scholarly journals Red fluorescent proteins (RFPs) and RFP-based biosensors for neuronal imaging applications

2015 ◽  
Vol 2 (3) ◽  
pp. 031203 ◽  
Author(s):  
Yi Shen ◽  
Tiffany Lai ◽  
Robert E. Campbell
Keyword(s):  
Fluorescent Proteins ◽  
Red Fluorescent Proteins ◽  
Neuronal Imaging

POSSIBLE APPLICATION OF METAL SENSITIVE RED FLUORESCENT PROTEINS IN ENVIRONMENTAL MONITORING

2012 ◽  
Vol 11 (1) ◽  
pp. 193-198 ◽  
Author(s):  
Laszlo Szilagyi ◽  
Maria Szabo (Palfi) ◽  
Judit Petres ◽  
Ildiko Miklossy ◽  
Beata Abraham ◽  
...  
Keyword(s):  
Environmental Monitoring ◽  
Fluorescent Proteins ◽  
Red Fluorescent Proteins

scholarly journals Split-wrmScarlet and split-sfGFP: Tools for faster, easier fluorescent l abeling of endogenous proteins in Caenorhabditis elegans

Genetics ◽  
2021 ◽  
Author(s):  
Jérôme Goudeau ◽  
Catherine S Sharp ◽  
Jonathan Paw ◽  
Laura Savy ◽  
Manuel D Leonetti ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Fluorescent Proteins ◽  
Fluorescent Labeling ◽  
Large Fragment ◽  
C Elegans ◽  
High Affinity ◽  
Red Fluorescent Proteins ◽  
Invaluable Tool ◽  
Endogenous Proteins ◽  
Fluorescent Tags

Abstract We create and share a new red fluorophore, along with a set of strains, reagents and protocols, to make it faster and easier to label endogenous C. elegans proteins with fluorescent tags. CRISPR-mediated fluorescent labeling of C. elegans proteins is an invaluable tool, but it is much more difficult to insert fluorophore-size DNA segments than it is to make small gene edits. In principle, high-affinity asymmetrically split fluorescent proteins solve this problem in C. elegans: the small fragment can quickly and easily be fused to almost any protein of interest, and can be detected wherever the large fragment is expressed and complemented. However, there is currently only one available strain stably expressing the large fragment of a split fluorescent protein, restricting this solution to a single tissue (the germline) in the highly autofluorescent green channel. No available C. elegans lines express unbound large fragments of split red fluorescent proteins, and even state-of-the-art split red fluorescent proteins are dim compared to the canonical split-sfGFP protein. In this study, we engineer a bright, high-affinity new split red fluorophore, split-wrmScarlet. We generate transgenic C. elegans lines to allow easy single-color labeling in muscle or germline cells and dual-color labeling in somatic cells. We also describe a novel expression strategy for the germline, where traditional expression strategies struggle. We validate these strains by targeting split-wrmScarlet to several genes whose products label distinct organelles, and we provide a protocol for easy, cloning-free CRISPR/Cas9 editing. As the collection of split-FP strains for labeling in different tissues or organelles expands, we will post updates at doi.org/10.5281/zenodo.3993663

Generation and Characterization of a Rat Monoclonal Antibody Specific for Multiple Red Fluorescent Proteins

Hybridoma ◽  
2008 ◽  
Vol 27 (5) ◽  
pp. 337-343 ◽  
Author(s):  
Andrea Rottach ◽  
Elisabeth Kremmer ◽  
Danny Nowak ◽  
Heinrich Leonhardt ◽  
M. Cristina Cardoso
Keyword(s):  
Monoclonal Antibody ◽  
Fluorescent Proteins ◽  
Red Fluorescent Proteins

scholarly journals Monomerization of far-red fluorescent proteins

2018 ◽  
Vol 115 (48) ◽  
pp. E11294-E11301 ◽  
Author(s):  
Timothy M. Wannier ◽  
Sarah K. Gillespie ◽  
Nicholas Hutchins ◽  
R. Scott McIsaac ◽  
Sheng-Yi Wu ◽  
...  
Keyword(s):  
Molecular Markers ◽  
Biological Markers ◽  
Fluorescent Proteins ◽  
Biological Tissues ◽  
Biological Applications ◽  
Emission Maximum ◽  
Naturally Occurring ◽  
Red Fluorescent Proteins ◽  
Animal Imaging ◽  
Structural Perturbations

