Benefits of Combined Fluorescence Lifetime Imaging Microscopy and Fluorescence Correlation Spectroscopy for Biomedical Studies Demonstrated by Using a Liposome Model System DOI Creative Commons

Kristina Bruun,

Hans‐Gerd Löhmannsröben, Carsten Hille

et al.

Biophysica, Journal Year: 2024, Volume and Issue: 4(2), P. 207 - 226

Published: April 25, 2024

Drug delivery systems play a pivotal role in targeted pharmaceutical transport and controlled release at specific sites. Liposomes, commonly used as drug carriers, constitute fundamental part of these systems. Moreover, the drug–liposome model serves robust platform for investigating interaction processes both cellular molecular levels. To advance our understanding carrier uptake mechanisms, we employed fluorescence lifetime imaging microscopy (FLIM) correlation spectroscopy (FCS), leveraging unique benefits two-photon (2P) excitation. Our approach utilized giant unilamellar vesicles (GUVs) simplified system cell membranes, labelled with amphiphilic fluorescent dye 3,3′-dioctadecyloxa-carbocyanine (DiOC18(3)). Additionally, large (LUVs) functioned system, incorporating spectrally distinct sulforhodamine 101 (SRh101) surrogate drug. The investigation emphasized diverse interactions between GUVs LUVs based on charged lipids employed. We examined exchange kinetics structural alterations liposome carriers during process. study underscores significance employing 2P excitation conjunction FLIM FCS. This powerful combination offers valuable methodological studying interactions, positioning them an exceptionally versatile technical advantage.

Language: Английский

Fluorescence lifetime imaging in drug delivery research DOI Creative Commons
Yiqing Lu,

Parinaz Jabbari,

Anton Mukhamedshin

et al.

Advanced Drug Delivery Reviews, Journal Year: 2025, Volume and Issue: 218, P. 115521 - 115521

Published: Jan. 22, 2025

Language: Английский

Citations

2
Development of time-resolved luminescence measurement instruments for biosensing and bioimaging – An overview DOI
Benjamin Sreenan, Vala Kafil,

Donovan Wells

et al.

Measurement, Journal Year: 2025, Volume and Issue: unknown, P. 117201 - 117201

Published: March 1, 2025

Language: Английский

Citations

1
Spectroscopic properties of red-emitting Sr0.9Na0.2ZrO3:Eu3+ phosphors with potential applications in lasers and LEDs DOI
A. Nathan‐Abutu, I. Ahemen, R.E. Kroon

et al.

Optical Materials, Journal Year: 2024, Volume and Issue: unknown, P. 116338 - 116338

Published: Oct. 1, 2024

Language: Английский

Citations

4
The optimization and application of photodynamic diagnosis and autofluorescence imaging in tumor diagnosis and guided surgery: current status and future prospects DOI Creative Commons
Wei Keat Wan, Huiquan Liu, Junrong Zou

et al.

Frontiers in Oncology, Journal Year: 2025, Volume and Issue: 14

Published: Jan. 8, 2025

Photodynamic diagnosis (PDD) and autofluorescence imaging (AFI) are emerging cancer diagnostic technologies that offer significant advantages over traditional white-light endoscopy in detecting precancerous lesions early-stage cancers; moreover, they hold promising potential fluorescence-guided surgery (FGS) for tumors. However, their shortcomings have somewhat hindered the clinical application of PDD AFI. Therefore, it is imperative to enhance efficacy AFI, thereby maximizing practical use. This article reviews principles, characteristics, current research status, advancements focusing on analyzing discussing optimization strategies AFI tumor FGS scenarios. Considering technical feasibility, optimizing may result an effective real-time tool guide clinicians surgical guidance achieve best results.

Language: Английский

Citations

0
Rapid Acquisition of High-Pixel Fluorescence Lifetime Images of Living Cells via Image Reconstruction Based on Edge-Preserving Interpolation DOI Creative Commons

Yinru Zhu,

Yong Guo,

Xinwei Gao

et al.

Biosensors, Journal Year: 2025, Volume and Issue: 15(1), P. 43 - 43

Published: Jan. 13, 2025

Fluorescence lifetime imaging (FLIM) has established itself as a pivotal tool for investigating biological processes within living cells. However, the extensive duration necessary to accumulate sufficient photons accurate fluorescence calculations poses significant obstacle achieving high-resolution monitoring of cellular dynamics. In this study, we introduce an image reconstruction method based on edge-preserving interpolation (EPIM), which transforms rapidly acquired low-resolution FLIM data into high-pixel images, thereby eliminating need extended acquisition times. Specifically, decouple grayscale and matrix perform individual each. Following intensity image, apply wavelet transformation adjust coefficients according gradients. After inverse transformation, original is obtained subjected noise reduction complete process. Subsequently, each pixel pseudo-color-coded its lifetime, preserving both structural temporal information. We evaluated performance bicubic our approach microspheres fixed-cell samples, demonstrating their effectiveness in enhancing quality images. By applying these techniques live-cell imaging, can successfully obtain images at shortened intervals, facilitating capture rapid events.

