Polymeric microenvironment enhancing polarity response sensitivity for discriminating lipid droplets in cancer cells.

Cellular micro-environment analysis via fluorescence probe has become a powerful method to explore the early-stage cancer diagnosis and pathophysiological process of relevant diseases. The polarity change of intracellular lipid droplets (LDs) is closely linked with disorders or diseases, which result in various physiological and pathological processes. However, the efficient design strategy for lipid droplet polarity probes with high sensitivity is lacking. To overcome this difficulty, two kinds of LDs-targeting and polarity-sensitive fluorescent probes containing carbazole and siloxane groups were rationally designed and synthesized. With the carbazole-based rotor and bridge-like siloxanes, two probes (P1 and P2) behave high sensitivity to polarity changes and show different fluorescent intensity in normal and cancer cells. Notably, polysiloxanes groups promoted the response sensitivity of the probes dramatically for the polymeric microenvironment. In addition, due to the polarity changes of LDs in cancer cells, the distinct fluorescent intensities in different channels of laser scanning confocal microscope were observed between NHA cell and U87 cells. This work could offer an opportunity to monitor the dynamic behaviors of LDs and further provide a powerful tool to be potentially applied in the early-stage diagnosis of cancer.

Polarity responsive polysiloxanes with twisting intramolecular charge transfer effect for monitoring lipophagy process and the detection of volatile organic compounds.

Polarity plays a critical role in biology and materials science, serving as a complex parameter. Imbalances in polarity within subcellular organelles are closely associated with various diseases. Moreover, volatile organic compounds (VOC) with low polarity pose significant health and safety risks, therefore, researchers have shown great interest in accurately detecting polarity. However, precise observation of polarity changes within organisms and identification of low-polarity volatile organic solvents are formidable challenges. To overcome these difficulties, we developed a versatile polymeric twisting intramolecular charge transfer (TICT) effect Polysiloxane-n (PDMS-n), utilizing polysiloxane molecular chains as "smart guides" to connect TICT molecules, inspired by the concept of "threading a needle." With the aid of PDMS-n, the process of polarity changes during cellular lipophagy was monitored in situ with high accuracy. Remarkably, the polarity changes of the local microstructure of the PDMS films were successfully visualized. PDMS-Films were also constructed, which enabled the recognition of Dichloromethane (DCM) gas during swelling. This work will contribute to the understanding of changes in cellular physiological processes, and facilitate the sensitive detection of VOCs.

Versatile Switchable Targeted Polysiloxanes for High-Resolution Visualization of Mitochondrial and Lysosomal Interactions during Ferroptosis.

Ferroptosis is an iron-dependent process that regulates cell death and is essential for maintaining normal cell and tissue survival. The explosion of reactive oxygen species characterizes ferroptosis in a significant way. Peroxynitrite (ONOO-) is one of the endogenous reactive oxygen species. Abnormal ONOO- concentrations cause damage to subcellular organelles and further interfere with organelle interactions. However, the proper conduct of organelle interactions is critical for cellular signaling and the maintenance of cellular homeostasis. Therefore, investigating the effect of ONOO- on organelle interactions during ferroptosis is a highly attractive topic. To date, it has been challenging to visualize the full range of ONOO- fluctuations in mitochondria and lysosomes during ferroptosis. In this paper, we constructed a switchable targeting polysiloxane platform. During the selective modification of NH2 groups located in the side chain, the polysiloxane platform successfully constructed fluorescent probes targeting lysosomes and mitochondria (Si-Lyso-ONOO, Si-Mito-ONOO), respectively. Real-time detection of ONOO- in lysosomes and mitochondria during ferroptosis was successfully achieved. Remarkably, the occurrence of autophagy during late ferroptosis and the interaction between mitochondria and lysosomes was observed via the differentiated responsive strategy. We expect that this switchable targeting polysiloxane functional platform will broaden the application of polymeric materials in bioimaging and provide a powerful tool for further deeper understanding of the ferroptosis process.

