scholarly journals Advanced Methods in Fluorescence Microscopy

2013 ◽  
Vol 36 (1-2) ◽  
pp. 5-17 ◽  
Author(s):  
Luke Fritzky ◽  
David Lagunoff
Keyword(s):  
Fluorescence Microscopy ◽  
Laser Scanning ◽  
Fluorescent Proteins ◽  
Second Harmonic ◽  
Super Resolution ◽  
Good Deal ◽  
Light Sheet ◽  
Confocal Laser ◽  
Will Power ◽  
Super Resolution Microscopy

It requires a good deal of will power to resist hyperbole in considering the advances that have been achieved in fluorescence microscopy in the last 25 years. Our effort has been to survey the modalities of microscopic fluorescence imaging available to cell biologists and perhaps useful for diagnostic pathologists. The gamut extends from established confocal laser scanning through multiphoton and TIRF to the emerging technologies of super-resolution microscopy that breech the Abbé limit of resolution. Also considered are the recent innovations in structured and light sheet illumination, the use of FRET and molecular beacons that exploit specific characteristics of designer fluorescent proteins, fluorescence speckles, and second harmonic generation for native anisometric structures like collagen, microtubules and sarcomeres.

scholarly journals Meeting experiments at the diffraction barrier: an in-silico widefield fluorescence microscopy

2021 ◽  
Author(s):  
Subhamoy Mahajan ◽  
Tian Tang
Keyword(s):  
Fluorescence Microscopy ◽  
In Silico ◽  
Super Resolution ◽  
Light Sheet ◽  
Atomic Resolution ◽  
Live Cells ◽  
Two Photon ◽  
Color Combinations ◽  
Super Resolution Microscopy ◽  
Diffraction Barrier

AbstractFluorescence microscopy allows the visualization of live cells and their components, but even with advances in super- resolution microscopy, atomic resolution remains unattainable. On the other hand, molecular simulations (MS) can easily access atomic resolution, but comparison with experimental microscopy images has not been possible. In this work, a novel in-silico widefield fluorescence microscopy is proposed, which reduces the resolution of MS to generate images comparable to experiments. This technique will allow cross-validation and compound the knowledge gained from experiments and MS. We demonstrate that in-silico images can be produced with different optical axis, object focal planes, exposure time, color combinations, resolution, brightness and amount of out-of-focus fluorescence. This allows the generation of images that resemble those obtained from widefield, confocal, light-sheet, two-photon and super-resolution microscopy. This technique not only can be used as a standalone visualization tool for MS, but also lays the foundation for other in-silico microscopy methods.

Structural Evidence of Photoisomerization Pathways in Fluorescent Proteins

2019 ◽  
Author(s):  
Jeffrey Chang ◽  
Matthew Romei ◽  
Steven Boxer
Keyword(s):  
Rational Design ◽  
Fluorescent Protein ◽  
Fluorescent Proteins ◽  
Super Resolution ◽  
Unit Cell Size ◽  
Chlorine Substituent ◽  
Gfp Chromophore ◽  
The One ◽  
Green Fluorescent ◽  
Super Resolution Microscopy

<p>Double-bond photoisomerization in molecules such as the green fluorescent protein (GFP) chromophore can occur either via a volume-demanding one-bond-flip pathway or via a volume-conserving hula-twist pathway. Understanding the factors that determine the pathway of photoisomerization would inform the rational design of photoswitchable GFPs as improved tools for super-resolution microscopy. In this communication, we reveal the photoisomerization pathway of a photoswitchable GFP, rsEGFP2, by solving crystal structures of <i>cis</i> and <i>trans</i> rsEGFP2 containing a monochlorinated chromophore. The position of the chlorine substituent in the <i>trans</i> state breaks the symmetry of the phenolate ring of the chromophore and allows us to distinguish the two pathways. Surprisingly, we find that the pathway depends on the arrangement of protein monomers within the crystal lattice: in a looser packing, the one-bond-flip occurs, whereas in a tighter packing (7% smaller unit cell size), the hula-twist occurs.</p><p> </p><p> </p><p> </p><p> </p><p> </p><p> </p> <p> </p>

scholarly journals Advanced Static and Dynamic Fluorescence Microscopy Techniques to Investigate Drug Delivery Systems

Pharmaceutics ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 861
Author(s):  
Jacopo Cardellini ◽  
Arianna Balestri ◽  
Costanza Montis ◽  
Debora Berti
Keyword(s):  
Drug Delivery ◽  
Fluorescence Microscopy ◽  
Drug Delivery Systems ◽  
Laser Scanning ◽  
Super Resolution ◽  
Laser Scanning Confocal Microscopy ◽  
Delivery Systems ◽  
Scanning Confocal Microscopy ◽  
Biological Phenomena

