scholarly journals Poor man's two photon imaging: Scanning Laser Upconversion Microscopy

2017 ◽  
Author(s):  
Cecilia Sorbello ◽  
Roberto Etchenique
Keyword(s):  
Femtosecond Lasers ◽  
Laser Diodes ◽  
Scanning Laser ◽  
Deep Penetration ◽  
Multiphoton Absorption ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
Instantaneous Power ◽  
Optical Images ◽  
Photon Imaging

ABSTRACTTwo photon microscopy is one of the most powerful techniques for optical images. Its deep penetration, low scattering and mainly its sectioning power allows exquisite control on all three axes of 3D samples, both for imaging and actuation. Unfortunately, this technique involves very expensive femtosecond lasers. Upconverting Nanoparticles display multiphoton absorption at low instantaneous power, which can be achieved by means of cheap laser diodes. Scanning Laser Upconversion Microscopy offers the deep penetration and low scattering of NIR excitation together with the sectioning power of multiphoton techniques for under US$ 1000.

scholarly journals Multidither coherent optical adaptive technique for deep tissue two-photon microscopy

2019 ◽  
Vol 12 (04) ◽  
pp. 1942003 ◽  
Author(s):  
Biwei Zhang ◽  
Wei Gong ◽  
Chenxue Wu ◽  
Lejia Hu ◽  
Xinpei Zhu ◽  
...  
Keyword(s):  
Adaptive Optics ◽  
Diffraction Theory ◽  
Fluorescence Signal ◽  
Two Photon Microscopy ◽  
Scalar Diffraction ◽  
Two Photon ◽  
Scalar Diffraction Theory ◽  
Adaptive Technique ◽  
Photon Imaging ◽  
Two Photon Imaging

Two-photon microscopy normally suffers from the scattering of the tissue in biological imaging. Multidither coherent optical adaptive technique (COAT) can correct the scattered wavefront in parallel. However, the determination of the corrective phases may not be completely accurate using conventional method, which undermines the performance of this technique. In this paper, we theoretically demonstrate a method that can obtain more accurate corrective phases by determining the phase values from the square root of the fluorescence signal. A numerical simulation model is established to study the performance of adaptive optics in two-photon microscopy by combining scalar diffraction theory with vector diffraction theory. The results show that the distortion of the wavefront can be corrected more thoroughly with our method in two-photon imaging. In our simulation, with the scattering from a 450-[Formula: see text]m-thick mouse brain tissue, excitation focal spots with higher peak-to-background ratio (PBR) and images with higher contrast can be obtained. Hence, further enhancement of the multidither COAT correction performance in two-photon imaging can be expected.

scholarly journals Volumetric two-photon imaging in live cells and embryos via axially gradient excitation

2019 ◽  
Author(s):  
Yufeng Gao ◽  
Xianyuan Xia ◽  
Jia Yu ◽  
Tingai Chen ◽  
Zhili Xu ◽  
...  
Keyword(s):  
Live Cells ◽  
Deep Penetration ◽  
Layer By Layer ◽  
Photon Beams ◽  
Volumetric Imaging ◽  
Localization Accuracy ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
Subcellular Resolution ◽  
Imaging Speed

AbstractTwo-photon microscopy(TPM) that features subcellular resolution, intrinsic optical sectioning ability, and deep penetration in sample is a powerful tool of bioimaging. However, the process of layer-by-layer scanning to form a 3D image inherently limits the volumetric imaging speed and significantly increases the phototoxicity. Here we develop a gradient TPM technique that enables rapid volumetric imaging by only acquiring two 2D images. By sequentially exciting the specimen with two axially elongated two-photon beams with complementary gradient intensities, the axial positions of fluorophores can be decoded from the intensity ratio of the paired images. We achieve an axial localization accuracy of 0.728 ± 0.657 μm, which is sufficient for rapid 3D subcellular imaging. We demonstrate the flexibility of the gradient TPM on a variety of sparsely labelled samples, including bead phantoms, mouse brain tissues, live macrophages and live nematode embryos. The results show that, compared with conventional TPM, the 3D imaging speed increases 6 folds while the photobleaching and photodamage are extremely reduced.

