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Near Infrared FLIM

Tsukasa Funane, Steven S Hou, Katarzyna Marta Zoltowska, Susanne J van Veluw, Oksana Berezovska, Anand T N Kumar, Brian J Bacskai
We have developed an imaging technique which combines selective plane illumination microscopy with time-domain fluorescence lifetime imaging microscopy (SPIM-FLIM) for three-dimensional volumetric imaging of cleared mouse brains with micro- to mesoscopic resolution. The main features of the microscope include a wavelength-adjustable pulsed laser source (Ti:sapphire) (near-infrared) laser, a BiBO frequency-doubling photonic crystal, a liquid chamber, an electrically focus-tunable lens, a cuvette based sample holder, and an air (dry) objective lens...
May 2018: Review of Scientific Instruments
Yingying Ning, Juan Tang, Yi-Wei Liu, Jing Jing, Yuansheng Sun, Jun-Long Zhang
Herein, we report the design and synthesis of biocompatible Yb3+ complexes for near-infrared (NIR) living cell imaging. Upon excitation at either the visible (Soret band) or red region (Q band), these β-fluorinated Yb3+ complexes display high NIR luminescence (quantum yields up to 23% and 13% in dimethyl sulfoxide and water, respectively) and have higher stabilities and prolonged decay lifetimes (up to 249 μs) compared to the β-non-fluorinated counterparts. This renders the β-fluorinated Yb3+ complexes as a new class of biological optical probes in both steady-state imaging and time-resolved fluorescence lifetime imaging (FLIM)...
April 21, 2018: Chemical Science
Sorina Suarasan, Emilia Licarete, Simion Astilean, Ana-Maria Craciun
Nowadays, the non-linear optical effect of two-photon excited (TPE) fluorescence has recently grown in interest in recent years over other optical imaging method, due to improved 3D spatial resolution, deep penetrability and less photodamage of living organism owing to the excitation in near-infrared region (NIR). In parallel, gold nanoparticles (AuNPs) have gain considerable attention for NIR TPE bio-imaging applications due to their appealing ability to generate strong intrinsic photoluminescence (PL). Here, we demonstrate the capability of differently shaped gelatin-coated AuNPs to perform as reliable label-free contrast agents for the non-invasive NIR imaging of NIH:OVCAR-3 ovary cancer cells via TPE Fluorescence Lifetime Imaging Microscopy (FLIM)...
June 1, 2018: Colloids and Surfaces. B, Biointerfaces
Annette Buntz, Sarah Wallrodt, Eva Gwosch, Michael Schmalz, Sascha Beneke, Elisa Ferrando-May, Andreas Marx, Andreas Zumbusch
Poly(ADP-ribos)ylation (PARylation) is an important posttranslational protein modification, and is involved in major cellular processes such as gene regulation and DNA repair. Its dysregulation has been linked to several diseases, including cancer. Despite its importance, methods to observe PARylation dynamics within cells are rare. By following a chemical biology approach, we developed a fluorescent NAD(+) analogue that proved to be a competitive building block for protein PARylation in vitro and in cells...
September 5, 2016: Angewandte Chemie
N Rubio, L M Hirvonen, E Z Chong, J T W Wang, M Bourgognon, H Kafa, H A F M Hassan, W T Al-Jamal, D McCarthy, C Hogstrand, F Festy, K T Al-Jamal
The intrinsic nonlinear photoluminescence (PL) property of chemically functionalized multi-walled nanotubes MWNTs (f-MWNTs) is reported in this study. f-MWNTs are imaged in fixed lung epithelial cancer cells (A549) and Kupffer cells in vitro, and in subcutaneously implanted solid tumors in vivo, for the first time, using multiphoton PL and fluorescence lifetime imaging (FLIM). Multiphoton imaging in the near-infrared excitation region (∼750-950 nm), employed in this study in a label-free manner, provides sensitivity and resolution optimal to track f-MWNTs within intra-cellular compartments and facilitates tumour imaging and sentinel lymph node tracking in vivo...
June 7, 2015: Chemical Communications: Chem Comm
Stanley W Botchway, Kathrin M Scherer, Steve Hook, Christopher D Stubbs, Eleanor Weston, Roger H Bisby, Anthony W Parker
Multiphoton microscopy is widely employed in the life sciences using extrinsic fluorescence of low- and high-molecular weight labels with excitation and emission spectra in the visible and near infrared regions. For imaging of intrinsic and extrinsic fluorophores with excitation spectra in the ultraviolet region, multiphoton excitation with one- or two-colour lasers avoids the need for ultraviolet-transmitting excitation optics and has advantages in terms of optical penetration in the sample and reduced phototoxicity...