Hyperbranched polysiloxane-based probe with enhanced lipophilicity for visualizing ONOO- fluctuations in endoplasmic reticulum.

The endoplasmic reticulum, a cellular signaling regulator, participates in the synthesis and secretion of many proteins, glycogen, lipids and cholesterol substances. Peroxynitrite (ONOO-) is a highly oxidative and nucleophilic agent. Abnormal fluctuations of ONOO- induce oxidative stress in the endoplasmic reticulum, further disrupting the normal function of protein folding and transport and glycosylation modification, ultimately leading to neurodegenerative diseases, cancer and Alzheimer's disease. Up to now, most probes have tended to achieve targeting functions by introducing specific targeting groups. However, this approach increased the difficulty of the construction process. Therefore, a simple and efficient construction strategy for fluorescent probes with excellent specificity targeting the endoplasmic reticulum is lacking. To overcome this difficulty and put forward an efficient design strategy for the endoplasmic reticulum targeted probes, in this paper, we constructed alternating rigid and flexible polysiloxane-based hyperbranched polymeric probes (Si-Er-ONOO) by bonding perylenetetracarboxylic anhydride and silicon-based dendrimers for the first time. Efficient and specific targeting of the endoplasmic reticulum was successfully achieved by the excellent lipid solubility of Si-Er-ONOO. Furthermore, we observed different effects of metformin and rotenone on the changes of ONOO- volatility in the cellular and zebrafish internal environment by Si-Er-ONOO. We believe that Si-Er-ONOO will expand the application of organosilicon hyperbranched polymeric materials in bioimaging and provide an excellent indicator of reactive oxygen species fluctuations in biological systems.

A novel coumarin-TPA based fluorescent probe for turn-on hypochlorite detection and lipid-droplet-polarity bioimaging in cancer cells.

A novel fluorescent compound, named C-TPA, based on coumarin (acceptor) and triphenylamine (donor) was facilely designed and fabricated through a one-step Suzuki coupling reaction. As a donor group, triphenylamine can efficiently enhance the fluorescence intensity and photostability of coumarin, and thus improve the detection efficiency. C-TPA-S was obtained from C-TPA treated with Lawesson's reagent and C-TPA-S can be used for the turn-on detection of hypochlorite through oxidative desulfurization with a low detection limit of 0.12 µM. Moreover, the intramolecular charge transfer process between the donor and acceptor group endows C-TPA with solvatochromism property and makes C-TPA a good candidate for polarity detection. The C-TPA with bright green fluorescence was highly efficient for imaging the microenvironment of polarity both in living cells and tissues with high selectivity and photostability, which can be applied in the diagnosis for the cancer cells.

Multistimuli-responsive fluorescent probes based on spiropyrans for the visualization of lysosomal autophagy and anticounterfeiting.

Lysosomes, as the main degradative organelles, play an important role in a variety of cellular metabolic activities including autophagy and apoptosis, catabolism and transporting substances. Lysosomal autophagy is an important physiological process and causes a slight change in the intra-lysosomal pH to facilitate the breakdown of macromolecular proteins. Therefore, detecting the fluctuation of intra-lysosomal pH is of great significance in monitoring physiological and pathological activities in living organisms. However, few probes have enabled the ratiometric monitoring of lysosomal pH and lysosomal autophagy in dual channels. Fortunately, spiropyrans, as compounds with multistimuli-responsive discoloration properties, form two different isomers under acid induction and ultraviolet induction. To fill this gap, in this work, two novel multistimuli-responsive fluorescent probes with lysosomal targeting in dual channels based on spiropyrans were rationally designed and synthesized. Notably, the two probes exhibited different absorption wavelengths in their UV-responsive and pH-responsive moieties due to their different electron-donating groups. Moreover, bioimaging experiments clearly demonstrate that the probes Lyso-SP and Lyso-SQ monitor lysosomal autophagy by facilitating the visualization of fluctuations in intra-lysosomal pH. Meanwhile, their potential applications in the field of dual-anticounterfeiting were explored based on their photoluminescence ability. We expect that more multistimuli-responsive fluorescent probes can be developed by this design approach.