Language: Английский

Citations

0
Label-free metabolic fingerprinting of motile mammalian spermatozoa with subcellular resolution DOI Creative Commons
Fitore Kusari,

Lenka Backová,

Dalibor Pánek

et al.

BMC Biology, Journal Year: 2025, Volume and Issue: 23(1)

Published: March 24, 2025

Sperm metabolic pathways that generate energy for motility are compartmentalized within the flagellum. Dysfunctions in compartments, namely mitochondrial respiration and glycolysis, can compromise male fertility. Studying these compartments is thus required fertility treatment. However, it very challenging to capture images of motile spermatozoa because fast beating flagellum introduces motion blur. Therefore, most approaches immobilize prior imaging. Our findings indicate immobilizing sperm alters their profile, highlighting necessity measuring metabolism during movement. We achieved this by encapsulating mouse epididymis a hydrogel followed two-photon fluorescence lifetime imaging microscopy situ. The autofluorescence endogenous metabolites—FAD, NADH, NADPH—enabled us visualize without staining. trained machine learning automated image segmentation generated fingerprints using object-based phasor analysis. show compartment (1) distinguish individual males genetic background, age, or fecundity status, (2) correlate with fertility, (3) change age likely due increased oxidative metabolism. approach eliminates need immobilization labeling captures native state This technique could be adapted metabolism-based selection assisted reproduction.

Language: Английский

Citations

0
Modeling Tumor Microenvironment Complexity In Vitro: Spheroids as Physiologically Relevant Tumor Models and Strategies for Their Analysis DOI Creative Commons

Shrey Shah,

Gerard G. M. D’Souza

Cells, Journal Year: 2025, Volume and Issue: 14(10), P. 732 - 732

Published: May 17, 2025

Drug delivery to solid tumors is challenged by multiple physiological barriers arising from the tumor microenvironment, including dense extracellular matrix, cellular heterogeneity, hypoxic gradients, and elevated interstitial fluid pressure. These features hinder uniform distribution accumulation of therapeutics, reducing treatment efficacy. Despite their widespread use, conventional two-dimensional monolayer cultures fail reproduce these complexities, contributing poor translational predictability many preclinical candidates. Three-dimensional multicellular spheroids have emerged as more representative in vitro models that capture essential architecture, stromal interactions, microenvironmental resistance mechanisms. Spheroids exhibit spatially organized regions proliferation, quiescence, hypoxia, can incorporate non-tumor cells mimic tumor–stroma crosstalk. Advances spheroid analysis now enable detailed evaluation drug penetration, migration, cytotoxic response, molecular gradients using techniques such optical confocal imaging, large-particle flow cytometry, biochemical viability assays, microfluidic integration. By combining relevance with analytical accessibility, support mechanistic studies transport efficacy under tumor-like conditions. Their adoption into routine workflows has potential improve accuracy while reliance on animal models.

Language: Английский

Citations

0
Characterization of homogeneous energy transfer kinetics by time-correlated single photon counting DOI
Yongjia Wang, Di Tian, Zece Zhu

et al.

Journal of Photochemistry and Photobiology A Chemistry, Journal Year: 2025, Volume and Issue: unknown, P. 116514 - 116514

Published: May 1, 2025

Language: Английский

Citations

0
Fluorescence Lifetime Imaging Microscopy: Advances in Materials Science Research DOI
Fariyad Ali, Subhankar Kundu

Chemistry of Materials, Journal Year: 2025, Volume and Issue: unknown

Published: May 27, 2025

Language: Английский

Citations

0
Neural Metabolic Networks: Key Elements of Healthy Brain Function DOI Creative Commons
Nimrod Madrer, Nirma D. Perera, Nonthué A. Uccelli

et al.

Journal of Neurochemistry, Journal Year: 2025, Volume and Issue: 169(6)

Published: June 1, 2025

Neural networks are responsible for processing sensory stimuli and driving the synaptic activity required brain function behavior. This computational capacity is expensive requires a steady supply of energy building blocks to operate. Importantly, neural composed different cell populations, whose metabolic profiles differ between each other, thus endowing them with capacities, such as, example, ability synthesize specific precursors or variable proficiency manage their waste. These marked differences likely prompted emergence diverse intercellular interactions, in which shuttling cycling metabolites cells allows separation workload efficient control demand within central nervous system. Nevertheless, our knowledge about bioenergetics adaptations still warrants further studies. In this review, originated from Fourth International Society Neurochemistry (ISN) Journal (JNC) Flagship School held Schmerlenbach, Germany (2022), we describe discuss cells, exchanges these how bioenergetic activities shape Furthermore, potential role faulty etiology progression Alzheimer's disease, Parkinson Amyotrophic lateral sclerosis. We foresee that deeper understanding metabolism will provide crucial insights into higher-order functions emerge reveal roots neuropathological conditions hallmarks include impaired function.

Language: Английский

Citations

0