April 2015: Journal of Microscopy
Yongkui Xu, Ruoyu He, Dongdong Lin, Minbiao Ji, Jiyao Chen
A new method to image drug release from drug-nanoparticle composites in living cells was established. The composites of silica coated gold nanorods (AuNR@SiO2) and chlorine e6 (Ce6) photosensitizers (AuNR@SiO2-Ce6) were formed by electrostatic force with a Ce6 loading efficiency of 80%. The strong resonance absorptions of AuNR@SiO2-Ce6 in the near-infrared (NIR) region enabled the effective release of Ce6 from AuNR@SiO2-Ce6 by 780 nm CW laser irradiation. The 780 nm laser beam was applied to not only control the releasing amount of Ce6 from cellular AuNR@SiO2-Ce6 by adjusting the irradiation dose (time), but also to spatially confine the Ce6 release in cells by focusing the laser beam on the target sites...
February 14, 2015: Nanoscale
Himanshu Sharma, Michelle A Digman, Natasha Felsinger, Enrico Gratton, Michelle Khine
We introduce a manufacturable and scalable method for creating tunable wrinkled ferromagnetic-metallic structures to enhance fluorescence signals. Thin layers of nickel (Ni) and gold (Au) were deposited onto a pre-stressed thermoplastic (shrink wrap film) polymer. Heating briefly forced the metal films to buckle when the thermoplastic retracted, resulting in multi-scale composite 'wrinkles'. This is the first demonstration of leveraging the plasmons in such hybrid nanostructures by metal enhanced fluorescence (MEF) in the near-infrared wavelengths...
2014: Optical Materials Express
R Nothdurft, P Sarder, S Bloch, J Culver, S Achilefu
Although single-photon fluorescence lifetime imaging microscopy (FLIM) is widely used to image molecular processes using a wide range of excitation wavelengths, the captured emission of this technique is confined to the visible spectrum. Here, we explore the feasibility of utilizing near-infrared (NIR) fluorescent molecular probes with emission >700 nm for FLIM of live cells. The confocal microscope is equipped with a 785 nm laser diode, a red-enhanced photomultiplier tube, and a time-correlated single photon counting card...
August 2012: Journal of Microscopy
Li Shang, Naghmeh Azadfar, Florian Stockmar, Winfried Send, Vanessa Trouillet, Michael Bruns, Dagmar Gerthsen, G Ulrich Nienhaus
A facile strategy to synthesize water-soluble fluorescent gold nanoclusters (Au NCs) stabilized with the bidentate ligand dihydrolipoic acid (DHLA) is reported. The DHLA-capped Au NCs are characterized by UV-vis absorption spectroscopy, fluorescence spectroscopy, transmission electron microscopy, and X-ray photoelectron spectroscopy. The Au NCs possess many attractive features including ultrasmall size, bright near-infrared luminescence, high colloidal stability, and good biocompatibility, making them promising imaging agents for biomedical and cellular imaging applications...
September 19, 2011: Small
Rafal Luchowski, Mariusz Szabelski, Pabak Sarkar, Elisa Apicella, Krishna Midde, Sangram Raut, Julian Borejdo, Zygmunt Gryczynski, Ignacy Gryczynski
We studied the fluorescence properties of several potential picosecond lifetime standards suitable for two-photon excitation from a Ti:sapphire femtosecond laser. The fluorescence emission of the selected fluorophores (rose bengal, pyridine 1, and LDS 798) covered the visible to near-infrared wavelength range from 550 to 850 nm. We suggest that these compounds can be used to measure the appropriate instrument response functions needed for accurate deconvolution of fluorescence lifetime data. Lifetime measurements with multiphoton excitation that use scatterers as a reference may fail to properly resolve fluorescence intensity decays...
August 2010: Applied Spectroscopy
Johannes T Wessels, Kensuke Yamauchi, Robert M Hoffman, Fred S Wouters
This review focuses on technical advances in fluorescence microscopy techniques including laser scanning techniques, fluorescence-resonance energy transfer (FRET) microscopy, fluorescence lifetime imaging (FLIM), stimulated emission depletion (STED)-based super-resolution microscopy, scanning confocal endomicroscopes, thin-sheet laser imaging microscopy (TSLIM), and tomographic techniques such as early photon tomography (EPT) as well as on clinical laser-based endoscopic and microscopic techniques. We will also discuss the new developments in the field of fluorescent dyes and fluorescent genetic reporters that enable new possibilities in high-resolution and molecular imaging both in in vitro and in vivo...
July 2010: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
Keiichi Sugata, Shingo Sakai, Nakagawa Noriaki, Osamu Osanai, Takashi Kitahara, Yoshinori Takema
BACKGROUND/PURPOSE: Multiphoton fluorescence lifetime imaging (FLIM) is a technique that produces an image based on differences in the decay rate of fluorescence from a sample. Based on this method, the DermaInspect was developed to observe human skin components non-invasively. In this study, we used the DermaInspect to study melanin in skin. METHODS: A human three-dimensional skin model containing melanocytes was embedded in an OCT compound, frozen and sectioned at 10 microm...
