Fluorescence Thermometer Research Articles

Temperature imaging using a cationic linear fluorescent polymeric thermometer and fluorescence lifetime imaging microscopy.

Temperature is one of the most important of the physiological parameters that determine the biological status of living organisms. However, intracellular temperature was not imaged at the single-cell level until recently because of the lack of a molecular thermometer that can be applied to living cells. We have recently developed a method for imaging intracellular temperature using a cationic linear fluorescent polymeric thermometer (FPT) and fluorescence lifetime imaging microscopy (FLIM). The cationic linear FPT exhibits cell permeability in various mammalian cell lines and yeast cells, entering live cells within 10 min of incubation. Intracellular thermometry using the cationic linear FPT and FLIM can be used to image temperature with high temperature resolution (0.3-1.29 °C within a temperature range of 25-35 °C). The diffuse intracellular localization of the cationic linear FPT allows a high spatial resolution (i.e., the light microscope's diffraction limit, 200 nm), enabling the detection of temperature distributions at the subcellular level. This protocol, including the construction of a calibration curve and intracellular temperature imaging, requires ~14 h. Experience in handling cultured mammalian cells and use of a confocal laser-scanning microscope (CLSM) is required.

2,2'-Diamino-6,6'-diboryl-1,1'-binaphthyl: A Versatile Building Block for Temperature-Dependent Dual Fluorescence and Switchable Circularly Polarized Luminescence.

Temperature-dependent dual fluorescence and switchable circularly polarized luminescence (CPL) are two highly pursued but challenging properties for small organic molecules (SOMs). We herein disclose a triarylborane π-system based on a 2,2'-diamino-6,6'-diboryl-1,1'-binaphthyl scaffold that can serve as a versatile building block for achieving these two properties by simply choosing different amino groups. BNMe2 -BNaph with less bulky dimethylamino groups displays temperature-dependent dual fluorescence, and can thus be used as a highly sensitive ratiometric fluorescence thermometer. On the other hand, BNPh2 -BNaph with bulky diphenylamino groups exhibits intense fluorescence in both solution and in the solid state. A change of solvent from nonpolar cyclohexane to highly polar MeCN not only shifts the CPL position to much longer wavelength but also inverts the CPL sign. In addition, the complexation of BNPh2 -BNaph with fluoride greatly enhances the CPL intensity.

2,2′‐Diamino‐6,6′‐diboryl‐1,1′‐binaphthyl: A Versatile Building Block for Temperature‐Dependent Dual Fluorescence and Switchable Circularly Polarized Luminescence

AbstractTemperature‐dependent dual fluorescence and switchable circularly polarized luminescence (CPL) are two highly pursued but challenging properties for small organic molecules (SOMs). We herein disclose a triarylborane π‐system based on a 2,2′‐diamino‐6,6′‐diboryl‐1,1′‐binaphthyl scaffold that can serve as a versatile building block for achieving these two properties by simply choosing different amino groups. BNMe2‐BNaph with less bulky dimethylamino groups displays temperature‐dependent dual fluorescence, and can thus be used as a highly sensitive ratiometric fluorescence thermometer. On the other hand, BNPh2‐BNaph with bulky diphenylamino groups exhibits intense fluorescence in both solution and in the solid state. A change of solvent from nonpolar cyclohexane to highly polar MeCN not only shifts the CPL position to much longer wavelength but also inverts the CPL sign. In addition, the complexation of BNPh2‐BNaph with fluoride greatly enhances the CPL intensity.

A ratiometric fluorescent thermometer based on amphiphilic alkynylpyrene derivatives

A new alkynylpyrene derivative shows thermoresponsive, ratiometric, and reversible fluorescence color switching from red to green in an ambient atmosphere.

Fluorescent thermometer based on a quinolinemalononitrile copolymer with aggregation-induced emission characteristics

An AIEgen-grafted copolymer P(NIPAM-co-EM) is successfully constructed with thermo-responsive LCST-featuring matrix PNIPAM for temperature detection.

Fluorescent In based MOFs showing “turn on” luminescence towards thiols and acting as a ratiometric fluorescence thermometer

A chiral 2D In-MOF shows unique “turn on” luminescence towards thiols and acts as a ratiometric fluorescent sensor for temperature upon incorporation of Eu3+.

An ultrasensitive ratiometric fluorescent thermometer based on frustrated static excimers in the physiological temperature range.

