A green lifetime biosensor for calcium that remains bright over its full dynamic range DOI Open Access
Franka H. van der Linden, Stephen C. Thornquist,

Rick M ter Beek

et al.

Published: Jan. 2, 2025

Fluorescent biosensors toggle between two states and for the vast majority of one state is bright other dim. As a consequence, there substantial difference in signal-to-noise ratio (SNR) states. The dim has low ratio, which problematic when precise, quantitative measurements are needed. During engineering red-shifted variant an mTurquoise-based calcium sensor, we serendipitously generated green-emitting sensor that shows high brightness both calcium-bound -unbound state, while still showing dependent lifetime change >1 nanosecond. This named G-Ca-FLITS, comparable to GCaMP3 jGCaMP7c mammalian cells. induced loss fluorescence intensity only around 30% therefore observe little variation SNR levels change. G-Ca-FLITS negligible sensitivity pH physiological range, like its turquoise parent. Using imaging (FLIM), measured concentration with various organelles observed HeLa cells transient spatially heterogeneous elevations mitochondria. Finally, evaluated use predecessor two-photon FLIM Drosophila brains.

Language: Английский

Ultrasensitive sensors reveal the spatiotemporal landscape of lactate metabolism in physiology and disease DOI Creative Commons
Li Xie, Yinan Zhang, Lingyan Xu

et al.

Cell Metabolism, Journal Year: 2022, Volume and Issue: 35(1), P. 200 - 211.e9

Published: Oct. 28, 2022

Language: Английский

Citations

75
Lactate biosensors for spectrally and spatially multiplexed fluorescence imaging DOI Creative Commons
Yusuke Nasu, Abhi Aggarwal, Giang N. T. Le

et al.

Nature Communications, Journal Year: 2023, Volume and Issue: 14(1)

Published: Oct. 27, 2023

L-Lactate is increasingly appreciated as a key metabolite and signaling molecule in mammals. However, investigations of the inter- intra-cellular dynamics L-lactate are currently hampered by limited selection performance L-lactate-specific genetically encoded biosensors. Here we now report spectrally functionally orthogonal pair high-performance biosensors: green fluorescent extracellular biosensor, designated eLACCO2.1, red intracellular R-iLACCO1. eLACCO2.1 exhibits excellent membrane localization robust fluorescence response. To best our knowledge, R-iLACCO1 its affinity variants exhibit larger responses than any previously reported biosensor. We demonstrate spatially multiplexed imaging coexpression cultured cells, vivo mice.

Language: Английский

Citations

26
Next-Generation Genetically Encoded Fluorescent Biosensors Illuminate Cell Signaling and Metabolism DOI
Michelle S. Frei, Sohum Mehta, Jin Zhang

et al.

Annual Review of Biophysics, Journal Year: 2024, Volume and Issue: 53(1), P. 275 - 297

Published: Feb. 12, 2024

Genetically encoded fluorescent biosensors have revolutionized the study of cell signaling and metabolism, as they allow for live-cell measurements with high spatiotemporal resolution. This success has spurred development tailor-made that enable dynamic phenomena on different timescales length scales. In this review, we discuss approaches to enhancing developing new biosensors. We summarize technologies used gain structural insights into biosensor design comment useful screening technologies. Furthermore, give an overview applications where led key advances over recent years. Finally, our perspective future work is bound make a large impact.

Language: Английский

Citations

17
High-Performance Genetically Encoded Green Fluorescent Biosensors for Intracellular l-Lactate DOI Creative Commons
Saaya Hario, Giang N. T. Le, Hikaru Sugimoto

et al.

ACS Central Science, Journal Year: 2024, Volume and Issue: 10(2), P. 402 - 416

Published: Jan. 31, 2024

l-Lactate is a monocarboxylate produced during the process of cellular glycolysis and has long generally been considered waste product. However, studies in recent decades have provided new perspectives on physiological roles l-lactate as major energy substrate signaling molecule. To enable further investigations l-lactate, we developed series high-performance (ΔF/F = 15 to 30 vitro), intensiometric, genetically encoded green fluorescent protein (GFP)-based intracellular biosensors with range affinities. We evaluated these cultured cells demonstrated their application an ex vivo preparation Drosophila brain tissue. Using biosensors, were able detect glycolytic oscillations, which analyzed mathematically modeled.

Language: Английский

Citations

16
Comprehensive multiscale analysis of lactate metabolic dynamics in vitro and in vivo using highly responsive biosensors DOI
Aoxue Wang, Yejun Zou, Shuning Liu

et al.

