Indeed, the combination with well-known techniques to control and modulate light fields can be exploited to generate virtual electrodes on the material, achieving reliable as well as flexible devices for water droplets control. In our approach, the photoinduced fields generated by the complex illumination of LiNbO3:Fe are exploited to control motion and trajectory of water droplets inside microfluidic channel. Moreover, the crystal is integrated in standard droplet microfluidic polymeric device, substituting the usual glass substrate and, thus without hindering the portability. This feature combined with the control of positions of aqueous droplets represents a key tool for several applications of customized lab-on-a-chip systems, highlighting the capabilities of LinbO3:Fe-based virtual electrodes.”

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Publication: SPIE Proceedings
Issue/Year: Proc. SPIE 11798, Optical Trapping and Optical Micromanipulation XVIII, 1179824, 2021
DOI: 10.1117/12.2594165

Optically Manipulated Microtools to Measure Adhesion of the Nanoparticle-Targeting Ligand Glutathione to Brain Endothelial Cells

Author(s):

Tamás Fekete, Mária Mészáros, Zsolt Szegletes, Gaszton Vizsnyiczai, László Zimányi, Mária A. Deli, Szilvia Veszelka*, and Lóránd Kelemen

Abstract:

“Targeting nanoparticles as drug delivery platforms is crucial to facilitate their cellular entry. Docking of nanoparticles by targeting ligands on cell membranes is the first step for the initiation of cellular uptake. As a model system, we studied brain microvascular endothelial cells, which form the anatomical basis of the blood–brain barrier, and the tripeptide glutathione, one of the most effective targeting ligands of nanoparticles to cross the blood–brain barrier. To investigate this initial docking step between glutathione and the membrane of living brain endothelial cells, we applied our recently developed innovative optical method. We present a microtool, with a task-specific geometry used as a probe, actuated by multifocus optical tweezers to characterize the adhesion probability and strength of glutathione-coated surfaces to the cell membrane of endothelial cells. The binding probability of the glutathione-coated surface and the adhesion force between the microtool and cell membrane was measured in a novel arrangement: cells were cultured on a vertical polymer wall and the mechanical forces were generated laterally and at the same time, perpendicularly to the plasma membrane. The adhesion force values were also determined with more conventional atomic force microscopy (AFM) measurements using functionalized colloidal probes. The optical trapping-based method was found to be suitable to measure very low adhesion forces (≤ 20 pN) without a high level of noise, which is characteristic for AFM measurements in this range. The holographic optical tweezers-directed functionalized microtools may help characterize the adhesion step of nanoparticles initiating transcytosis and select ligands to target nanoparticles.”

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Publication: ACS Applied Materials & Interfaces
Issue/Year: ACS Applied Materials & Interfaces, Volume 13; Number 33; Pages 39018–39029; 2021
DOI: 10.1021/acsami.1c08454

Super-resolution imaging by optical incoherent synthetic aperture with one channel at a time

Author(s):

Bulbul, Angika & Rosen, Joseph

Abstract:

“Imaging with an optical incoherent synthetic aperture (SA) means that the incoherent light from observed objects is processed over time from various points of view to obtain a resolution equivalent to single-shot imaging by the SA larger than the actual physical aperture. The operation of such systems has always been based on two-wave interference where the beams propagate through two separate channels. This limitation of two channels at a time is removed in the present study with the proposed SA where the two beams pass through the same single channel at any given time. The system is based on a newly developed self-interference technique named coded aperture correlation holography. At any given time, the recorded intensity is obtained from interference between two waves co-propagating through the same physical channel. One wave oriented in a particular polarization is modulated by a pseudorandom coded phase mask and the other one oriented orthogonally passes through an open subaperture. Both subapertures are multiplexed at the same physical window. The system is calibrated by a point spread hologram synthesized from the responses of a guide star. All the measurements are digitally processed to achieve a final image with a resolution higher than that obtained by the limited physical aperture. This unique configuration can offer alternatives for the current cumbersome systems composed of far apart optical channels in the large optical astronomical interferometers. Furthermore, the proposed concept paves the way to an SA system with a single less-expensive compact light collector in an incoherent optical regime that may be utilized for future ground-based or space telescopes.”

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Publication: Photonics Research
Issue/Year: Photonics Research, Volume 9; Number 7; Pages 1172; 2021
DOI: 10.1364/prj.422381

Single-shot digital multiplexed holography for the measurement of deep shapes

Author(s):

Kozacki, Tomasz; Mikuła-Zdańkowska, Marta; Martinez-Carranza, Juan & Idicula, Moncy Sajeev

Abstract:

“This work develops a single-shot holographic profilometer that enables shape characterization of discontinuous deep surfaces. This is achieved by combining hologram frequency multiplexing and an illumination technique of complex amplitude in multi-incidence angle profilometer. Object illumination is carried out from seven directions simultaneously, where the radial angular coordinates of illumination plane waves obey the geometric series. It is shown that: (i) the illumination pattern provides the required frequency separation of all object wavefronts in transverse frequency space, which is necessary for hologram demultiplexing, and (ii) numerical generation of longitudinal scanning function (LSF) is possible, which has large measurement range, high axial resolution, and small side lobes. Low side lobes of LSF and the developed multiplexed field dependent aberration compensation method are essential to minimize the negative influence of speckle noise of single-shot capture on the measurement result. The utility of the proposed method is demonstrated with experimental measurement of heights of two step-like objects.”

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Publication: Optics Express
Issue/Year: Optics Express, Volume 29; Number 14; Pages 21965; 2021
DOI: 10.1364/oe.428419
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