Design of a palette of SNAP-tag mimics of fluorescent proteins and their use as cell reporters DOI Creative Commons
Dasheng Zhang,

Zhengda Chen,

Zengmin Du

et al.

Cell Discovery, Journal Year: 2023, Volume and Issue: 9(1)

Published: June 13, 2023

Naturally occurring fluorescent proteins (FPs) are the most widely used tools for tracking cellular and sensing events. Here, we chemically evolved self-labeling SNAP-tag into a palette of mimics (SmFPs) that possess bright, rapidly inducible fluorescence ranging from cyan to infrared. SmFPs integral chemical-genetic entities based on same fluorogenic principle as FPs, i.e., induction non-emitting molecular rotors by conformational locking. We demonstrate usefulness these in real-time protein expression, degradation, binding interactions, trafficking, assembly, show optimally designed outperform FPs like GFP many important ways. further circularly permuted is sensitive changes their fusion partners, partners can be development single SmFP-based genetically encoded calcium sensors live cell imaging.

Language: Английский

Molecular Engineering of NIR‐II Fluorophores for Improved Biomedical Detection DOI
Zuhai Lei, Fan Zhang

Angewandte Chemie International Edition, Journal Year: 2020, Volume and Issue: 60(30), P. 16294 - 16308

Published: Aug. 12, 2020

The development of fluorophores for the second near-infrared window (NIR-II, 1000-1700 nm) represents an emerging, significant, and vibrant field in analytic chemistry, chemical biology, biomedical engineering. wavelength, brightness, stability are three crucial factors that determine performance NIR-II fluorophore. Up to now, significant progress has been made fluorescence molecular probes, including synthesis D-A-D D-π-A with improved imaging construction off-on probes ratiometric via energy transfer or structure modification. In this review, we summarize most recent advances engineering design strategies then highlight a selection bioimaging biosensing applications. We also provide perspectives on potential challenges opportunities emerging field.

Language: Английский

Citations

487

Rational Design of Fluorogenic and Spontaneously Blinking Labels for Super-Resolution Imaging DOI Creative Commons

Qinsi Zheng,

Anthony X. Ayala, Inhee Chung

et al.

ACS Central Science, Journal Year: 2019, Volume and Issue: 5(9), P. 1602 - 1613

Published: Sept. 5, 2019

Rhodamine dyes exist in equilibrium between a fluorescent zwitterion and nonfluorescent lactone. Tuning this toward the lactone form can improve cell-permeability allow creation of "fluorogenic" compounds-ligands that shift to upon binding biomolecular target. An archetype fluorogenic dye is far-red tetramethyl-Si-rhodamine (SiR), which has been used create exceptionally useful labels for advanced microscopy. Here, we develop quantitative framework development new dyes, determining lactone-zwitterion constant (KL-Z) sufficient predict fluorogenicity. This rubric emerged from our analysis known fluorophores yielded with improved performance cellular imaging experiments. We then designed novel fluorophore-Janelia Fluor 526 (JF526)-with SiR-like properties but shorter fluorescence excitation emission wavelengths. JF526 versatile scaffold probes including ligands self-labeling tags, stains endogenous structures, spontaneously blinking super-resolution immunofluorescence. constitutes label microscopy experiments, will enable rational design other bioimaging.

Language: Английский

Citations

215

Rational design of small molecule fluorescent probes for biological applications DOI
Joomyung V. Jun, David M. Chenoweth, E. James Petersson

et al.

Organic & Biomolecular Chemistry, Journal Year: 2020, Volume and Issue: 18(30), P. 5747 - 5763

Published: Jan. 1, 2020

Guidelines based on photophysical tuning, reactivity, isomerization, and energy transfer for rational design of synthetic fluorescent probes biological systems.

Language: Английский

Citations

196

Super-resolution Microscopy with Single Molecules in Biology and Beyond–Essentials, Current Trends, and Future Challenges DOI Creative Commons
Leonhard Möckl, W. E. Moerner

Journal of the American Chemical Society, Journal Year: 2020, Volume and Issue: 142(42), P. 17828 - 17844

Published: Oct. 9, 2020

Single-molecule super-resolution microscopy has developed from a specialized technique into one of the most versatile and powerful imaging methods nanoscale over past two decades. In this perspective, we provide brief overview historical development field, fundamental concepts, methodology required to obtain maximum quantitative information, current state art. Then, will discuss emerging perspectives areas where innovation further improvement are needed. Despite tremendous progress, full potential single-molecule is yet be realized, which enabled by research ahead us.