A ratiometric fluorescent thermometer (RFT) based on the frustrated static excimers (FSEs) of DEH-PDI (N,N'-di(2-ethylhexyl)-3,4,9,10-perylenetetracarboxylic diimide) was designed and synthesized. The RFT exhibited ultrahigh sensitivities, which far exceed the existing RFTs in the physiological temperature range. The RFT also showed outstanding precisions, stability, and an obvious thermochromism in the temperature range.

Fixable Molecular Thermometer for Real-Time Visualization and Quantification of Mitochondrial Temperature.

A change of mitochondrial temperature can be an important indicator of mitochondrial metabolism that generates considerable heat. For this reason, development of fluorescent probes to detect mitochondrial temperature has become an attractive topic. Previous efforts have successfully addressed the major issues, such as temperature sensitivity and mitochondrial targetability. However, there remains a key obstacle to practical applications. Considering the highly dynamic features of mitochondria, especially the variation of the inner-membrane potential, it is quite necessary to permanently immobilize a temperature probe in mitochondria in order to avoid unstable intracellular localization along with the changes of mitochondrial status. Herein, we report Mito-TEM, the first fixable, fluorescent molecular thermometer. Mito-TEM is based on a positively charged rhodamine B fluorophore that has the tendency of being attracted to mitochondria, which have negative potential. This fluorophore containing rotatable substituents also contributes to the temperature-responsive fluorescence property. Most importantly, a benzaldehyde is introduced in Mito-TEM as an anchoring unit that condenses with aminos of the protein and thus immobilizes the probe in mitochondria. The specific immobilization of Mito-TEM in mitochondria is unambiguously demonstrated in colocalization imaging. By using Mito-TEM, a method of visualizing and quantifying a temperature distribution through grayscale imaging of mitochondria is established and further applied to monitor the temperature changes of live cells under light heating and PMA stimulation.

Silver Nanowire-Based Fluorescence Thermometer for a Single Cell.

A fluorescence thermometer based on silver nanowires (AgNWs) is realized by assembling Texas Red (TR)-marked thermal-sensitive DNA stem-loops (TR-DNA stem-loop) on the surface of AgNWs. Temperature configures the structure of the TR-DNA stem-loop and resultantly adjusts the energy transfer between TR and the AgNWs, which could sensitively control the fluorescence intensity of the thermometer. The thermometer is sensitive to the temperature ranging from 30 to 40 °C with the sensitivity of 2.6%/°C. Under the assistance of laser confocal microscopy, a temperature change within a single cell was observed by the monofilament AgNW-based thermometer.

Thermal sensing in fluid at the micro-nano-scales.

Temperature is one of the most fundamental parameters for the characterization of a physical system. With rapid development of lab-on-a-chip and biology at single cell level, a great demand has risen for the temperature sensors with high spatial, temporal, and thermal resolution. Nevertheless, measuring temperature in liquid environment is always a technical challenge. Various factors may affect the sensing results, such as the fabrication parameters of built-in sensors, thermal property of electrical insulating layer, and stability of fluorescent thermometers in liquid environment. In this review, we focused on different kinds of micro/nano-thermometers applied in the thermal sensing for microfluidic systems and cultured cells. We discussed the advantages and limitations of these thermometers in specific applications and the challenges and possible solutions for more accurate temperature measurements in further studies.

Fast MR thermometry using an echo-shifted sequence with simultaneous multi-slice imaging.

Real-time monitoring is important for the safety and effectiveness of high-intensity focused ultrasound (HIFU) therapy. Magnetic resonance imaging is the preferred imaging modality for HIFU monitoring, with its unique capability of temperature imaging. For real-time temperature imaging, higher temporal resolution and larger spatial coverage are needed. In this study, a sequence based on the echo-shifted RF-spoiled gradient echo (GRE) with simultaneous multi-slice (SMS) imaging was designed for fast temperature imaging. A phantom experiment was conducted to evaluate the accuracy of the echo-shifted sequence using a fluorescent fiber thermometer as reference. The temperature uncertainty of the echo-shifted sequence was compared with the traditional GRE sequence at room temperature through the ex vivo porcine muscle. Finally, the ex vivo porcine liver tissue experiment using HIFU heating was performed to demonstrate that the spatial coverage was increased without decreasing temporal resolution. The echo-shifted sequence had a better temperature uncertainty performance compared with the traditional GRE sequence with the same temporal resolution. The ex vivo heating experiment confirmed that by combining the SMS technique and echo-shifted sequence, the spatial coverage was increased without decreasing the temporal resolution while maintaining high temperature measurement precision. The proposed technique was validated as an effective real-time method for monitoring HIFU therapy.

