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fluorescence lifetime imaging microscopy

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https://www.readbyqxmd.com/read/28985028/pitfalls-in-using-fluorescence-tagging-of-nanomaterials-tecto-dendrimers-in-skin-tissue-as-investigated-by-cluster-flim
#1
Pierre Volz, Priscila Schilrreff, Robert Brodwolf, Christopher Wolff, Johannes Stellmacher, Jens Balke, Maria J Morilla, Christian Zoschke, Monika Schäfer-Korting, Ulrike Alexiev
Targeted topical application promises high drug concentrations in the skin and low systemic adverse effects. To locate drugs and drug-delivery systems like nanocarriers, fluorescent dyes are commonly used as drug surrogates or nanocarrier labels in micrographs of tissue sections. Here, we investigate how labeling degree, concentration of fluorophore, and nanocarrier may affect the interpretation of these micrographs. False-negative penetration results due to inter- and intramolecular quenching effects are likely...
October 2017: Annals of the New York Academy of Sciences
https://www.readbyqxmd.com/read/28984329/a-water-soluble-two-photon-ratiometric-triarylboron-probe-with-nucleolar-targeting-by-preferential-rna-binding
#2
Jun Liu, Shilu Zhang, Chenghua Zhang, Jun Dong, Chengyi Shen, Jiang Zhu, Huajun Xu, Mingkai Fu, Guoqiang Yang, Xiaoming Zhang
By functionalizing triarylboron with cyclen, we developed a two-photon fluorescence probe, TAB-2, which can selectively bind RNA with a ratiometric readout. We tested TAB-2 in NIH/3T3 fibroblast cells, and demonstrated its capability in visualizing nucleoli and analyzing microenvironment polarity by two-photon and fluorescence-lifetime imaging microscopy.
October 6, 2017: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/28980425/investigating-the-healing-mechanisms-of-an-angiogenesis-promoting-topical-treatment-for-diabetic-wounds-using-multimodal-microscopy
#3
Joanne Li, Andrew J Bower, Zane Arp, Eric J Olson, Claire Holland, Eric J Chaney, Marina Marjanovic, Paritosh Pande, Aneesh Alex, Stephen A Boppart
Impaired skin wound healing is a significant co-morbid condition of diabetes that is caused by poor microcirculation, among other factors. Studies have shown that angiogenesis, a critical step in the wound healing process in diabetic wounds, can be promoted under hypoxia. In this study, an angiogenesis-promoting topical treatment for diabetic wounds, which promotes angiogenesis by mimicking a hypoxic environment via inhibition of prolyl hydroxylase resulting in elevation or maintenance of hypoxia inducible factor (HIF), was investigated utilizing a custom-built multimodal microscopy system equipped with phase-variance optical coherence tomography (PV-OCT) and fluorescence lifetime imaging microscopy (FLIM)...
October 5, 2017: Journal of Biophotonics
https://www.readbyqxmd.com/read/28936515/lysosomal-tracking-with-a-cationic-naphthalimide-using-multiphoton-fluorescence-lifetime-imaging-microscopy
#4
Meng Li, Haobo Ge, Vincenzo Mirabello, Rory L Arrowsmith, Gabriele Kociok-Köhn, Stanley W Botchway, Weihong Zhu, Sofia I Pascu, Tony D James
A naphthalimide-based chemosensing motif turns ON the fluorescence emission in solution in the presence of aqueous iron(iii) chloride, and maintains this property in living cancer cells. The emission response to Fe(iii) ions occurs simultaneously with a change in pH. The protonation of methyl piperazine-conjugated naphthalimide promotes its lysosomal localisation as assessed by co-localisation tests and fluorescence lifetime imaging microscopy (FLIM) in vitro.
September 22, 2017: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/28919329/phospholipase-c%C3%AE-interacts-with-cytosolic-partners-to-regulate-cell-proliferation
#5
REVIEW
Suzanne Scarlata, Ashima Singla, Osama Garwain
Phospholipase Cβ (PLCβ) is the main effector of the Gαq signaling pathway relaying different extracellular sensory information to generate intracellular calcium signals. Besides this classic function, we have found that PLCβ plays an important but unknown role in regulating PC12 cell differentiation by interacting with components in the RNA-induced silencing machinery. In trying to understand the role of PLCβ in PC12 cell differentiation, we find that over-expressing PLCβ reduces PC12 cell proliferation while down-regulating PLCβ increases the rate of cell proliferation...
