scholarly journals Nanoscopic resolution within a single imaging frame

2021 ◽  
Author(s):  
Esley Torres ◽  
Raúl Pinto ◽  
Alejandro Linares ◽  
Damián Martínez ◽  
Víctor Abonza ◽  
...  
Keyword(s):  
Spatial Resolution ◽  
Live Cell Imaging ◽  
Laser Scanning ◽  
Cell Imaging ◽  
Mean Shift ◽  
Super Resolution ◽  
Theoretical Limit ◽  
The Mean ◽  
Scanning Systems ◽  
Imaging Conditions

Mean-Shift Super Resolution (MSSR) is a principle based on the Mean Shift theory that improves the spatial resolution in fluorescence images beyond the diffraction limit. MSSR works on low- and high-density fluorophore images, is not limited by the architecture of the detector (EM-CCD, sCMOS, or photomultiplier-based laser scanning systems) and is applicable to single images as well as temporal series. The theoretical limit of spatial resolution, based on optimized real-world imaging conditions and analysis of temporal image series, has been measured to be 40 nm. Furthermore, MSSR has denoising capabilities that outperform other analytical super resolution image approaches. Altogether, MSSR is a powerful, flexible, and generic tool for multidimensional and live cell imaging applications.

scholarly journals Nanoscopic resolution within a single imaging frame

2021 ◽  
Author(s):  
Esley García ◽  
Raúl Cámara ◽  
Alejandro Linares ◽  
Damián Martínez ◽  
Víctor Abonza ◽  
...  
Keyword(s):  
Spatial Resolution ◽  
Live Cell Imaging ◽  
Laser Scanning ◽  
Cell Imaging ◽  
Mean Shift ◽  
Super Resolution ◽  
Theoretical Limit ◽  
The Mean ◽  
Scanning Systems ◽  
Imaging Conditions

Abstract Mean-Shift Super Resolution (MSSR) is a principle based on the Mean Shift theory that improves the spatial resolution in fluorescence images beyond the diffraction limit. MSSR works on low- and high-density fluorophore images, is not limited by the architecture of the detector (EM-CCD, sCMOS, or photomultiplier-based laser scanning systems) and is applicable to single images as well as temporal series. The theoretical limit of spatial resolution, based on optimized real-world imaging conditions and analysis of temporal image series, has been measured to be 40 nm. Furthermore, MSSR has denoising capabilities that outperform other analytical super resolution image approaches. Altogether, MSSR is a powerful, flexible, and generic tool for multidimensional and live cell imaging applications.

scholarly journals Challenges in 3D Live Cell Imaging

Photonics ◽  
2021 ◽  
Vol 8 (7) ◽  
pp. 275
Author(s):  
Herbert Schneckenburger ◽  
Verena Richter
Keyword(s):  
Live Cell Imaging ◽  
Laser Scanning ◽  
Cell Imaging ◽  
Live Cell ◽  
Laser Scanning Microscopy ◽  
Radiative Energy ◽  
Wide Field ◽  
Optical Sectioning ◽  
Continuous Increase ◽  
Radiative Energy Transfer

A short overview on 3D live cell imaging is given. Relevant samples are described and various problems and challenges—including 3D imaging by optical sectioning, light scattering and phototoxicity—are addressed. Furthermore, enhanced methods of wide-field or laser scanning microscopy together with some relevant examples and applications are summarized. In the future one may profit from a continuous increase in microscopic resolution, but also from molecular sensing techniques in the nanometer range using e.g., non-radiative energy transfer (FRET).

scholarly journals Dynamic Organization of Chromatin Domains Revealed by Super-Resolution Live-Cell Imaging

2017 ◽  
Vol 67 (2) ◽  
pp. 282-293.e7 ◽  
Author(s):  
Tadasu Nozaki ◽  
Ryosuke Imai ◽  
Mai Tanbo ◽  
Ryosuke Nagashima ◽  
Sachiko Tamura ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Super Resolution ◽  
Live Cell ◽  
Chromatin Domains ◽  
Dynamic Organization

scholarly journals LRRK2 mediates tubulation and vesicle sorting from membrane damaged lysosomes

2020 ◽  
Author(s):  
Luis Bonet-Ponce ◽  
Alexandra Beilina ◽  
Chad D. Williamson ◽  
Eric Lindberg ◽  
Jillian H. Kluss ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Super Resolution ◽  
Adaptor Protein ◽  
Dependent Manner ◽  
Leucine Rich Repeat ◽  
Biological Functions ◽  
New Process ◽  
Sporadic Parkinson’S Disease