DNA nanostructure-based fluorescence thermometer with silver nanoclusters

DNA nanostructure-based fluorescence thermometers were fabricated by linking fluorescent silver nanoclusters (AgNCs) and guanine-rich（G-rich）DNA chains via a thermally sensitive DNA stem-loop at terminals 5′ and 3′. Variations of temperature alter the distance between the AgNCs and G-rich DNA chain, affecting the interaction between them. As a result, the intensity of fluorescence emission from the AgNCs at 636 nm can be sensitively modulated. It was found that the intensity of such red emission is more temperature sensitive than the equivalent green emission at 543 nm; sensitivity of −3.6%/°C was achieved. Through variation of the melting temperature of the DNA stem-loop, the response temperature range of the thermometers could be readily adjusted. Novel DNA nanostructure-based fluorescence thermometers as described in this work are anticipated to be able to measure the temperature of biological systems at small scales—even a single cell.

A Cell-Targeted Non-Cytotoxic Fluorescent Nanogel Thermometer Created with an Imidazolium-Containing Cationic Radical Initiator.

A cationic fluorescent nanogel thermometer based on thermo-responsive N-isopropylacrylamide and environment-sensitive benzothiadiazole was developed with a new azo compound bearing imidazolium rings as the first cationic radical initiator. This cationic fluorescent nanogel thermometer showed an excellent ability to enter live mammalian cells in a short incubation period (10 min), a high sensitivity to temperature variations in live cells (temperature resolution of 0.02-0.84 °C in the range 20-40 °C), and remarkable non-cytotoxicity, which permitted ordinary cell proliferation and even differentiation of primary cultured cells.

A Cell‐Targeted Non‐Cytotoxic Fluorescent Nanogel Thermometer Created with an Imidazolium‐Containing Cationic Radical Initiator

AbstractA cationic fluorescent nanogel thermometer based on thermo‐responsive N‐isopropylacrylamide and environment‐sensitive benzothiadiazole was developed with a new azo compound bearing imidazolium rings as the first cationic radical initiator. This cationic fluorescent nanogel thermometer showed an excellent ability to enter live mammalian cells in a short incubation period (10 min), a high sensitivity to temperature variations in live cells (temperature resolution of 0.02–0.84 °C in the range 20–40 °C), and remarkable non‐cytotoxicity, which permitted ordinary cell proliferation and even differentiation of primary cultured cells.

Efficient mercury recovery from mercuric-thiosulfate solutions by ultraviolet photolysis

Mercury-containing wastes are hazardous due to the high toxicity and bio-accumulative effect of mercury. Examples of mercury-containing wastes include fluorescent light bulbs, thermostats, and thermometers. Recently, several aqueous-phase methods have been developed to extract mercury from mercury-containing wastes. However, the slow kinetics of mercury recovery limits the application of the aqueous-based technology. Here we designed a method of mercury recovery from a mercuric-thiosulfate solution assisted by ultraviolet photolysis. We evaluated the effect of initial pH, thiosulfate concentration and temperature on ultraviolet photolysis. The precipitation products were characterized by X-ray diffraction and X-ray photoelectron spectroscopy (XPS). Results show a mercury recovery ratio of 87.94% after ultraviolet photolysis during 240 min. We conclude that mercury can be efficiently recovered from mercuric-thiosulfate complex solutions by ultraviolet photolysis.

Developing the structure-property relationship to design solid state multi-stimuli responsive materials and their potential applications in different fields.

Prediction of multi-stimuli responsive behavior in newly developed luminogens is an appealing yet challenging puzzle, since no concrete design strategy has been developed so far. In this article, we demonstrate a potent strategy to gain a deep understanding of the structure-property relationship to design multi-stimuli responsive mechanochromic materials. To achieve our goal, a variety of new isoindolinone core based charge transfer luminogens exhibiting aggregation-induced emission (AIE) have been prepared through C-H bond activation using a cost-effective ruthenium (Ru) metal catalyzed one-pot synthetic strategy. We have shown that slight tuning of the donor moiety is found to be highly effective in controlling molecular packing and metastable energy states in solid states, and thus, optical properties and multi-stimuli responsive behaviors. The flexibility and twisting of donor moieties afford a loosely bound 'herringbone' packing, enabling reversible transformation under multiple mechanical stimuli. The cyclized derivative of the donor exhibits a completely different packing mode (i.e., cross packing), and subsequently, does not give rise to mechanochromism. The Hirshfeld surface analysis from a single crystal infers that non-covalent interactions (specifically C-H···π and π···π) are extremely important to yield mechanochromism under external force. Correlating all solid-state behavior with the molecular structure, we conclude that the synergistic effect between the twisting and conformational flexibility of donor moieties along with numerous non-covalent interactions gives rise to multi-stimuli responsive behaviors. Finally, the newly designed molecules are found to be highly emissive in solution and potentially applicable in fluorescence thermometer construction, lighting up cells, acid-base sensors and rewritable devices.

