Förster resonance energy transfer (FRET) is a powerful biological tool for reading out cell signaling processes. In vivo use of FRET is challenging because of the scattering properties of bulk tissue. By combining diffuse fluorescence tomography with fluorescence lifetime imaging (FLIM), implemented using wide-field time-gated detection of fluorescence excited by ultrashort laser pulses in a tomographic imaging system and applying inverse scattering algorithms, we can reconstruct the three dimensional spatial localization of fluorescence quantum efficiency and lifetime. We demonstrate in vivo spatial mapping of FRET between genetically expressed fluorescent proteins in live mice read out using FLIM. Following transfection by electroporation, mouse hind leg muscles were imaged in vivo and the emission of free donor (eGFP) in the presence of free acceptor (mCherry) could be clearly distinguished from the fluorescence of the donor when directly linked to the acceptor in a tandem (eGFP-mCherry) FRET construct.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3130577 | PMC |
| http://dx.doi.org/10.1364/BOE.2.001907 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Fluorescence Lifetime Endoscopy with a Nanosecond Time-Gated CAPS Camera with IRF-Free Deep Learning Method.
Sensors (Basel)
January 2025
Department of Electronics and Informatics (ETRO), Vrije Universiteit Brussel, 1050 Brussels, Belgium.
Fluorescence imaging has been widely used in fields like (pre)clinical imaging and other domains. With advancements in imaging technology and new fluorescent labels, fluorescence lifetime imaging is gradually gaining recognition. Our research department is developing the CAM, based on the Current-Assisted Photonic Sampler, to achieve real-time fluorescence lifetime imaging in the NIR (700-900 nm) region.
View Article and Find Full Text PDFLuminescent Lanthanide Infinite Coordination Polymers for Ratiometric Sensing Applications.
Molecules
January 2025
College of Chemistry and Chemical Engineering, Central South University, Changsha 410017, China.
Ratiometric lanthanide coordination polymers (Ln-CPs) are advanced materials that combine the unique optical properties of lanthanide ions (e.g., Eu, Tb, Ce) with the structural flexibility and tunability of coordination polymers.
View Article and Find Full Text PDFCoordination Self-Assembly Induced Hot Exciton Fluorescence and Multi-Source Excitation Long Afterglow.
Angew Chem Int Ed Engl
January 2025
Sun Yat-Sen University, School of Chemistry and Chemical Engineering, 135 West Xingang Road, 510275, Guangzhou, CHINA.
Metal-organic complexes with long afterglow luminescence have attracted extensive attention due to potential applications in display, sensing and information security. However, most of the metal-organic complex long afterglow materials reported so far are limited to the use of UV light as the excitation source, and the ambiguity of the structure-activity relationship makes the development of metal-organic complexes extremely limited. Herein, a series of metal-organic complexes with ultralong emission lifetime is constructed by coordination assembly of Zn(II) with three isomers.
View Article and Find Full Text PDFPreferential Binding of a Long-Arm Porphyrin Ligand to Higher Order G-Quadruplex under Crowding Conditions.
J Phys Chem B
January 2025
College of Chemistry, Beijing Normal University, Beijing 100875, P. R. China.
Under conditions that are close to the real cellular environment, the human telomeric single-stranded overhang (∼200 nt) consisting of tens of TTAGGG repeats tends to form higher order structures of multiple G-quadruplex (G4) blocks. On account of the higher biological relevance of higher order G4 structures, ligand compounds binding to higher order G4 are significant for the drug design toward inhibiting telomerase activity. Here, we study the interaction between a cationic porphyrin derivative, 5,10,15,20-tetra{4-[2-(1-methyl-1-piperidinyl)propoxy]phenyl}porphyrin (T4), and a human telomeric G4-dimer (AG(TAG)) in the mimic intracellular molecularly crowded environment (PEG as a crowding agent) and K or Na solution (i.
View Article and Find Full Text PDFRapid Acquisition of High-Pixel Fluorescence Lifetime Images of Living Cells via Image Reconstruction Based on Edge-Preserving Interpolation.
Biosensors (Basel)
January 2025
State Key Laboratory of Radio Frequency Heterogeneous Integration (Shenzhen University), Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen 518060, China.
Fluorescence lifetime imaging (FLIM) has established itself as a pivotal tool for investigating biological processes within living cells. However, the extensive imaging duration necessary to accumulate sufficient photons for accurate fluorescence lifetime calculations poses a significant obstacle to achieving high-resolution monitoring of cellular dynamics. In this study, we introduce an image reconstruction method based on the edge-preserving interpolation method (EPIM), which transforms rapidly acquired low-resolution FLIM data into high-pixel images, thereby eliminating the need for extended acquisition times.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!