In the past decade(s), fluorescence microscopy and laser scanning confocal microscopy (LSCM) have been widely employed to investigate biological and biomimetic systems for pharmaceutical applications, to determine the localization of drugs in tissues or entire organisms or the extent of their cellular uptake (in vitro). However, the diffraction limit of light, which limits the resolution to hundreds of nanometers, has for long time restricted the extent and quality of information and insight achievable through these techniques. The advent of super-resolution microscopic techniques, recognized with the 2014 Nobel prize in Chemistry, revolutionized the field thanks to the possibility to achieve nanometric resolution, i.e., the typical scale length of chemical and biological phenomena. Since then, fluorescence microscopy-related techniques have acquired renewed interest for the scientific community, both from the perspective of instrument/techniques development and from the perspective of the advanced scientific applications. In this contribution we will review the application of these techniques to the field of drug delivery, discussing how the latest advancements of static and dynamic methodologies have tremendously expanded the experimental opportunities for the characterization of drug delivery systems and for the understanding of their behaviour in biologically relevant environments.

A Multi-Functional Microfluidic Device Compatible with Widefield and Light Sheet Imaging

Lab on a Chip ◽  
2021 ◽  
Author(s):  
Regan P Moore ◽  
Ellen C O’Shaughnessy ◽  
Yu Shi ◽  
Ana T Nogueira ◽  
Katelyn M Heath ◽  
...  
Keyword(s):  
High Resolution ◽  
Microfluidic Device ◽  
Super Resolution ◽  
Light Sheet ◽  
Ethylene Propylene ◽  
Super Resolution Microscopy ◽  
Fluorinated Ethylene Propylene

We present a microfluidic device compatible with high resolution light sheet and super-resolution microscopy. Our device is a 150 μm thick chamber with a transparent fluorinated ethylene propylene (FEP) cover...

Cubic spline-based depth-dependent localization of mitochondria–endoplasmic reticulum contacts by three-dimensional light-sheet super-resolution microscopy

The Analyst ◽  
2021 ◽  
Author(s):  
Yucheng Sun ◽  
Seungah Lee ◽  
Seong Ho Kang
Keyword(s):  
Endoplasmic Reticulum ◽  
Calcium Homeostasis ◽  
Three Dimensional ◽  
Super Resolution ◽  
Light Sheet ◽  
Cubic Spline ◽  
Contact Distance ◽  
Super Resolution Microscopy

The contact distance between mitochondria (Mito) and endoplasmic reticulum (ER) has received considerable attention owing to their crucial function in maintaining lipid and calcium homeostasis. Herein, cubic spline algorithm-based depth-dependent...

mRNA quantification via second harmonic super resolution microscopy

2014 ◽  
Author(s):  
Jing Liu ◽  
Il-Hoon Cho ◽  
Ulhas Kadam ◽  
Joseph Irudayaraj
Keyword(s):  
Second Harmonic ◽  
Super Resolution ◽  
Mrna Quantification ◽  
Super Resolution Microscopy

Ultrasonically synthesized organic liquid-filled chitosan microcapsules: part 2: characterization using AFM (atomic force microscopy) and combined AFM–confocal laser scanning fluorescence microscopy

Soft Matter ◽  
2018 ◽  
Vol 14 (16) ◽  
pp. 3192-3201 ◽  
Author(s):  
Srinivas Mettu ◽  
Qianyu Ye ◽  
Meifang Zhou ◽  
Raymond Dagastine ◽  
Muthupandian Ashokkumar
Keyword(s):  
Atomic Force Microscopy ◽  
Fluorescence Microscopy ◽  
Young’S Modulus ◽  
Laser Scanning ◽  
Young's Modulus ◽  
Organic Liquid ◽  
Confocal Laser Scanning ◽  
Confocal Laser ◽  
Force Microscopy ◽  
Atomic Force

Atomic Force Microscopy (AFM) is used to measure the stiffness and Young's modulus of individual microcapsules that have a chitosan cross-linked shell encapsulating tetradecane.

Fluorescent Proteins and Super-Resolution Microscopy

2017 ◽  
Vol 37 (3) ◽  
pp. 0318008
Author(s):  
彭鼎铭 Peng Dingming ◽  
付志飞 Fu Zhifei ◽  
徐平勇 Xu Pingyong
Keyword(s):  
Fluorescent Proteins ◽  
Super Resolution ◽  
Super Resolution Microscopy

Novel dye for detection of callus embryo by confocal laser scanning fluorescence microscopy

Luminescence ◽  
2019 ◽  
Vol 34 (3) ◽  
pp. 353-359 ◽  
Author(s):  
Elena Kirilova ◽  
Ilona Mickevica ◽  
Ligita Mezaraupe ◽  
Aleksandrs Puckins ◽  
Ilze Rubenina ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Laser Scanning ◽  
Confocal Laser Scanning ◽  
Confocal Laser

Confocal Laser Scanning Fluorescence Microscopy

2013 ◽  
pp. 362-366
Author(s):  
Alberto Diaspro ◽  
Paolo Bianchini ◽  
Francesca Cella Zanacchi ◽  
Cesare Usai
Keyword(s):  
Fluorescence Microscopy ◽  
Laser Scanning ◽  
Confocal Laser Scanning ◽  
Confocal Laser
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