Language: Английский

Citations

152

Targeted multicolor in vivo imaging over 1,000 nm enabled by nonamethine cyanines DOI
Venugopal Bandi, Michael Luciano, Mara Saccomano

et al.

Nature Methods, Journal Year: 2022, Volume and Issue: 19(3), P. 353 - 358

Published: Feb. 28, 2022

Language: Английский

Citations

132

Advanced imaging and labelling methods to decipher brain cell organization and function DOI
Daniel Choquet, Matthieu Sainlos, Jean‐Baptiste Sibarita

et al.

Nature reviews. Neuroscience, Journal Year: 2021, Volume and Issue: 22(4), P. 237 - 255

Published: March 12, 2021

Language: Английский

Citations

93

Recent advances in near-infrared II imaging technology for biological detection DOI Creative Commons
Nannan Zhang,

Chenying Lu,

Min-jiang Chen

et al.

Journal of Nanobiotechnology, Journal Year: 2021, Volume and Issue: 19(1)

Published: May 10, 2021

Abstract Molecular imaging technology enables us to observe the physiological or pathological processes in living tissue at molecular level accurately diagnose diseases an early stage. Optical can be employed achieve dynamic monitoring of and has promising applications biomedicine. The traditional first near-infrared (NIR-I) window (NIR-I, range from 700 900 nm) technique been available for more than two decades extensively utilized clinical diagnosis, treatment scientific research. Compared with NIR-I, second NIR optical (NIR-II, 1000 1700 low autofluorescence, a high signal-to-noise ratio, penetration depth large Stokes shift. Recently, this attracted significant attention also become heavily researched topic In study, characteristics different fluorescence nanoprobes latest reports regarding application NIR-II biological tissues will described. Furthermore, existing problems future perspectives probes discussed.

Language: Английский

Citations

87

Defining the Basis of Cyanine Phototruncation Enables a New Approach to Single-Molecule Localization Microscopy DOI Creative Commons
Siddharth S. Matikonda,

Dominic A. Helmerich,

Mara Meub

et al.

ACS Central Science, Journal Year: 2021, Volume and Issue: 7(7), P. 1144 - 1155

Published: June 3, 2021

The light-promoted conversion of extensively used cyanine dyes to blue-shifted emissive products has been observed in various contexts. However, both the underlying mechanism and species involved this photoconversion reaction have remained elusive. Here we report that irradiation heptamethine cyanines provides pentamethine cyanines, which, turn, are photoconverted trimethine cyanines. We detail an examination substrate scope remarkable two-carbon phototruncation reaction. Supported by computational analysis, propose involves a singlet oxygen-initiated multistep sequence involving key hydroperoxycyclobutanol intermediate. Building on mechanistic framework, identify conditions improve yield over order magnitude. then demonstrate can be applied super-resolution single-molecule localization microscopy, leading improved spatial resolution with shorter imaging times. anticipate these insights will help transform common, but previously mechanistically ill-defined, chemical transformation into valuable optical tool.

Language: Английский

Citations

70

Single-molecule tracking of transcription protein dynamics in living cells: seeing is believing, but what are we seeing? DOI
Timothée Lionnet, Carl Wu

Current Opinion in Genetics & Development, Journal Year: 2021, Volume and Issue: 67, P. 94 - 102

Published: Jan. 9, 2021

Language: Английский

Citations

61

Photochemical Mechanisms of Fluorophores Employed in Single‐Molecule Localization Microscopy DOI Creative Commons
Kai Kikuchi, Liam D. Adair, Jiarun Lin

et al.

Angewandte Chemie International Edition, Journal Year: 2022, Volume and Issue: 62(1)

Published: Sept. 30, 2022

Decoding cellular processes requires visualization of the spatial distribution and dynamic interactions biomolecules. It is therefore not surprising that innovations in imaging technologies have facilitated advances biomedical research. The advent super-resolution has empowered researchers with ability to answer long-standing questions about at an entirely new level. Fluorescent probes greatly enhance specificity resolution experiments. Here, we introduce key technologies, a brief discussion on single-molecule localization microscopy (SMLM). We evaluate chemistry photochemical mechanisms fluorescent employed SMLM. This Review provides guidance identification adoption single molecule inspire design next-generation amenable imaging.

Language: Английский

Citations

39