Anthozoa-class red fluorescent proteins (RFPs) are frequently used as biological markers, with far-red (λem ∼ 600–700 nm) emitting variants sought for whole-animal imaging because biological tissues are more permeable to light in this range. A barrier to the use of naturally occurring RFP variants as molecular markers is that all are tetrameric, which is not ideal for cell biological applications. Efforts to engineer monomeric RFPs have typically produced dimmer and blue-shifted variants because the chromophore is sensitive to small structural perturbations. In fact, despite much effort, only four native RFPs have been successfully monomerized, leaving the majority of RFP biodiversity untapped in biomarker development. Here we report the generation of monomeric variants of HcRed and mCardinal, both far-red dimers, and describe a comprehensive methodology for the monomerization of red-shifted oligomeric RFPs. Among the resultant variants is mKelly1 (emission maximum, λem = 656 nm), which, along with the recently reported mGarnet2 [Matela G, et al. (2017) Chem Commun (Camb) 53:979–982], forms a class of bright, monomeric, far-red FPs.

scholarly journals Quantitative Determination of Dark Chromophore Population Explains the Apparent Low Quantum Yield of Red Fluorescent Proteins

2020 ◽  
Vol 124 (8) ◽  
pp. 1383-1391 ◽  
Author(s):  
Jord C. Prangsma ◽  
Robert Molenaar ◽  
Laura van Weeren ◽  
Daphne S. Bindels ◽  
Lindsay Haarbosch ◽  
...  
Keyword(s):  
Quantum Yield ◽  
Quantitative Determination ◽  
Fluorescent Proteins ◽  
Red Fluorescent Proteins

A General Mechanism of Photoconversion of Green-to-Red Fluorescent Proteins Based on Blue and Infrared Light Reduces Phototoxicity in Live-Cell Single-Molecule Imaging

2017 ◽  
Vol 56 (38) ◽  
pp. 11634-11639 ◽  
Author(s):  
Bartosz Turkowyd ◽  
Alexander Balinovic ◽  
David Virant ◽  
Haruko G. Gölz Carnero ◽  
Fabienne Caldana ◽  
...  
Keyword(s):  
Single Molecule ◽  
Fluorescent Proteins ◽  
Live Cell ◽  
Infrared Light ◽  
General Mechanism ◽  
Single Molecule Imaging ◽  
Red Fluorescent Proteins

scholarly journals Multiphoton Bleaching of Red Fluorescent Proteins and the Ways to Reduce It

2022 ◽  
Vol 23 (2) ◽  
pp. 770
Author(s):  
Mikhail Drobizhev ◽  
Rosana S. Molina ◽  
Jacob Franklin
Keyword(s):  
Chemical Structure ◽  
Fluorescent Proteins ◽  
Multiphoton Ionization ◽  
Excitation Power ◽  
Photon Absorption ◽  
Green Fluorescent Proteins ◽  
Two Photon Absorption ◽  
Two Photon ◽  
Red Fluorescent Proteins ◽  
Green Fluorescent

Red fluorescent proteins and biosensors built upon them are potentially beneficial for two-photon laser microscopy (TPLM) because they can image deeper layers of tissue, compared to green fluorescent proteins. However, some publications report on their very fast photobleaching, especially upon excitation at 750–800 nm. Here we study the multiphoton bleaching properties of mCherry, mPlum, tdTomato, and jREX-GECO1, measuring power dependences of photobleaching rates K at different excitation wavelengths across the whole two-photon absorption spectrum. Although all these proteins contain the chromophore with the same chemical structure, the mechanisms of their multiphoton bleaching are different. The number of photons required to initiate a photochemical reaction varies, depending on wavelength and power, from 2 (all four proteins) to 3 (jREX-GECO1) to 4 (mCherry, mPlum, tdTomato), and even up to 8 (tdTomato). We found that at sufficiently low excitation power P, the rate K often follows a quadratic power dependence, that turns into higher order dependence (K~Pα with α > 2) when the power surpasses a particular threshold P*. An optimum intensity for TPLM is close to the P*, because it provides the highest signal-to-background ratio and any further reduction of laser intensity would not improve the fluorescence/bleaching rate ratio. Additionally, one should avoid using wavelengths shorter than a particular threshold to avoid fast bleaching due to multiphoton ionization.

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