Difference in intracellular temperature rise between matured and precursor brown adipocytes in response to uncoupler and \u03b2-adrenergic agonist stimuli

Brown adipocytes function to maintain body temperature by heat production. However, direct measurement of heat production at a single cell level remains difficult. Here we developed a method to measure the temperature within primary cultured brown adipocytes using a cationic fluorescent polymeric thermometer. Placement of the thermometer within a matured brown adipocyte and a precursor cell enabled the detection of heat production following uncoupler treatment. The increase in the intracellular temperature due to stimulation with a mitochondrial uncoupler was higher in matured brown adipocytes than in precursor cells. Stimulation with a β-adrenergic receptor (β-AR) agonist, norepinephrine, raised the intracellular temperature of matured brown adipocytes to a level comparable to that observed after stimulation with a β3-AR-specific agonist, CL316.243. In contrast, neither β-AR agonist induced an intracellular temperature increase in precursor cells. Further, pretreatment of brown adipocytes with a β3-AR antagonist inhibited the norepinephrine-stimulated elevation of temperature. These results demonstrate that our novel method successfully determined the difference in intracellular temperature increase between matured brown adipocytes and precursor cells in response to stimulation by an uncoupler and β-AR agonists.

Spatially and temporally-resolved tryptophan fluorescence thermometry for monitoring the photothermal processes of gold nanorod suspensions

We constructed a confocal fluorescent thermometer to monitor the spatial and temporal temperature evolutions of gold nanorod suspensions upon continuous-wave infrared excitation. Tryptophan served as the fluorophore due to its superior temperature dependent fluorescence intensity, ca. −2%°C−1. We provided sufficient spatial resolution of 50μm and temperature sensitivity of 0.3°C. Coupled with an infrared thermocamera, we were able to obtain simultaneously the temperature evolutions in the bulk solution and on the container surface. Employment of this noncontact and optical approach allows investigation of the thermal distribution in a minute volume, less than 1mm3, without the additional perturbative factors of the mechanical thermometers, e.g., the thermal conductivity and volume of the sensor.

Supramolecular Self-Assembly Bioinspired Synthesis of Luminescent Gold Nanocluster-Embedded Peptide Nanofibers for Temperature Sensing and Cellular Imaging.

Metal nanoclusters (NCs) hold great potential as novel luminescent nanomaterials in many applications, while the synthesis of highly luminescent metal NCs still remains challenging. In this work, we report self-assembling peptides as a novel bioinspired scaffold capable of significantly enhancing the luminescence efficiency of gold nanoclusters (AuNCs). The resulting AuNCs capped with motif-designed peptides can self-assemble to form nanofiber structures, in which the luminescence of AuNCs is enhanced nearly 70-fold, with 21.3% quantum yield. The underlying mechanism responsible for the luminescence enhancement has been thoroughly investigated by the combined use of different spectroscopic and microscopic techniques. The resultant highly luminescent AuNC-decorated peptide nanofibers exhibit physicochemical properties that are advantageous for biological applications. As a proof of concept, we demonstrate the use of these nanostructure as fluorescent thermometers and for imaging living cells, both showing very promising results.

Using SiO2-Coated Gold Nanorods as Temperature Jump Photothermal Convertors Coupled with a Confocal Fluorescent Thermometer to Study Protein Unfolding Kinetics: A Case of Bovine Serum Albumin

Photoexcitation of SiO2-coated gold nanorods (AuNR@SiO2) with a 1064 nm pulse of ca. 100 μs duration leads to a temperature jump of 5 °C, as determined by the evolution of the fluorescence intensity of tryptophan. A confocal fluorescent thermometer was developed to detect the temperature evolution in a small excitation volume of ca. 10–3 mm3 and provide a spatial resolution of 100 μm. In addition, a continuous-wave laser at 1550 nm was employed to increase the initial temperature to 44 °C by heating H2O, providing alternative initial temperatures for temperature jump experiments to reveal the unfolding kinetics of bovine serum albumin (BSA). The evolution of fluorescence of BSA upon temperature jump when the stationary temperature was raised to 44 °C differed from that at room temperature, suggesting a dynamical unfolding kinetics, and a rate coefficient of 75 ± 15 s–1 was derived. In this work, we successfully demonstrated the applicability of AuNR@SiO2 as the photothermal material for the temperature ju...