Nature Protocols, Journal Year: 2024, Volume and Issue: 19(5), P. 1311 - 1347

Published: Feb. 2, 2024

Language: Английский

Citations

13
High-Sensitivity Detection of Tobacco Mosaic Virus RNA Based on a Ring-Opening Polymerization Fluorescent Biosensor DOI
Jing Yang, Jihong Chen, Yifan Li

et al.

Journal of Analysis and Testing, Journal Year: 2025, Volume and Issue: unknown

Published: March 19, 2025

Language: Английский

Citations

1
Fluorescence-Enhanced Microfluidic Biosensor Platform Based on Magnetic Beads with Highly Stable ZnO Nanorods for Biomarker Detection DOI
Montri Meeseepong, Gargi Ghosh, Sajal Shrivastava

et al.

ACS Applied Materials & Interfaces, Journal Year: 2023, Volume and Issue: 15(18), P. 21754 - 21765

Published: April 27, 2023

Existing affinity-based fluorescence biosensing systems for monitoring of biomarkers often utilize a fixed solid substrate immobilized with capture probes limiting their use in continuous or intermittent biomarker detection. Furthermore, there have been challenges integrating biosensors microfluidic chip and low-cost detector. Herein, we demonstrated highly efficient movable fluorescence-enhanced platform that can overcome the current limitations by combining enhancement digital imaging. Fluorescence-enhanced magnetic beads (MBs) decorated zinc oxide nanorods (MB-ZnO NRs) were used fluorescence-imaging-based aptasensing biomolecules improved signal-to-noise ratio. High stability homogeneous dispersion photostable MB-ZnO NRs obtained grafting bilayered silanes onto ZnO NRs. The formed on MB significantly signal up to 2.35 times compared without Moreover, integration device flow-based enabled measurements an electrolytic environment. results showed stable integrated significant potential diagnostics, biological assays, biomonitoring.

Language: Английский

Citations

20
Fluorescent Protein-Based Sensors for Detecting Essential Metal Ions across the Tree of Life DOI
Gary C. Jensen, Makena K. Janis, Hazel N. Nguyen

et al.

ACS Sensors, Journal Year: 2024, Volume and Issue: 9(4), P. 1622 - 1643

Published: April 8, 2024

Genetically encoded fluorescent metal ion sensors are powerful tools for elucidating dynamics in living systems. Over the last 25 years since first examples of genetically protein-based calcium indicators, this toolbox probes has expanded to include other essential and non-essential ions. Collectively, these have illuminated fundamental aspects homeostasis trafficking that crucial fields ranging from neurobiology human nutrition. Despite advances, much application remains limited mammalian cells tissues a number metals. Applications beyond systems vivo applications organisms primarily used sensors. The aim Perspective is provide, with support historical recent literature, an updated critical view design use detecting ions various organisms. We highlight progress achievements discuss more advances opportunities detection also outstanding challenges field directions future studies, including wider variety ions, developing implementing broader spectral range multiplexing experiments, applying single- multi-species biological

Language: Английский

Citations

9
Molecular Spies in Action: Genetically Encoded Fluorescent Biosensors Light up Cellular Signals DOI Creative Commons
Anneliese M. M. Gest, Ayse Z. Sahan, Yanghao Zhong

et al.

Chemical Reviews, Journal Year: 2024, Volume and Issue: 124(22), P. 12573 - 12660

Published: Nov. 13, 2024

Cellular function is controlled through intricate networks of signals, which lead to the myriad pathways governing cell fate. Fluorescent biosensors have enabled study these signaling in living systems across temporal and spatial scales. Over years there has been an explosion number fluorescent biosensors, as they become available for numerous targets, utilized spectral space, suited various imaging techniques. To guide users this extensive biosensor landscape, we discuss critical aspects proteins consideration development, smart tagging strategies, historical recent types, grouped by target, with a focus on design applications sensors systems.

Language: Английский

Citations

9
State-dependent motion of a genetically encoded fluorescent biosensor DOI Creative Commons
Paul C. Rosen, Samantha M. Horwitz, Daniel J. Brooks

et al.

Proceedings of the National Academy of Sciences, Journal Year: 2025, Volume and Issue: 122(10)

Published: March 6, 2025

Genetically encoded biosensors can measure biochemical properties such as small-molecule concentrations with single-cell resolution, even in vivo. Despite their utility, these sensors are "black boxes": Very little is known about the structures of low- and high-fluorescence states or what features required to transition between them. We used LiLac, a lactate biosensor quantitative fluorescence-lifetime readout, model system address questions. X-ray crystal engineered high-affinity metal bridges demonstrate that LiLac exhibits large interdomain twist motion pulls fluorescent protein away from "sealed," high-lifetime state absence "cracked," low-lifetime its presence. Understanding dynamics will help think engineer other biosensors.

Language: Английский

Citations

1