Comparing the fundamental imaging depth limit of two-photon, three-photon, and non-degenerate two-photon microscopy

Sadegh, Sanaz; Zilpelwar, Sharvari; Devor, Anna; Tian, Lei; Boas, David A.

doi:10.1364/ol.392724

Cited by 23 publications

(26 citation statements)

References 18 publications

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“…SBR total /SBR 2P shows the SBR improvement resulting from the 3PE contribution. Because the background for 3PE is negligible at these imaging depths (3,25), the background contribution is entirely from 2PE when a mixture of 2PE and 3PE exists. The calculated SBR upon 1340-nm excitation is 4.6×, 15×, and 82× larger than that at 1300, 1260, and 1220 nm (pure 2PE), respectively, because of the smaller contribution from 2PE at 1340 nm ( fig.…”

Section: Comparison Of Mouse Brain Imaging At Different Wavelengthsmentioning

confidence: 99%

Multicolor three-photon fluorescence imaging with single-wavelength excitation deep in mouse brain

2021

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Multiphoton fluorescence microscopy is a powerful technique for deep-tissue observation of living cells. In particular, three-photon microscopy is highly beneficial for deep-tissue imaging because of the long excitation wavelength and the high nonlinear confinement in living tissues. Because of the large spectral separation of fluorophores of different color, multicolor three-photon imaging typically requires multiple excitation wavelengths. Here, we report a new three-photon excitation scheme: excitation to a higher-energy electronic excited state instead of the conventional excitation to the lowest-energy excited state, enabling multicolor three-photon fluorescence imaging with deep-tissue penetration in the living mouse brain using single-wavelength excitation. We further demonstrate that our excitation method results in ≥10-fold signal enhancement for some of the common red fluorescent molecules. The multicolor imaging capability and the possibility of enhanced three-photon excitation cross section will open new opportunities for life science applications of three-photon microscopy.

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Section: Comparison Of Mouse Brain Imaging At Different Wavelengthsmentioning

confidence: 99%

Multicolor three-photon fluorescence imaging with single-wavelength excitation deep in mouse brain

2021

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“…Cheng et al 49 . The key idea of the BPM is to treat the scattering material as a series of planes that orthogonal to the initial light propagation direction.…”

Section: Modeling For the Two-photon Laser Beam Propagationmentioning

confidence: 99%

“…The key idea of the BPM is to treat the scattering material as a series of planes that orthogonal to the initial light propagation direction. At each plane, we simulate the scattering by multiplying the local wavefront by a random phase term 49,50 . Here we adopt a 2D-cylindrical BPM model with laser propagated along the z axis to predict the wavefront in the water and brain (Supplementary Fig.…”

Section: Modeling For the Two-photon Laser Beam Propagationmentioning

confidence: 99%

Probing neuropeptide volume transmission in vivo by a novel all-optical approach

Xiong

Lacin

Ouyang

et al. 2021

Preprint

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Neuropeptides are essential signaling molecules in the nervous system involved in modulating neural circuits and behavior. Although hypothesized to signal via volume transmission through G-protein coupled receptors (GPCR), remarkably little is known about their extrasynaptic diffusion. Here, we developed an all-optical approach to probe neuropeptide volume transmission in mouse neocortex. To control neuropeptide release, we engineered photosensitive nanovesicles with somatostatin-14 (SST) that is released with near-infrared light stimulation. To detect SST, we created a new cell-based neurotransmitter fluorescent engineered reporter (CNiFER) using the SST2 GPCR. Under two-photon imaging, we determined the time to activate SST2R at defined distances as well as the maximal distance and loss rate for SST volume transmission in neocortex. Importantly, we determined that SST transmission is significantly faster in neocortex with a chemically degraded extracellular matrix, a diseased condition indicated in neuroinflammation and Parkinson′s disease. These new neurotechnologies can reveal important biological signaling processes previously not possible, and provide new opportunities to investigate volume transmission in the brain.

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“…1B). Modern FLI and PLIM platforms cover an optical resolution ranging from sub-micrometer to several millimeters, a spectral range from 180 to 1700 nm and a time-domain resolution from sub-picoseconds to milliseconds, with measurement speeds that range from real-time for one-photon FLI and PLIM (Raspe et al, 2016;Trinh et al, 2019;Cheng et al, 2020) to several minutes per frame for two-photon PLIM (Rytelewski et al, 2019). By allowing quantitative, multiparameter and often non-invasive and nondestructive three-dimensional (3D) analyses of live cells and tissues to be performed, FLI and PLIM provide a broad support to the ongoing '3D molecular-imaging revolution'.…”

Section: Introductionmentioning

confidence: 99%

“…2). These approaches are mainly associated with microscopy techniques that are used on transparent and/or thin samples, allowing for good light penetration depth (Cheng et al, 2020). However, when considering intact biological tissues, scattering becomes a prevalent phenomenon, precluding the ability to confine the detected photon to a localized 2D plane.…”

Section: Introductionmentioning

confidence: 99%

Luminescence lifetime imaging of three-dimensional biological objects

Dmitriev

Intes

Barroso

2021

Journal of Cell Science

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A major focus of current biological studies is to fill the knowledge gaps between cell, tissue and organism scales. To this end, a wide array of contemporary optical analytical tools enable multiparameter quantitative imaging of live and fixed cells, three-dimensional (3D) systems, tissues, organs and organisms in the context of their complex spatiotemporal biological and molecular features. In particular, the modalities of luminescence lifetime imaging, comprising fluorescence lifetime imaging (FLI) and phosphorescence lifetime imaging microscopy (PLIM), in synergy with Förster resonance energy transfer (FRET) assays, provide a wealth of information. On the application side, the luminescence lifetime of endogenous molecules inside cells and tissues, overexpressed fluorescent protein fusion biosensor constructs or probes delivered externally provide molecular insights at multiple scales into protein–protein interaction networks, cellular metabolism, dynamics of molecular oxygen and hypoxia, physiologically important ions, and other physical and physiological parameters. Luminescence lifetime imaging offers a unique window into the physiological and structural environment of cells and tissues, enabling a new level of functional and molecular analysis in addition to providing 3D spatially resolved and longitudinal measurements that can range from microscopic to macroscopic scale. We provide an overview of luminescence lifetime imaging and summarize key biological applications from cells and tissues to organisms.

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Comparing the fundamental imaging depth limit of two-photon, three-photon, and non-degenerate two-photon microscopy

Cited by 23 publications

References 18 publications

Multicolor three-photon fluorescence imaging with single-wavelength excitation deep in mouse brain

Multicolor three-photon fluorescence imaging with single-wavelength excitation deep in mouse brain

Probing neuropeptide volume transmission in vivo by a novel all-optical approach

Luminescence lifetime imaging of three-dimensional biological objects

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