A fluorescent probe based on POSS for facilitating the visualization of HClO and NO in living cells and zebrafish.

The main production area of HClO and NO is the mitochondria and has modulatory effects on multiple human diseases. Simultaneous detection of signaling molecules such as HClO and NO is an important approach for exploring the complex relationship between HClO and NO in mitochondria. However, most probes can detect only one species or are unable to complete the monitoring of HClO and NO in the NIR channel. There are only few reports on reasonable tools that can simultaneously monitor the presence of HClO and NO in the NIR channel. In this work, to solve this difficulty, a POSS-assisted NIR fluorescent probe with dual-response was rationally devised and developed. The probe Mito-Cy possessed high specificity and responsiveness to HClO and NO in spectral experiments. Notably, the probe exhibited excellent responsiveness and sensitivity to HClO and NO in living cells and the zebrafish model.

Hot-Band Absorption of a Cationic RNA Probe Enables Visualization of ΔΨm via the Controllable Anti-Stokes Shift Emission.

Mitochondrial membrane potential (ΔΨm) is an important biophysical parameter playing central roles in cell apoptosis, mitochondrial dysfunction, and other biological and pathological processes. Herein, we have rationally designed and fabricated a unique fluorescent probe for convenient ΔΨm visualization based on hot-band absorption and controllable anti-Stokes shift emission. The robust probe was excitable via hot-band absorption and emitted anti-Stokes upconversion emission and Stokes downconversion fluorescence simultaneously. The anti-Stokes emission could be efficiently inhibited upon the binding to RNA. The cationic probe targeted mitochondria in living cells with high ΔΨm and displayed both anti-Stokes green emission and ordinary red fluorescence. After the decrease of ΔΨm, the probe immigrated out of mitochondria to RNA and nucleolus, which showed only red emission owing to the inhibition of anti-Stokes fluorescence. In this manner, the ΔΨm could be visualized in dual-color mode. The probe enabled clearly monitoring the reversible changes in ΔΨm and was successfully employed to visualize oxidative damage of living cells. The decrease of ΔΨm in living tissues was also successfully observed with the newly designed probe.

The diversity of the coordination bond generated a POSS-based fluorescent probe for the reversible detection of Cu(II), Fe(III) and amino acids.

In recent years, it has been found that Cu2+, Fe3+, and amino acids play an irreplaceable and subtle role in organisms and have attracted the considerable attention of many researchers. Therefore, it is vital to design visual indicators to reveal the relationships between metal ions and amino acids. However, there have been few reports on this vigorous subject. Fortunately, based on the different coordination effects between metal ions and boron groups, we have designed an accessible fluorescent probe (PSI-A). Borane was introduced as an ion-sensitive group to form a novel POSS-based fluorescent probe, which achieves fascinating performance, in situ dynamic multiple detection, excellent photostability, and enervative biological toxicity. PSI-A exhibited predominant selectivity and sensitivity to Cu2+/amino acids and Fe3+/amino acids sequence reactions in HepG2 cells and zebrafish. The fluorescence of PSI-A was quenched by Cu2+, which can be recovered by adding Asp, Ser, Arg, Ace or Trp. Additionally, the fluorescence of PSI-A quenched by Fe3+ can be restored after adding Asp. PSI-A is available to monitor Cu2+/amino acids and Fe3+/amino acids sequence reactions and can be repeated for at least three consecutive cycles without a fatigued performance. Therefore, this multifunctional fluorescent probe may have prospective application potentials in the biological field.

A POSS-assisted fluorescent probe for the rapid detection of HClO in mitochondria with a large emission wavelength in dual channels.