February 2010: Skin Research and Technology
Bryan J McCranor, Richard B Thompson
With the increased development and use of fluorescence lifetime-based sensors, fiber optic sensors, fluorescence lifetime imaging microscopy (FLIM), and plate and array readers, , calibration standards are essential to ensure the proper function of these devices and accurate results. For many devices that utilize a "front face excitation" geometry where the excitation is nearly coaxial with the direction of emission, scattering-based lifetime standards are problematic and fluorescent lifetime standards are necessary...
March 2010: Journal of Fluorescence
Sylvain Gioux, Vida Kianzad, Razvan Ciocan, Hak Soo Choi, Chad Nelson, Jeffrey Thumm, Robert J Filkins, Stephen J Lomnes, John V Frangioni
Near-infrared (NIR) fluorescence has the potential to provide surgeons with real-time intraoperative image-guidance. Increasing the signal-to-background ratio of fluorescent agents involves delivering a controllable excitation fluence rate of proper wavelength and/or using complementary imaging techniques such as FLIM. In this study we describe a low-cost linear driver circuit capable of driving Light Emitting Diodes (LEDs) from DC to 35 MHz, at high power, and which permit fluorescence CW and lifetime measurements...
2008: Proceedings of SPIE
Daniel Doerr, Martin Stark, Friederike Ehrhart, Heiko Zimmermann, Frank Stracke
In this study we demonstrate a new noninvasive imaging method to monitor freezing processes in biological samples and to investigate life in the frozen state. It combines a laser scanning microscope with a computer-controlled cryostage. Nearinfrared (NIR) femtosecond laser pulses evoke the fluorescence of endogenous fluorophores and fluorescent labels due to multiphoton absorption.The inherent optical nonlinearity of multiphoton absorption allows 3D fluorescence imaging for optical tomography of frozen biological material in-situ...
August 2009: Biotechnology Journal
E Werkmeister, H Kerdjoudj, L Marchal, J F Stoltz, D Dumas
Imaging thick and opaque tissue, like blood vessel, in a noninvasive mode with high resolution, is nowadays possible with multiphoton technology. A near-infrared excitation presents the advantage to be compatible with living specimens and allows a deep penetration into tissues. The nonlinear excitation process is followed by several deactivation ways, among which fluorescence emission can be represented with Spectral or Lifetime imaging. Applied to ex vivo blood vessel imaging, these techniques enabled us to discriminate cell structures (nucleus, cytoskeleton) by fluorescent labelling (Hoechst, QDots)...
2007: Clinical Hemorheology and Microcirculation
Alexander Ehlers, Iris Riemann, Martin Stark, Karsten König
In vivo and in vitro multiphoton imaging was used to perform high resolution optical sectioning of human hair by nonlinear excitation of endogenous as well as exogenous fluorophores. Multiphoton fluorescence lifetime imaging (FLIM) based on time-resolved single photon counting and near-infrared femtosecond laser pulse excitation was employed to analyze the various fluorescent hair components. Time-resolved multiphoton imaging of intratissue pigments has the potential (i) to identify endogenous keratin and melanin, (ii) to obtain information on intrahair dye accumulation, (iii) to study bleaching effects, and (iv) to monitor the intratissue diffusion of pharmaceutical and cosmetical components along hair shafts...
February 2007: Microscopy Research and Technique
Anuradha Godavarty, Eva M Sevick-Muraca, Margaret J Eppstein
Near-infrared fluorescence tomography using molecularly targeted lifetime-sensitive, fluorescent contrast agents have applications for early-stage cancer diagnostics. Yet, although the measurement of fluorescent lifetime imaging microscopy (FLIM) is extensively used in microscopy and spectroscopy applications, demonstration of fluorescence lifetime tomography for medical imaging is limited to two-dimensional studies. Herein, the feasibility of three-dimensional fluorescence-lifetime tomography on clinically relevant phantom volumes is established, using (i) a gainmodulated intensified charge coupled device (CCD) and modulated laser diode imaging system, (ii) two fluorescent contrast agents, e...
April 2005: Medical Physics
Volker Ulrich, Peter Fischer, Iris Riemann, Karsten Königt
An inverted fluorescence microscope was upgraded into a compact three-dimensional laser scanning microscope (LSM) of 65 x 62 x 48 cm dimensions by means of a fast kHz galvoscanner unit, a piezodriven z-stage, and a picosecond (ps) 50 MHz laser diode at 405 nm. In addition, compact turn-key near infrared femtosecond lasers have been employed to perform multiphoton fluorescence and second harmonic generation (SHG) microscopy. To expand the features of the compact LSM, a time-correlated single photon counting unit as well as a Sagnac interferometer have been added to realize fluorescence lifetime imaging (FLIM) and spectral imaging...
September 2004: Scanning
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