September 9, 2017: Advances in Biological Regulation
https://www.readbyqxmd.com/read/28919280/soluble-gamma-secretase-modulators-attenuate-alzheimer-s-%C3%AE-amyloid-pathology-and-induce-conformational-changes-in-presenilin-1
#6
Frank Raven, Joseph F Ward, Katarzyna M Zoltowska, Yu Wan, Enjana Bylykbashi, Sean J Miller, Xunuo Shen, Se Hoon Choi, Kevin D Rynearson, Oksana Berezovska, Steven L Wagner, Rudolph E Tanzi, Can Zhang
A central pathogenic event of Alzheimer's disease (AD) is the accumulation of the Aβ42 peptide, which is generated from amyloid-β precursor protein (APP) via cleavages by β- and γ-secretase. We have developed a class of soluble 2-aminothiazole γ-secretase modulators (SGSMs) that preferentially decreases Aβ42 levels. However, the effects of SGSMs in AD animals and cells expressing familial AD mutations, as well as the mechanism of γ-secretase modulation remain largely unknown. Here, a representative of this SGSM scaffold, SGSM-36, was investigated using animals and cells expressing FAD mutations...
September 4, 2017: EBioMedicine
https://www.readbyqxmd.com/read/28874842/investigation-of-mitochondrial-metabolic-response-to-doxorubicin-in-prostate-cancer-cells-an-nadh-fad-and-tryptophan-flim-assay
#7
Shagufta Rehman Alam, Horst Wallrabe, Zdenek Svindrych, Ajay K Chaudhary, Kathryn G Christopher, Dhyan Chandra, Ammasi Periasamy
Prostate cancer (PCa) is one of the leading cancers in men in the USA. Lack of experimental tools that predict therapy response is one of the limitations of current therapeutic regimens. Mitochondrial dysfunctions including defective oxidative phosphorylation (OXPHOS) in cancer inhibit apoptosis by modulating ROS production and cellular signaling. Thus, correction of mitochondrial dysfunction and induction of apoptosis are promising strategies in cancer treatment. We have used Fluorescence Lifetime Imaging Microscopy (FLIM) to quantify mitochondrial metabolic response in PCa cells by tracking auto-fluorescent NAD(P)H, FAD and tryptophan (Trp) lifetimes and their enzyme-bound fractions as markers, before and after treatment with anti-cancer drug doxorubicin...
September 5, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28872817/carboplatin-loaded-raman-encoded-chitosan-coated-silver-nanotriangles-as-multimodal-traceable-nanotherapeutic-delivery-systems-and-ph-reporters-inside-human-ovarian-cancer-cells
#8
Monica Potara, Timea Nagy-Simon, Ana Maria Craciun, Sorina Suarasan, Emilia Licarete, Florica Imre-Lucaci, Simion Astilean
Ovarian cancer is a common cause of cancer death in women and is associated with the highest mortality rates of all gynecological malignancies. Carboplatin (CBP) is the most used cytotoxic agent in the treatment of ovarian cancer. Herein, we design and assess a CBP nanotherapeutic delivery system which allows combinatorial functionalities of chemotherapy, pH sensing, and multimodal traceable properties inside live NIH:OVCAR-3 ovarian cancer cells. In our design, a pH-sensitive Raman reporter, 4-mercaptobenzoic acid (4MBA) is anchored onto the surface of chitosan-coated silver nanotriangles (chit-AgNTs) to generate a robust surface-enhanced Raman scattering (SERS) traceable system...
September 27, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28863689/32-channel-time-correlated-single-photon-counting-system-for-high-throughput-lifetime-imaging
#9
P Peronio, I Labanca, G Acconcia, A Ruggeri, A A Lavdas, A A Hicks, P P Pramstaller, M Ghioni, I Rech
Time-Correlated Single Photon Counting (TCSPC) is a very efficient technique for measuring weak and fast optical signals, but it is mainly limited by the relatively "long" measurement time. Multichannel systems have been developed in recent years aiming to overcome this limitation by managing several detectors or TCSPC devices in parallel. Nevertheless, if we look at state-of-the-art systems, there is still a strong trade-off between the parallelism level and performance: the higher the number of channels, the poorer the performance...
August 2017: Review of Scientific Instruments
https://www.readbyqxmd.com/read/28862458/fluorescent-phase-pure-zero-dimensional-perovskite-related-cs4pbbr6-microdisks-synthesis-and-single-particle-imaging-study
#10
Sudipta Seth, Anunay Samanta
Quantum-confined perovskites are a new class of promising materials in optoelectronic applications. In this context, a zero-dimensional perovskite-related substance, Cs4PbBr6, having high exciton binding energy can be an important candidate, but its photoluminescence (PL) is a topic of recent debate. Herein, we report an ambient condition controlled synthesis of Cs4PbBr6 microdisks of different shapes and dimensions which exhibit fairly strong green PL (quantum yield up to 38%, band gap ∼2.43 eV) in the solid state...