Hypochlorous acid (HClO) is closely related to many diseases and is an inevitable part of the physiological processes. It is significant to detect HClO in mitochondria for getting meaningful physiological and pathological information. However, adequate tools to detect HClO with emissions in two channels are rarely reported. To achieve this target, in this work, a "turn-off" visual and near infrared (NIR) fluorescent dual emission probe D6 based on polyhedral oligomeric silsesquioxanes (POSS) was successfully designed and synthesized. D6 showed high selectivity and sensitivity to HClO. Notably, the emission wavelength of D6 reached 820 nm due to the assistance of the POSS cage. In addition, bioimaging experiments clearly showed that probe D6 promoted the visualization of exogenous and endogenous HClO in living HepG2 cells and zebrafish models.

Step-wise functionalization of polysiloxane towards a versatile dual-response fluorescent probe and elastomer for the detection of H2S in two-photon and NO in near-infrared modes.

Simultaneous detection of gasotransmitters such as NO and H2S is a crucial task to investigate the complicated relationship between NO and H2S. In this work, we presented a new approach to construct a dual response fluorescent probe for NO/H2S by step-wise functionalization of polysiloxanes. A near-infrared (NIR) and two-photon (TP) fluorescent probe, P-CYN, was fabricated for respectively visualizing H2S and NO in vivo. This work presented a powerful tool for the investigation of the relationship between gasotransmitters, and provided a valid strategy for further design of dual-response fluorescent probes.

A novel polythioether-based rhodamine B fluorescent probe via successive click reaction and its application in iron ion detection and cell imaging.

Polythioether has good chemical stability and biocompatibility and is a kind of promising polymers for the application of optical materials, medical materials and energy conversion materials. However, the fluorescent probe based on polythioether is still rare. Herein, a series of polythioether based polymer fluorescent probes were synthesized by successive thiol click reaction under ultraviolet light at room temperature. The poly(thioether)s have good selectivity and responsiveness to iron ions and can be applied in cell imaging, which indicate that the broad application prospects of polythioether-based fluorescent probes in ion detection and bioimaging.

AIE-active polysiloxane-based fluorescent probe for identifying cancer cells by locating lipid drops.

Comparing with normal cells, Lipid droplets (LDs) of cancer cells show lower polarity and less quantity, which can be utilized as a marker for cancer diagnosis. However, the investigation of LDs in living cancer cells is restricted by the lack of effective molecular tools. Herein, we first reported a novel polysiloxane-based polymer fluorescent polar probe TR-1 with AIE properties, which realized the possibilities for locating LDs. It can aggregate in the LDs of cancer cells and show a stronger fluorescent signal to conduct cancer diagnosis. Moreover, the excellent photostability of TR-1 enable stable fluorescence to exhibit in cancer cells during effective time.

Novel fluorene-based fluorescent probe with excellent stability for selective detection of SCN- and its applications in paper-based sensing and bioimaging.

SCN- is one of the most important anions in metabolic processes. However, the investigation of SCN- in living systems is restricted by the lack of stable functional molecular tools. Herein, the first fluorene-based polymer fluorescent probe V1 was synthesized through rational design. Compared with small molecule fluorescent probes, V1 exhibited excellent fluorescence stability in bovine serum albumin (BSA) solution. Furthermore, the V1-based paper sensor was highly selective toward SCN- in aqueous solution. Significantly, these merits of the probe V1 enable the detection of SCN- in different living cell lines and zebrafish.

Novel fluorescent probe with a bridged Si-O-Si bond for the reversible detection of hypochlorous acid and biothiol amino acids in live cells and zebrafish.

Herein, we report the design of a novel fluorescent probe consisting of a naphthalimide fluorophore and a silicone small molecule for the reversible detection of hypochlorous acid and biothiol amino acids. The response mechanism of BSi-1 is based on the concept of the S-based oxidation/reduction. The probe was found to be suitable for imaging HOCl in HeLa, RAW 264.7 cells and zebrafish, demonstrating its utility in biological applications.

Synthesis of Silane-Based Poly(thioether) via Successive Click Reaction and Their Applications in Ion Detection and Cell Imaging.