scholarly journals jYCaMP: An optimized calcium indicator for two-photon imaging at fiber laser wavelengths

2019 ◽  
Author(s):  
Manuel Alexander Mohr ◽  
Daniel Bushey ◽  
Abhi Aggarwal ◽  
Jonathan S. Marvin ◽  
Emiliano Jimenez Marquez ◽  
...  
Keyword(s):  
Fiber Laser ◽  
Calcium Imaging ◽  
Femtosecond Lasers ◽  
State Of The Art ◽  
Two Photon ◽  
Photon Excitation ◽  
Calcium Indicator ◽  
Calcium Indicators ◽  
Photon Imaging ◽  
Two Photon Excitation

AbstractState-of-the-art GFP-based calcium indicators do not undergo efficient two-photon excitation at wavelengths above 1000 nm, for which inexpensive and powerful industrial femtosecond lasers are available. Here we report jYCaMP1, a yellow variant of jGCaMP7 that outperforms its parent in mice and flies at excitation wavelengths above 1000 nm and enables improved two-color calcium imaging with RFP-based indicators.

scholarly journals Intravital imaging of adriamycin-induced renal pathology using two-photon microscopy and optical coherence tomography

2018 ◽  
Vol 11 (05) ◽  
pp. 1850030 ◽  
Author(s):  
Hengchang Guo ◽  
Hsing-Wen Wang ◽  
Qinggong Tang ◽  
Erik Anderson ◽  
Reuben Falola ◽  
...  
Keyword(s):  
Optical Coherence Tomography ◽  
Real Time ◽  
Flow Dynamics ◽  
Renal Pathology ◽  
Glomerular Sclerosis ◽  
Deep Penetration ◽  
Optical Coherence ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
Tubular Flow

Adriamycin (doxorubicin), a common cancer chemotherapeutic drug, can be used to induce a model of chronic progressive glomerular disease in rodents. In our studies, we evaluated renal changes in a rat model after Adriamycin injection using two-photon microscopy (TPM), optical coherence tomography (OCT) and Doppler OCT (DOCT). Taking advantage of deep penetration and fast scanning speed for three-dimensional (3D) label-free imaging, OCT/DOCT system was able to reveal glomerular and tubular pathology noninvasively and in real time. By imaging renal pathology following the infusion of fluorophore-labeled dextrans of different molecular weights, TPM can provide direct views of glomerular and tubular flow dynamics with the onset and progression of renal disease. Specifically, glomerular permeability and filtration, proximal and distal tubular flow dynamics can be revealed. 6–8 weeks after injection of Adriamycin, TPM and OCT/DOCT imaging revealed glomerular sclerosis, compromised flow across the glomerular wall, tubular atrophy, tubular dilation, and variable intra-tubular flow dynamics. Our results indicate that TPM and OCT/DOCT provide real-time imaging of renal pathology in vivo that has not been previously available using conventional microscopic procedures.

An easily synthesized AIE luminogen for lipid droplet-specific super-resolution imaging and two-photon imaging

2021 ◽  
Author(s):  
Yanzi Xu ◽  
Haoke Zhang ◽  
Ning Zhang ◽  
Ruohan Xu ◽  
Zhi Wang ◽  
...  
Keyword(s):  
Penetration Depth ◽  
Lipid Droplet ◽  
Super Resolution ◽  
Deep Penetration ◽  
Two Photon ◽  
Resolution Imaging ◽  
Photon Imaging ◽  
Two Photon Imaging ◽  
Photo Stability ◽  
In Cells

A synergetic imaging platform was established to achieve LDs-specific imaging by STED nanoscopy and TPF microscopy. Benefiting from its high PLQYs, outstanding photo-stability, and high LDs specificity, a superior resolution in cells and a deep penetration depth in tissues were achieved.

In vivo imaging of liver injury and regeneration by functional two-photon microscopy

2016 ◽  
Vol 54 (12) ◽  
pp. 1343-1404
Author(s):  
A Ghallab ◽  
R Reif ◽  
R Hassan ◽  
AS Seddek ◽  
JG Hengstler
Keyword(s):  
Liver Injury ◽  
In Vivo Imaging ◽  
Two Photon Microscopy ◽  
Two Photon
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