Intracellular temperature mapping with a fluorescent polymeric thermometer and fluorescence lifetime imaging microscopy

Okabe, Kimiko; Inada, Noriko; Gota, Chie; Harada, Yoshie; Funatsu, Takashi; Uchiyama, Seiichi

doi:10.1038/ncomms1714

Cited by 1,009 publications

(926 citation statements)

References 45 publications

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“…Another perspective is the development of OCP-based biosensors, since the obtained fluorescent OCP-TMR complexes could be used as novel molecular sensors of light intensity, temperature, and viscosity, which would obviate the need for complex instrumentation, such as fluorescence correlation spectroscopy, to measure these parameters. The first intracellular temperature mapping methods based on fluorescent thermometers and fluorescence-lifetime imaging microscopy were developed recently (43)(44)(45). The observed dependence of the OCP conversion rate on temperature and viscosity is consistent with the general behavior expected for protein folding: whereas temperature dependence followed the Arrhenius law, the dependence on viscosity was in line with the Kramers theory.…”

Section: Eetsupporting

confidence: 68%

Fluorescent Labeling Preserving OCP Photoactivity Reveals Its Reorganization during the Photocycle

Maksimov

Sluchanko

Mironov

et al. 2017

Biophysical Journal

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Orange carotenoid protein (OCP), responsible for the photoprotection of the cyanobacterial photosynthetic apparatus under excessive light conditions, undergoes significant rearrangements upon photoconversion and transits from the stable orange to the signaling red state. This is thought to involve a 12-Å translocation of the carotenoid cofactor and separation of the N-and C-terminal protein domains. Despite clear recent progress, the detailed mechanism of the OCP photoconversion and associated photoprotection remains elusive. Here, we labeled the OCP of Synechocystis with tetramethylrhodamine-maleimide (TMR) and obtained a photoactive OCP-TMR complex, the fluorescence of which was highly sensitive to the protein state, showing unprecedented contrast between the orange and red states and reflecting changes in protein conformation and the distances from TMR to the carotenoid throughout the photocycle. The OCP-TMR complex was sensitive to the light intensity, temperature, and viscosity of the solvent. Based on the observed Fö rster resonance energy transfer, we determined that upon photoconversion, the distance between TMR (donor) bound to a cysteine in the C-terminal domain and the carotenoid (acceptor) increased by 18 Å , with simultaneous translocation of the carotenoid into the N-terminal domain. Time-resolved fluorescence anisotropy revealed a significant decrease of the OCP rotation rate in the red state, indicating that the light-triggered conversion of the protein is accompanied by an increase of its hydrodynamic radius. Thus, our results support the idea of significant structural rearrangements of OCP, providing, to our knowledge, new insights into the structural rearrangements of OCP throughout the photocycle and a completely novel approach to the study of its photocycle and non-photochemical quenching. We suggest that this approach can be generally applied to other photoactive proteins.

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Section: Eetsupporting

confidence: 68%

Fluorescent Labeling Preserving OCP Photoactivity Reveals Its Reorganization during the Photocycle

Maksimov

Sluchanko

Mironov

et al. 2017

Biophysical Journal

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“…When combined with precision temperature measurement it promises to give excess to small temperature changes caused e.g. by chemical reactions or biochemical processes [2]. However, nanoscale temperature measurements and precision have excluded each other so far owing to the physical processes used for temperature measurement of limited stability of nanoscale probes [3].…”