ABSTRACTMutations in the leucine rich repeat kinase 2 (LRRK2) gene are a cause of familial and sporadic Parkinson’s disease (PD). Nonetheless, the biological functions of LRRK2 remain incompletely understood. Here, we observed that LRRK2 is recruited to lysosomes that have a ruptured membrane. Using unbiased proteomics, we observed that LRRK2 is able to recruit the motor adaptor protein JIP4 to permeabilized lysosomes in a kinase-dependent manner through the phosphorylation of RAB35 and RAB10. Super-resolution live cell imaging microscopy and FIB-SEM revealed that once at the lysosomal membrane, JIP4 promotes the formation of LAMP1-negative lysosomal tubules that release membranous content from ruptured lysosomes. Released vesicular structures are able to interact with other lysosomes. Thus, we described a new process that uses lysosomal tubulation to release vesicular structures from permeabilized lysosomes. LRRK2 orchestrates this process that we name LYTL (LYsosomal Tubulation/sorting driven by LRRK2) that, given the central role of the lysosome in PD, is likely to be disease relevant.

scholarly journals C. elegans neurons have functional dendritic spines

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Andrea Cuentas-Condori ◽  
Ben Mulcahy ◽  
Siwei He ◽  
Sierra Palumbos ◽  
Mei Zhen ◽  
...  
Keyword(s):  
Motor Neurons ◽  
Dendritic Spines ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Super Resolution ◽  
Live Cell ◽  
Model Organisms ◽  
Spine Density ◽  
C Elegans ◽  
Spine Function

Dendritic spines are specialized postsynaptic structures that transduce presynaptic signals, are regulated by neural activity and correlated with learning and memory. Most studies of spine function have focused on the mammalian nervous system. However, spine-like protrusions have been reported in C. elegans (Philbrook et al., 2018), suggesting that the experimental advantages of smaller model organisms could be exploited to study the biology of dendritic spines. Here, we used super-resolution microscopy, electron microscopy, live-cell imaging and genetics to show that C. elegans motor neurons have functional dendritic spines that: (1) are structurally defined by a dynamic actin cytoskeleton; (2) appose presynaptic dense projections; (3) localize ER and ribosomes; (4) display calcium transients triggered by presynaptic activity and propagated by internal Ca++ stores; (5) respond to activity-dependent signals that regulate spine density. These studies provide a solid foundation for a new experimental paradigm that exploits the power of C. elegans genetics and live-cell imaging for fundamental studies of dendritic spine morphogenesis and function.

scholarly journals Malic Acid Carbon Dots: From Super-resolution Live-Cell Imaging to Highly Efficient Separation

ACS Nano ◽  
2018 ◽  
Vol 12 (6) ◽  
pp. 5741-5752 ◽  
Author(s):  
Bo Zhi ◽  
Yi Cui ◽  
Shengyang Wang ◽  
Benjamin P. Frank ◽  
Denise N. Williams ◽  
...  
Keyword(s):  
Malic Acid ◽  
Carbon Dots ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Super Resolution ◽  
Live Cell ◽  
Efficient Separation ◽  
Highly Efficient

scholarly journals Customizable live-cell imaging chambers for multimodal and multiplex fluorescence microscopy

2020 ◽  
Vol 98 (5) ◽  
pp. 612-623
Author(s):  
Adam Tepperman ◽  
David Jiao Zheng ◽  
Maria Abou Taka ◽  
Angela Vrieze ◽  
Austin Le Lam ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Super Resolution ◽  
Live Cell ◽  
Imaging Modalities ◽  
Experimental Conditions ◽  
Microscopy Imaging ◽  
Glass Coverslip ◽  
Multiple Imaging ◽  
Microscopy Techniques

Using multiple imaging modalities while performing independent experiments in parallel can greatly enhance the throughput of microscopy-based research, but requires the provision of appropriate experimental conditions in a format that meets the optical requirements of the microscope. Although customized imaging chambers can meet these challenges, the difficulty of manufacturing custom chambers and the relatively high cost and design inflexibility of commercial chambers has limited the adoption of this approach. Herein, we demonstrate the use of 3D printing to produce inexpensive, customized, live-cell imaging chambers that are compatible with a range of imaging modalities, including super-resolution microscopy. In this approach, biocompatible plastics are used to print imaging chambers designed to meet the specific needs of an experiment, followed by adhesion of the printed chamber to a glass coverslip, producing a chamber that is impermeant to liquids and that supports the growth and imaging of cells over multiple days. This approach can also be used to produce moulds for casting microfluidic devices made of polydimethylsiloxane. The utility of these chambers is demonstrated using designs for multiplex microscopy, imaging under shear, chemotaxis, and general cellular imaging. Together, this approach represents an inexpensive yet highly customizable approach for producing imaging chambers that are compatible with modern microscopy techniques.

A millisecond structured illumination microscope for super-resolution live cell imaging

2020 ◽  
Vol 13 (4) ◽  
pp. 045002
Author(s):  
Tomu Suzuki ◽  
Shinji Kajimoto ◽  
Narufumi Kitamura ◽  
Mayumi Takano-Kasuya ◽  
Naoko Furusawa ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Super Resolution ◽  
Live Cell ◽  
Structured Illumination
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