The promise and peril of comparing fluorescence lifetime in biology revealed by simulations DOI Open Access
Pingchuan Ma,

Scott Sternson,

Yao Chen

et al.

Published: Nov. 12, 2024

Signaling dynamics are crucial in biological systems, and biosensor-based real-time imaging has revolutionized their analysis. Fluorescence lifetime microscopy (FLIM) excels over the widely used fluorescence intensity by allowing measurement of absolute signal levels, independent sensor concentration. This capability enables comparison signaling across different animals, body regions, timeframes. However, FLIM’s advantage can be compromised factors like autofluorescence experiments. To address this, we introduce FLiSimBA, a flexible computational framework for realistic F luorescence Li fetime Sim ulation B iological A pplications. Through simulations, analyze signal-to-noise ratios data, determining uncertainty providing necessary error bars measurements. Furthermore, challenge belief that is unaffected expression establish quantitative limits to this insensitivity applications. Additionally, propose innovations, notably multiplexed dynamic combines innovation transform number signals simultaneously monitored, thereby enabling systems approach studying dynamics. Thus, incorporating diverse into our simulation framework, uncover surprises, identify limitations, advancements biology. supports rigorous experimental design, facilitates accurate data interpretation, paves way technological imaging.

Language: Английский

Absolute measurement of fast and slow neuronal signals with fluorescence lifetime photometry at high temporal resolution DOI Creative Commons

Bart Lodder,

Tarun V. Kamath,

Ecaterina Savenco

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 12, 2025

The concentrations of extracellular and intracellular signaling molecules, such as dopamine cAMP, change over both fast slow timescales impact downstream pathways in a cell-type specific manner. Fluorescence sensors currently used to monitor signals vivo are typically optimized detect fast, relative changes concentration the target molecule. They less well suited slowly-changing rarely provide absolute measurements either components. Here, we developed system for fluorescence lifetime photometry at high temporal resolution (FLIPR) that utilizes frequency-domain analog processing measure genetically-encoded speed but with long-term stability picosecond precision freely moving mice. We applied FLIPR investigate two functionally distinct regions striatum, nucleus accumbens core (NAC) tail striatum (TOS). observed higher tonic levels baseline TOS compared NAC detected differential dynamic responses phasic appetitive aversive stimuli. Thus, enables simple monitoring time-scale neuronal units, revealing previously unappreciated spatial variation even well-studied systems.

Language: Английский

Citations

3

A modular chemigenetic calcium indicator for multiplexed in vivo functional imaging DOI Creative Commons
Helen Farrants, Yichun Shuai, William C. Lemon

et al.

Nature Methods, Journal Year: 2024, Volume and Issue: 21(10), P. 1916 - 1925

Published: Sept. 20, 2024

Abstract Genetically encoded fluorescent calcium indicators allow cellular-resolution recording of physiology. However, bright, genetically targetable that can be multiplexed with existing tools in vivo are needed for simultaneous imaging multiple signals. Here we describe WHaloCaMP, a modular chemigenetic indicator built from bright dye-ligands and protein sensor domains. Fluorescence change WHaloCaMP results reversible quenching the bound dye via strategically placed tryptophan. is compatible rhodamine fluoresce green to near-infrared, including several efficiently label brain animals. When near-infrared dye-ligand, shows 7× increase fluorescence intensity 2.1-ns lifetime upon binding. We use WHaloCaMP1a image Ca 2+ responses flies mice, perform three-color functional hundreds neurons astrocytes zebrafish larvae quantify concentration using microscopy (FLIM).

Language: Английский

Citations

11

A Cre-dependent reporter mouse for quantitative real-time imaging of protein kinase A activity dynamics DOI Creative Commons

Elizabeth I. Tilden,

Aditi Maduskar,

Anna Oldenborg

et al.

Scientific Reports, Journal Year: 2024, Volume and Issue: 14(1)

Published: Feb. 6, 2024

Intracellular signaling dynamics play a crucial role in cell function. Protein kinase A (PKA) is key molecule that has diverse functions, from regulating metabolism and brain activity to guiding development cancer progression. We previously developed an optical reporter, FLIM-AKAR, allows for quantitative imaging of PKA via fluorescence lifetime microscopy photometry. However, using viral infection or electroporation the delivery FLIM-AKAR invasive results variable expression. Here, we reporter mouse, FL-AK, which expresses Cre-dependent manner ROSA26 locus. FL-AK provides robust consistent expression over time. Functionally, mouse line reports increase response activation both G

Language: Английский

Citations

6

Serotonin signaling at cilia synapses DOI Creative Commons
Katherine DeLong, Shu‐Hsien Sheu

Current Opinion in Neurobiology, Journal Year: 2025, Volume and Issue: 92, P. 102994 - 102994

Published: March 12, 2025

Language: Английский

Citations

0

Quantification of extracellular glutamate levels using fluorescence lifetime imaging of the red genetically encoded biosensor Rncp-iGluSnFR1 DOI
Huangmei Zhou, Lin Wu, Weichun Jiang

et al.