September 6, 2017: Journal of Physical Chemistry Letters
https://www.readbyqxmd.com/read/28861826/in-vivo-interaction-studies-by-measuring-f%C3%A3-rster-resonance-energy-transfer-through-fluorescence-lifetime-imaging-microscopy-fret-flim
#11
Florian Fäßler, Peter Pimpl
Combinations of multiple fluorescent fusion proteins are commonly generated and used for colocalization studies in live cell imaging but also biochemical analysis of protein-protein interactions by co-immunoprecipitation in vitro. Advanced microscopy techniques like Förster resonance energy transfer through fluorescence lifetime imaging microscopy (FRET/FLIM) nowadays enable the combination of both approaches. This opens up the possibility to perform a location-specific protein-protein interaction analysis in vivo...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28837131/detection-of-nucleic-acid-protein-interactions-in-plant-leaves-using-fluorescence-lifetime-imaging-microscopy
#12
Laurent Camborde, Alain Jauneau, Christian Brière, Laurent Deslandes, Bernard Dumas, Elodie Gaulin
DNA-binding proteins (DNA-BPs) and RNA-binding proteins (RNA-BPs) have critical roles in living cells in all kingdoms of life. Various experimental approaches exist for the study of nucleic acid-protein interactions in vitro and in vivo, but the detection of such interactions at the subcellular level remains challenging. Here we describe how to detect nucleic acid-protein interactions in plant leaves by using a fluorescence resonance energy transfer (FRET) approach coupled to fluorescence lifetime imaging microscopy (FLIM)...
September 2017: Nature Protocols
https://www.readbyqxmd.com/read/28807980/cd177-modulates-human-neutrophil-migration-through-activation-mediated-integrin-and-chemoreceptor-regulation
#13
Ming Bai, Ricardo Grieshaber-Bouyer, Junxia Wang, Angela B Schmider, Zachary S Wilson, Liling Zeng, Olha Halyabar, Matthew D Godin, Hung N Nguyen, Anaïs Levescot, Pierre Cunin, Craig T Lefort, Roy J Soberman, Peter A Nigrovic
CD177 is a GPI-anchored protein expressed by a variable proportion of human neutrophils that mediates surface expression of the anti-neutrophil cytoplasmic antibody (ANCA) antigen proteinase 3. CD177 associates with β2 integrins and recognizes PECAM-1, suggesting a role in neutrophil migration. However, CD177(pos) neutrophils exhibit no clear migratory advantage in vivo, despite interruption of transendothelial migration by in vitro CD177 ligation. We sought to understand this paradox. Using a PECAM-1-independent transwell system, we found that CD177(pos) and CD177(neg) neutrophils migrated comparably...
August 14, 2017: Blood
https://www.readbyqxmd.com/read/28792278/astrocytic-endfoot-ca-2-correlates-with-parenchymal-vessel-responses-during-4-ap-induced-epilepsy-an-in%C3%A2-vivo-two-photon-lifetime-microscopy-study
#14
Cong Zhang, Maryam Tabatabaei, Samuel Bélanger, Hélène Girouard, Mohammad Moeini, Xuecong Lu, Frédéric Lesage
Neurovascular coupling (NVC) underlying the local increase in blood flow during neural activity forms the basis of functional brain imaging and is altered in epilepsy. Because astrocytic calcium (Ca(2+)) signaling is involved in NVC, this study investigates the role of this pathway in epilepsy. Here, we exploit 4-AP induced epileptic events to show that absolute Ca(2+) concentration in cortical astrocyte endfeet in vivo correlates with the diameter of precapillary arterioles during neural activity. We simultaneously monitored free Ca(2+) concentration in astrocytic endfeet with the Ca(2+)-sensitive indicator OGB-1 and diameter of adjacent arterioles in the somatosensory cortex of adult mice by two-photon fluorescence lifetime measurements following 4-AP injection...
January 1, 2017: Journal of Cerebral Blood Flow and Metabolism
https://www.readbyqxmd.com/read/28774048/tissue-like-phantoms-as-a-platform-for-inserted-fluorescence-nano-probes
#15
Tsviya Nayhoz, Eran A Barnoy, Dror Fixler
Tissue-like phantoms are widely used as a model for mimicking the optical properties of live tissue. This paper presents the results of a diffusion reflection method and fluorescence lifetime imaging microscopy measurements of fluorescein-conjugated gold nanorods in solution, as well as inserted in solid tissue-imitating phantoms. A lack of consistency between the fluorescence lifetime results of the solutions and the phantoms raises a question about the ability of tissue-like phantoms to maintain the optical properties of inserted contrast agents...
