Multicolor two-photon microscopy for living mice using broadband fiber-continuum selective excitation and linear unmixing

2021 ◽  
Author(s):  
Xinyuan Huang ◽  
Zhongyun Chen ◽  
Ling Fu
Keyword(s):  
Selective Excitation ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
Linear Unmixing

scholarly journals Robust blind spectral unmixing for fluorescence microscopy using unsupervised learning

2019 ◽  
Author(s):  
Tristan D. McRae ◽  
David Oleksyn ◽  
Jim Miller ◽  
Yu-Rong Gao
Keyword(s):  
Fluorescence Microscopy ◽  
Emission Spectra ◽  
Principal Component ◽  
Spectral Unmixing ◽  
Spectral Signatures ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
False Positive Detection ◽  
Linear Unmixing ◽  
Microscopy Images

AbstractDue to the overlapping emission spectra of fluorophores, fluorescence microscopy images often have bleed-through problems, leading to a false positive detection. This problem is almost unavoidable when the samples are labeled with three or more fluorophores, and the situation is complicated even further when imaged under a multiphoton microscope. Several methods have been developed and commonly used by biologists for fluorescence microscopy spectral unmixing, such as linear unmixing, non-negative matrix factorization, deconvolution, and principal component analysis. However, they either require pre-knowledge of emission spectra or restrict the number of fluorophores to be the same as detection channels, which highly limits the real-world applications of those spectral unmixing methods. In this paper, we developed a robust and flexible spectral unmixing method: Learning Unsupervised Means of Spectra (LUMoS), which uses an unsupervised machine learning clustering method to learn individual fluorophores’ spectral signatures from mixed images, and blindly separate channels without restrictions on the number of fluorophores that can be imaged. This method highly expands the hardware capability of two-photon microscopy to simultaneously image more fluorophores than is possible with instrumentation alone. Experimental and simulated results demonstrated the robustness of LUMoS in multi-channel separations of two-photon microscopy images. We also extended the application of this method to background/autofluorescence removal and colocalization analysis. Lastly, we integrated this tool into ImageJ to offer an easy to use spectral unmixing tool for fluorescence imaging. LUMoS allows us to gain a higher spectral resolution and obtain a cleaner image without the need to upgrade the imaging hardware capabilities.

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

scholarly journals Adaptive Optical Two-Photon Microscopy for Surface-Profiled Living Biological Specimens

ACS Omega ◽  
2020 ◽  
Author(s):  
Kazushi Yamaguchi ◽  
Kohei Otomo ◽  
Yuichi Kozawa ◽  
Motosuke Tsutsumi ◽  
Tomoko Inose ◽  
...  
Keyword(s):  
Two Photon Microscopy ◽  
Two Photon

scholarly journals Publisher Correction: Miniature two-photon microscopy for enlarged field-of-view, multi-plane and long-term brain imaging

2021 ◽  
Vol 18 (2) ◽  
pp. 220-220
Author(s):  
Weijian Zong ◽  
Runlong Wu ◽  
Shiyuan Chen ◽  
Junjie Wu ◽  
Hanbin Wang ◽  
...  
Keyword(s):  
Brain Imaging ◽  
Field Of View ◽  
Two Photon Microscopy ◽  
Two Photon

scholarly journals Publisher Correction: Ultrahigh-speed point scanning two-photon microscopy using high dynamic range silicon photomultipliers

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Vincent D. Ching-Roa ◽  
Eben M. Olson ◽  
Sherrif F. Ibrahim ◽  
Richard Torres ◽  
Michael G. Giacomelli
Keyword(s):  
Dynamic Range ◽  
High Dynamic Range ◽  
Silicon Photomultipliers ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
High Dynamic

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

scholarly journals Two-photon microscopy with simultaneous standard and extended depth of field using a tunable acoustic gradient-index lens

2009 ◽  
Vol 34 (11) ◽  
pp. 1684 ◽  
Author(s):  
Nicolas Olivier ◽  
Alexandre Mermillod-Blondin ◽  
Craig B. Arnold ◽  
Emmanuel Beaurepaire
Keyword(s):  
Depth Of Field ◽  
Gradient Index ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
Gradient Index Lens ◽  
Extended Depth Of Field

scholarly journals Evaluation of Nanoparticle Penetration in the Tumor Spheroid Using Two-Photon Microscopy

Biomedicines ◽  
2020 ◽  
Vol 9 (1) ◽  
pp. 10
Author(s):  
Feby Wijaya Pratiwi ◽  
Chien-Chung Peng ◽  
Si-Han Wu ◽  
Chiung Wen Kuo ◽  
Chung-Yuan Mou ◽  
...  
Keyword(s):  
Physicochemical Properties ◽  
Solid Tumors ◽  
Physicochemical Parameters ◽  
Mesoporous Silica Nanoparticles ◽  
Model System ◽  
Tumor Spheroid ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
Tumor Region ◽  
Penetration Behavior

Mesoporous silica nanoparticles (MSNs) have emerged as a prominent nanomedicine platform, especially for tumor-related nanocarrier systems. However, there is increasing concern about the ability of nanoparticles (NPs) to penetrate solid tumors, resulting in compromised antitumor efficacy. Because the physicochemical properties of NPs play a significant role in their penetration and accumulation in solid tumors, it is essential to systematically study their relationship in a model system. Here, we report a multihierarchical assessment of the accumulation and penetration of fluorescence-labeled MSNs with nine different physicochemical properties in tumor spheroids using two-photon microscopy. Our results indicated that individual physicochemical parameters separately could not define the MSNs’ ability to accumulate in a deeper tumor region; their features are entangled. We observed that the MSNs’ stability determined their success in reaching the hypoxia region. Moreover, the change in the MSNs’ penetration behavior postprotein crowning was associated with both the original properties of NPs and proteins on their surfaces.

Optimisation of fluorescent DNA labels for two-photon microscopy

2010 ◽  
Author(s):  
G. Metgé ◽  
C. Fiorini-Debuisschert ◽  
F. Charra ◽  
G. Bordeau ◽  
E. Faurel ◽  
...  
Keyword(s):  
Two Photon Microscopy ◽  
Two Photon
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