Studying Membrane Properties Using Fluorescence Lifetime Imaging Microscopy (FLIM)

Stöckl, Martin; Bizzarri, Ranieri; Subramaniam, Vinod

doi:10.1007/4243_2012_48

Cited by 6 publications

(3 citation statements)

References 97 publications

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“…FLIM allows the direct visualization of lipid domains and interactions of membrane proteins with each other or with lipid domains by fluorescence lifetime scanning microscopy [30], as well as the fluorescence imaging of cancerous tissues in live organisms [31]. Moreover, the current generation of modern confocal instruments produces imaging scans acquired with very high resolution, also known as super-resolution images [32,33].…”

Section: Fluorescence Lifetime Imagingmentioning

confidence: 99%

Using fluorescence for studies of biological membranes: a review

Kyrychenko¹

2015

Methods Appl. Fluoresc.

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Fluorescence techniques have become powerful and widely used tools for studies of biochemical and biophysical processes occurring in biological membranes. Various fluorescence methods have played and continue to play key roles in modern membrane science, so that there have been several focused reviews on this topic. Here, I present the progress and recent achievements in various fluorescence approaches commonly utilized in studies of biological membranes. Applications of numerous fluorescence methods have been reviewed, including single molecule detection, confocal scanning fluorescence microscopy and fluorescence lifetime imaging. I focus on the benefits and limitations of various fluorescence techniques and their combinations, as well as the available methods of in vivo studying. A separate section is dedicated to discussing and comparing different classes of fluorescent membrane probes and their applications to the study of biological membranes. The review should provide researchers from chemistry, biochemistry, and biophysics with the necessary background to identify a range of suitable fluorescence methods in order to successfully design and conduct experimental studies on model lipid bilayers and biological membranes.

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Section: Fluorescence Lifetime Imagingmentioning

confidence: 99%

Using fluorescence for studies of biological membranes: a review

Kyrychenko¹

2015

Methods Appl. Fluoresc.

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“…In particular, fluorescence measurements can provide information not only on the specific molecular makeup of a sample, but also on the local environment surrounding the fluorescence molecule or fluorophore (such as pH, ion concentrations, etc. ), which give value to the fluorescence based techniques a powerful analysis tool [3,4,5,6]. However, the fluorescence techniques based on intensity measurements are prone to misinterpretation due to their dependence on parameters such as excitation light intensity and fluorophore concentration.…”

Section: Introductionmentioning

confidence: 99%

A Point-of-Care Device for Molecular Diagnosis Based on CMOS SPAD Detectors with Integrated Microfluidics

Canals¹,

Franch²,

Alonso³

et al. 2019

Sensors

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We describe the integration of techniques and technologies to develop a Point-of-Care for molecular diagnosis PoC-MD, based on a fluorescence lifetime measurement. Our PoC-MD is a low-cost, simple, fast, and easy-to-use general-purpose platform, aimed at carrying out fast diagnostics test through label detection of a variety of biomarkers. It is based on a 1-D array of 10 ultra-sensitive Single-Photon Avalanche Diode (SPAD) detectors made in a 0.18 μm High-Voltage Complementary Metal Oxide Semiconductor (HV-CMOS) technology. A custom microfluidic polydimethylsiloxane cartridge to insert the sample is straightforwardly positioned on top of the SPAD array without any alignment procedure with the SPAD array. Moreover, the proximity between the sample and the gate-operated SPAD sensor makes unnecessary any lens or optical filters to detect the fluorescence for long lifetime fluorescent dyes, such as quantum dots. Additionally, the use of a low-cost laser diode as pulsed excitation source and a Field-Programmable Gate Array (FPGA) to implement the control and processing electronics, makes the device flexible and easy to adapt to the target label molecule by only changing the laser diode. Using this device, reliable and sensitive real-time proof-of-concept fluorescence lifetime measurement of quantum dot QdotTM 605 streptavidin conjugate is demonstrated.

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“…Thus, slowing the twisting motions as viscosity increases leads to stronger emission and longer lifetime decay. 8 The ForsterHoffmann model of molecular rotors 9 affords a reliable quantitative description of fluorescence dependence on viscosity for most FMRs. 10 Nonetheless, a common drawback of FMRs is the difficulty to predict their sensitivity towards local polarity, owing to the complex dipolar properties of accessible excited states.…”

Section: Introductionmentioning

confidence: 99%

A fluorescent molecular rotor showing vapochromism, aggregation-induced emission, and environmental sensing in living cells

et al. 2016

Self Cite

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Among the plethora of recently proposed molecular sensors, those belonging to the class of fluorescent molecular rotors (FMRs) have attracted much attention owing to their peculiar photophysical properties that enable an unprecedented sensitivity towards environmental microviscosity. The usual FMR synthetic design prescribes chromophores characterized by an intramolecular rotation between two well-defined excited states, a locally excited state and a twisted internal charge transfer (TICT) state, where the sensing capabilities arise from a dual competition of the corresponding radiative/non-radiative decay processes. However, we have recently demonstrated a different modus operandi of a new subclass of solvatochromic FMRs, which exploit a solvent-independent, barrier-free intramolecular rotation of the excited dye. The rotational dynamics is modulated by local viscosity and, in turn, manifested through a variable spectral signal. In order to translate the same rotational mechanism in a versatile sensor of polarity and viscosity, we designed and thoroughly characterized a novel FMR, namely 4-(triphenylamino)-phthalonitrile (TPAP). Remarkably, in addition to a high sensitivity versus solvent polarity and viscosity, TPAP is also able to form stable fluorescent nanoparticles characterized by aggregation-induced emission, via a simple sonochemical treatment. Such peculiar features are tested in different applications aiming at illustrating its capability to report on solvatochromic and vapochromic effects, as well as to provide detailed intracellular information through bioimaging studies.

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Studying Membrane Properties Using Fluorescence Lifetime Imaging Microscopy (FLIM)

Cited by 6 publications

References 97 publications

Using fluorescence for studies of biological membranes: a review

Using fluorescence for studies of biological membranes: a review

A Point-of-Care Device for Molecular Diagnosis Based on CMOS SPAD Detectors with Integrated Microfluidics

A fluorescent molecular rotor showing vapochromism, aggregation-induced emission, and environmental sensing in living cells

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