A series of poly(thioether)s containing silicon atom with unconventional fluorescence were synthesized via successive thiol click reaction at room temperature. Although rigid π-conjugated structure did not exist in the polymer chain, the poly(thioether)s exhibited excellent fluorescent properties in solutions and showed visible blue fluorescence in living cells. The strong blue fluorescence can be attributed to the aggregation of lone pair electron of heteroatom and coordination between heteroatom and Si atom. In addition, the responsiveness of poly(thioether) to metal ions suggested that the selectivity of poly(thioether) to Fe3+ ion could be enhanced by end-modifying with different sulfhydryl compounds. This study further explored their application in cell imaging and studied their responsiveness to Fe3+ in living cells. It is expected that the described synthetic route could be extended to synthesize novel poly(thioether)s with superior optical properties. Their application in cell imaging and ion detection will broaden the range of application of poly(thioether)s.

Pyrenyl-Functionalized Polysiloxane Based on Synergistic Effect for Highly Selective and Highly Sensitive Detection of 4-Nitrotoluene.

4-Nitrotoluene (NT) is an important intermediate for the manufacture of dyes, agricultural chemicals, medicaments, and synthetic fibers. But its wide use has resulted in a series of ecological problems and health issues due to high toxicity, mutagenicity, and carcinogenicity. However, no fluorescent probes with high selectivity and high sensitivity to NT has been reported yet, and the current probes usually prefer to detect multi-nitrosubstituted nitroaromatic compounds (NACs) including 2,4,6-trinitrotoluene and 2,4,6-trinitrophenol. Herein, we report a series of pyrene-functionalized polysiloxanes with high selectivity and high sensitivity to NT. Pyrene was introduced into polysiloxane through a carbon-carbon double bond, and the formed rigid side-chain fluorophore and flexible backbone structure of polysiloxane provide an ideal platform for the highly selective detection of NT. We further explored the possible response mechanism and speculated that the high selectivity and high sensitivity were derived from the synergistic effect between steric hindrance and dipolar interaction. In addition, paper sensors based on the obtained fluorescent materials were fabricated and change in their high sensitivity and visible fluorescence indicated that the paper sensor is a simple testing tool for portable and visual detection of NT. Furthermore, the design strategy in this work provides a novel synthetic route to synthesize other fluorescent probes with unique selectivity to NACs detection.

Novel polysiloxane-based rhodamine B fluorescent probe for selectively detection of Al3+ and its application in living-cell and zebrafish imaging.

Polysiloxanes have excellent stability and biological relevance and are suitable for biological research. However, there were few polysiloxane-based fluorescent probes for bioimaging. This report successfully designed a new polysiloxane-based polymer fluorescent probe (RB-1) for the first time as a "turn-on" fluorescent probe response to Al3+ ion with highly sensitive and selectivity. Importantly, this probe could also apply both in cell and zebrafish imaging, indicating the huge application development prospects of polysiloxane-based fluorescent probes in future.

Binding Reaction Sites to Polysiloxanes: Unique Fluorescent Probe for Reversible Detection of ClO-/GSH Pair and the in Situ Imaging in Live Cells and Zebrafish.

PDMS is biocompatible, economically viable, transparent, and facile to handle and thus is suitable for fluorescent microscopy and biological research. However, there has been no report about polysiloxane-based fluorescent probes applied in bioimaging. In this report, a two-photon polysiloxane-based reversible luminescent probe (P1) was fabricated for the first time. P1 is a powerful tool for detecting the ClO-/GSH cycle in situ both in live cells and in zebrafish. This work demonstrates the potential of polysiloxane-based fluorescent probes for versatile in vivo or in vitro applications in the future.

Thermally Responsive Materials for Bioimaging.

The last decade has witnessed multiple thermally responsive materials emerge as a significant class of stimuli-responsive materials. These materials are elaborately designed and exert interesting properties. Herein, an overview of thermally responsive materials with respect to design strategies, fabrication procedures, and their applications is presented. Recently reported thermally responsive materials are highlighted. Then, applications of thermally responsive materials in bioimaging are summarized.