mentioning

confidence: 99%

High-Precision Nanoscale Temperature Sensing Using Single Defects in Diamond

et al. 2013

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Measuring local temperature with a spatial resolution on the order of a few nanometers has a wide range of applications from semiconductor industry over material to life sciences [1]. When combined with precision temperature measurement it promises to give excess to small temperature changes caused e.g. by chemical reactions or biochemical processes [2]. However, nanoscale temperature measurements and precision have excluded each other so far owing to the physical processes used for temperature measurement of limited stability of nanoscale probes [3]. Here we experimentally demonstrate a novel nanoscale temperature sensing technique based on single atomic defects in diamonds. Sensor sizes range from millimeter down to a few tens of nanometers. Utilizing the sensitivity of the optically accessible electron spin level structure to temperature changes [4] we achieve a temperature noise floor of 5 mK/ √ Hz for single defects in bulk sensors. Using doped nanodiamonds as sensors yields temperature measurement with 130 mK/ √ Hz noise floor and accuracies down to 1 mK at length scales of a few ten nanometers. The high sensitivity to temperature changes together with excellent spatial resolution combined with outstanding sensor stability allows for nanoscale precision temperature determination enough to measure chemical processes of few or single molecules by their reaction heat even in heterogeneous environments like cells.Several kinds of nanoscale temperature sensing techniques have been developed in the recent past [1]. These are scanning thermal microscopes (SThM) [5], dispersed or scanned individual nanoprobes [3,6], direct methods like micro-Raman spectroscopy [7] or near-field optical temperature measurements [8]. SThMs have temperature sensitive elements at a scanning tip (e.g. thermocouple), the nanoprobes have temperature dependent properties (e.g. fluorescence spectrum) which can be accessed without direct contact.In this study utilize a single quantum system in a solid state matrix as a temperature nanoprobe, namely the negatively charged nitrogen-vacancy (NV) center in diamond, which allows probe sizes down to ∼ 5 nm [9]. High fidelity control of its ground state electronic and nuclear spins has been demonstrated for various quantum information test experiments [10-15] as well as for nanometer scale metrology purposes [16][17][18][19] e.g. measuring small magnetic and electric fields. Here we show that it also allows tracking temperature with high precision. Temperature nanoprobes can be either dispersed in the specimen to be investigated or used in scanning probe geometry (see fig. 1a).The NV center is a molecular impurity in diamond comprising a substitutional nitrogen impurity and an adjacent carbon vacancy. Optical excitation in a wavelength range from 460 nm to 580 nm yields intense fluorescence emission [20]. Excitation also leads to a high degree of ground state electron spin polarization (S = 1, the actual sensor level) into its m S = 0 (|0 ) sublevel [21]. Furthermore the fluorescence decreases upo...

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“…For instance, in vivo observation of temperature in tissue using PL material has absorbed much attentions recently, 10) and is considered to be helpful for a medical treatment, in which temperature-monitoring is a key to improving the quality of the treatment. 11) Zirconia (ZrO 2 ) is a wide-bandgap dielectrics, and is also utilized to synthesize an engineering ceramics and artificial teeth because of its high chemical durability and harmless.…”

Section: )7)mentioning

confidence: 99%

Formation and photoluminescence of zirconia dendrites in borosilicate glass-ceramics

Nobuta

Takahashi

Miyazaki

et al. 2017

J. Ceram. Soc. Japan

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We prepared a borosilicate glass, in which the ZrO 2 phase is singly crystallizable, i.e., 15Na 2 O15ZrO 2 30B 2 O 3 40SiO 2 glass, and investigated the texture and morphology of the resulting glass-ceramics. ZrO 2 dendrites with a tetragonal system (hightemperature phase) were developed as initial phase, and the tetragonal phase was transformed to the monoclinic phase (lowtemperature phase) by elongation of the heat-treatment time, and finally the needle-/rod-like crystals on a scale of few hundred nanometers were obtained. The glass-ceramics with monoclinic ZrO 2 showed photoluminescence around 480 nm by excitation in the ultraviolet region. The effect of TiO 2 -addition on the photoluminescent property was also considered.

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Intracellular temperature mapping with a fluorescent polymeric thermometer and fluorescence lifetime imaging microscopy

Cited by 1,009 publications

References 45 publications

Fluorescent Labeling Preserving OCP Photoactivity Reveals Its Reorganization during the Photocycle

Fluorescent Labeling Preserving OCP Photoactivity Reveals Its Reorganization during the Photocycle

High-Precision Nanoscale Temperature Sensing Using Single Defects in Diamond

Formation and photoluminescence of zirconia dendrites in borosilicate glass-ceramics

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