Microchemical Journal, Journal Year: 2025, Volume and Issue: 212, P. 113571 - 113571

Published: April 10, 2025

Language: Английский

Citations

0

An integrated microfluidic and fluorescence platform for probing in vivo neuropharmacology DOI
Sean C. Piantadosi, Min‐Kyu Lee, Mingzheng Wu

et al.

Neuron, Journal Year: 2025, Volume and Issue: unknown

Published: April 1, 2025

Language: Английский

Citations

0

Fiber photometry in neuroscience research: principles, applications, and future directions DOI Creative Commons
Michał Kiełbiński, Joanna Bernacka

Pharmacological Reports, Journal Year: 2024, Volume and Issue: unknown

Published: Sept. 5, 2024

Language: Английский

Citations

1

The promise and peril of comparing fluorescence lifetime in biology revealed by simulations DOI Open Access
Pingchuan Ma,

Scott Sternson,

Yao Chen

et al.

Published: Nov. 12, 2024

Signaling dynamics are crucial in biological systems, and biosensor-based real-time imaging has revolutionized their analysis. Fluorescence lifetime microscopy (FLIM) excels over the widely used fluorescence intensity by allowing measurement of absolute signal levels, independent sensor concentration. This capability enables comparison signaling across different animals, body regions, timeframes. However, FLIM’s advantage can be compromised factors like autofluorescence experiments. To address this, we introduce FLiSimBA, a flexible computational framework for realistic F luorescence Li fetime Sim ulation B iological A pplications. Through simulations, analyze signal-to-noise ratios data, determining uncertainty providing necessary error bars measurements. Furthermore, challenge belief that is unaffected expression establish quantitative limits to this insensitivity applications. Additionally, propose innovations, notably multiplexed dynamic combines innovation transform number signals simultaneously monitored, thereby enabling systems approach studying dynamics. Thus, incorporating diverse into our simulation framework, uncover surprises, identify limitations, advancements biology. supports rigorous experimental design, facilitates accurate data interpretation, paves way technological imaging.

Language: Английский

Citations

1

Citrus Polymethoxyflavones Regulate against Aging-Associated Diseases: Advances in Biological Mechanisms Responsible for Their Regulation DOI
Xiaojuan Chen,

Weiyi Sun,

Sicheng Ji

et al.

Journal of Agricultural and Food Chemistry, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 11, 2024

As the proportion of aging population globally is surging year by year, age-associated diseases, including neurodegenerative, metabolic, and cardiovascular have recently attracted widespread attention food scientists nutritionists. Polymethoxyflavonoids (PMFs), a type dietary flavonoids, emerged as potential antiaging candidates owing to their diverse bioactivities, encompassing antioxidant, anti-inflammatory, neuroprotective, metabolic regulatory effects. Herein, this comprehensive updated review has summarized discussed effects PMFs on aging, possible mechanisms that link PMFs-mediated modulation prevention or treatment various aging-related diseases been elaborated in detail. Furthermore, biological fate elaborately from absorption, distribution, metabolism, excretion vivo. Special given bioavailability-bioactivity relationship PMFs, PMF's activity significantly hampered poor bioavailability. Overall, all these conclusions may help providing perspective for further study aging.

Language: Английский

Citations

1

The promise and peril of comparing fluorescence lifetime in biology revealed by simulations DOI Open Access
Pingchuan Ma,

Scott Sternson,

Yao Chen

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Dec. 21, 2023

Abstract Signaling dynamics are crucial in biological systems, and biosensor-based real-time imaging has revolutionized their analysis. Fluorescence lifetime microscopy (FLIM) excels over the widely used fluorescence intensity by allowing measurement of absolute signal levels, independent sensor concentration. This capability enables comparison signaling across different animals, body regions, timeframes. However, FLIM’s advantage can be compromised factors like autofluorescence experiments. To address this, we introduce FLiSimBA, a flexible computational framework for realistic F luorescence Li fetime Sim ulation B iological A pplications. Through simulations, analyze signal-to-noise ratios data, determining uncertainty providing necessary error bars measurements. Furthermore, challenge belief that is unaffected expression establish quantitative limits to this insensitivity applications. Additionally, propose innovations, notably multiplexed dynamic combines innovation transform number signals simultaneously monitored, thereby enabling systems approach studying dynamics. Thus, incorporating diverse into our simulation framework, uncover surprises, identify limitations, advancements biology. supports rigorous experimental design, facilitates accurate data interpretation, paves way technological imaging.

Language: Английский

Citations

2