November 15, 2016: Materials
https://www.readbyqxmd.com/read/28763194/enhanced-visualization-of-hematoxylin-and-eosin-stained-pathological-characteristics-by-phasor-approach
#16
Teng Luo, Yuan Lu, Shaoxiong Liu, Danying Lin, Junle Qu
The phasor approach to fluorescence lifetime imaging microscopy (FLIM) is used to identify different types of tissues from hematoxylin and eosin (H&E) stained basal cell carcinoma (BCC) sections. The results suggest that working directly on the phasor space with the clustering assignment achieves immunofluorescence like simultaneous five or six-color imaging by using multiplexed fluorescence lifetimes of H&E. The phase approach is of particular effectiveness for enhanced visualization of the abnormal morphology of a suspected nidus...
September 5, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28754922/shadowy-a-dark-yellow-fluorescent-protein-for-flim-based-fret-measurement
#17
Hideji Murakoshi, Akihiro C E Shibata
Fluorescence lifetime imaging microscopy (FLIM)-based Förster resonance energy transfer (FRET) measurement (FLIM-FRET) is one of the powerful methods for imaging of intracellular protein activities such as protein-protein interactions and conformational changes. Here, using saturation mutagenesis, we developed a dark yellow fluorescent protein named ShadowY that can serve as an acceptor for FLIM-FRET. ShadowY is spectrally similar to the previously reported dark YFP but has a much smaller quantum yield, greater extinction coefficient, and superior folding property...
July 28, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28749340/super-resolution-imaging-of-synaptic-and-extra-synaptic-ampa-receptors-with-different-sized-fluorescent-probes
#18
Sang Hak Lee, Chaoyi Jin, En Cai, Pinghua Ge, Yuji Ishitsuka, Kai Wen Teng, Andre A de Thomaz, Duncan Nall, Murat Baday, Okunola Jeyifous, Daniel Demonte, Christopher M Dundas, Sheldon Park, Jary Y Delgado, William N Green, Paul R Selvin
Previous studies tracking AMPA receptor (AMPAR) diffusion at synapses observed a large mobile extrasynaptic AMPAR pool. Using super-resolution microscopy, we examined how fluorophore size and photostability affected AMPAR trafficking outside of, and within, post-synaptic densities (PSDs) from rats. Organic fluorescent dyes (≈4 nm), quantum dots, either small (≈10 nm diameter; sQDs) or big (>20 nm; bQDs), were coupled to AMPARs via different-sized linkers. We find that >90% of AMPARs labeled with fluorescent dyes or sQDs were diffusing in confined nanodomains in PSDs, which were stable for 15 min or longer...
July 27, 2017: ELife
https://www.readbyqxmd.com/read/28723155/inhibiting-the-fibrillation-of-serum-albumin-proteins-in-the-presence-of-surface-active-ionic-liquids-sails-at-low-ph-spectroscopic-and-microscopic-study
#19
Sangita Kundu, Chiranjib Banerjee, Nilmoni Sarkar
One of the key necessary steps to prevent human neurological disorders is the efficient disruption of protein aggregation or amyloid fibril. In this article, we have explored the effect of three amphiphilic surface active ionic liquids (SAILs), namely 1-methyl-3-octylimidazolium chloride ([C8mim]Cl), 1-dodecyl-3-methyllimidazolium chloride ([C12mim]Cl), and 1-hexadecyl-3-methyllimidazolium chloride ([C16mim]Cl) having concentrations of 5.8, 0.29, and 0.08 mM, respectively, on bovine serum albumin (BSA) and human serum albumin (HSA) fibril...
August 4, 2017: Journal of Physical Chemistry. B
https://www.readbyqxmd.com/read/28718466/exploring-viscosity-polarity-and-temperature-sensitivity-of-bodipy-based-molecular-rotors
#20
Aurimas Vyšniauskas, Ismael López-Duarte, Nicolas Duchemin, Thanh-Truc Vu, Yilei Wu, Ekaterina M Budynina, Yulia A Volkova, Eduardo Peña Cabrera, Diana E Ramírez-Ornelas, Marina K Kuimova
Microviscosity is a key parameter controlling the rate of diffusion and reactions on the microscale. One of the most convenient tools for measuring microviscosity is by fluorescent viscosity sensors termed 'molecular rotors'. BODIPY-based molecular rotors in particular proved extremely useful in combination with fluorescence lifetime imaging microscopy, for providing quantitative viscosity maps of living cells as well as measuring dynamic changes in viscosity over time. In this work, we investigate several new BODIPY-based molecular rotors with the aim of improving on the current viscosity sensing capabilities and understanding how the structure of the fluorophore is related to its function...
September 27, 2017: Physical Chemistry Chemical Physics: PCCP
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