NIR Fluorescent Dye Research Articles

Steric protection of near-infrared fluorescent dyes for enhanced bioimaging.

Near-fluorescent (NIR) dyes that absorb and emit light in the wavelength range of 650-1700 nm are well-suited for bioimaging due to the improved image contrast and increased penetration of the long-wavelength light through biological tissue. However, the imaging performance of NIR fluorescent dyes is limited by several inherent photophysical and physicochemical properties including, low fluorescence quantum yield, high chemical and photochemical reactivity, propensity to self-aggregate in water, non-specific association with off-target biological sites, and non-optimal pharmacokinetic profiles in living subjects. In principle, all these drawbacks can be alleviated by steric protection which is a structural process that surrounds the fluorophore with bulky groups that block undesired intermolecular interactions. The literature methods to sterically protect a long-wavelength dye can be separated into two general strategies, non-covalent dye encapsulation and covalent steric appendage. Illustrative examples of each method show how steric protection improves bioimaging performance by providing: (a) increased fluorescence brightness, (b) higher fluorophore ground state stability, (c) decreased photobleaching, and (d) superior pharmacokinetic profile. Some sterically protected dyes are commercially available and further success with future systems will require experts in chemistry, microscopy, cell biology, medical imaging, and clinical medicine to work closely as interdisciplinary teams.

Cluster Preface: Functional Dyes

Youjun Yang (left) received his BS (2002) from the University of Science and Technology of China, and his PhD (2007) with Prof. Robert M. Strongin at Louisiana State University. Then, he joined the Anslyn group at UT Austin as a postdoc. In 2010, he joined the faculty of the school of pharmacy, East China University of Science and Technology (ECUST). His research interests include NIR fluorescent dyes, photo-responsive dyes, and fluorescent molecular probes. Jiangli Fan (right) obtained her PhD at Dalian University of Technology (2005). Now she is a Professor at the School of Chemical Engineering at Dalian University of Technology, and she received the Distinguished Young Scholars of NSFC (2019). Her research interests mainly focus on the development of functional dyes for biomedical analysis and disease treatment.

Enlarging the Stokes Shift by Weakening the π-Conjugation of Cyanines for High Signal-to-Noise Ratiometric Imaging.

The signal-to-noise ratio (SNR) is one of the key features of a fluorescent probe and one that often defines its potential utility for in vivo labeling and analyte detection applications. Here, it is reported that introducing a pyridine group into traditional cyanine-7 dyes in an asymmetric manner provides a series of tunable NIR fluorescent dyes (Cy-Mu-7) characterized by enhanced Stokes shifts (≈230nm) compared to the parent cyanine 7 dye (<25nm). The observed Stokes shift increase is ascribed to symmetry breaking of the Cy-Mu-7 core and a reduction in the extent of conjugation. The fluorescence signals of the Cy-Mu-7 dyes are enhanced upon confinement within the hydrophobic cavity of albumin or via spontaneous encapsulation within micelles in aqueous media. Utilizing the Cy-Mu-7, ultra-fast in vivo kidney labeling in mice is realized, and it is found that the liver injury will aggravate the burden of kidney by monitoring the fluorescence intensity ratio of kidney to liver. In addition, Cy-Mu-7 could be used as efficient chemiluminescence resonance energy transfer acceptor for the reaction between H2 O2 and bisoxalate. The potential utility of Cy-Mu-7 is illustrated via direct monitoring fluctuations in endogenous H2 O2 levels in a mouse model to mimic emergency room trauma.

NIR Fluorescence lifetime macroscopic imaging with a time-gated SPAD camera.

The performance of SwissSPAD2 (SS2), a large scale, widefield time-gated CMOS SPAD imager developed for fluorescence lifetime imaging, has recently been described in the context of visible range and fluorescence lifetime imaging microscopy (FLIM) of dyes with lifetimes in the 2.5 - 4 ns range. Here, we explore its capabilities in the NIR regime relevant for small animal imaging, where its sensitivity is lower and typical NIR fluorescent dye lifetimes are much shorter (1 ns or less). We carry out this study in a simple macroscopic imaging setup based on a compact NIR picosecond pulsed laser, an engineered diffuser-based illumination optics, and NIR optimized imaging lens suitable for well-plate or small animal imaging. Because laser repetition rates can vary between models, but the synchronization signal frequency accepted by SS2 is fixed to 20 MHz, we first checked that a simple frequency-division scheme enables data recording for different laser repetition rates. Next, we acquired data using different time gate widths, including gates with duration longer than the laser period, and analyzed the resulting data using both standard nonlinear least-square fit (NLSF) and phasor analysis. We show that the fixed synchronization rate and large gate widths characterizing SS2 (10 ns and over) are not an obstacle to accurately extracting lifetime in the 1 ns range and to distinguishing between close lifetimes. In summary, SS2 and similar very large gated SPAD imagers appear as a versatile alternative to other widefield time-resolved detectors for NIR fluorescence lifetime imaging, including preclinical molecular applications.

The Design and Bioimaging Applications of NIR Fluorescent Organic Dyes with High Brightness

AbstractDue to the merits of decreased photon attenuation, autofluorescence, and scattering, the near‐infrared (NIR, 700–1700 nm) region is an important window in the field of biomedicine, such as in vivo fluorescence imaging, in which both the optical detection depth and the resolution/contrast have been significantly improved. In particular, for second NIR (NIR‐II, 1000–1700 nm) dyes, biological tissues have almost no background interference. Typical fluorophores have excellent spectral performance and rich functional modification sites and can be optimized into NIR dyes for bioimaging. However, as the absorption/emission wavelength of fluorophores redshift to NIR, it is challenging to keep fluorophores with satisfying brightness. Therefore, for the purpose of increasing the absorption/emission wavelength of dye while promoting its brightness, it is necessary to study the structure–property relationship of the dyes. This review introduces the influences of fluorophores’ structure on their photophysical properties, summarizes the strategies for maintaining high fluorescence brightness along with redshifted absorption/emission wavelengths, and the latest advances of highly fluorescent brightness dyes. Finally, the opportunities and challenges in this emerging field are also provided. The authors aim to provide insightful design guidelines and clear overview of highly bright NIR fluorescent dyes, which might trigger new ideas and applications.

Manganese-based multifunctional nanoplatform for dual-modal imaging and synergistic therapy of breast cancer

Manganese has recently been exploited for cancer immunotherapy, fenton-like reaction-mediated chemo-dynamic therapy, and magnetic resonance imaging. The integration of multiple roles of manganese into one platform is of great significance for cancer theranostics and tumor inhibition. Here, we designed a multifunctional nanoplatform based on manganese, which consisted of a manganese-containing inner core and a phospholipid bilayer shell co-loaded with glucose oxidase (GOx), paclitaxel (PTX), and a NIR fluorescent dye (NanoMn-GOx-PTX). In a pH-dependent manner, the nanoplatform released manganese ions and payloads inside the tumor cells. In vitro characterization and cellular experiments indicated that NanoMn-GOx-PTX could catalyze the conversion of glucose into reactive oxygen species (ROS) through a cascade Fenton-like reaction as well as release free PTX. The consumption of glucose, ROS production, and the chemotherapeutic effect of PTX contributed to the superior cytotoxicity and apoptosis of 4T1 cancer cells. Moreover, NanoMn-GOx-PTX effectively induced the production of large amounts of type I interferon and pro-inflammatory cytokines in vivo, activating the innate immune response. Through the synergistic functions of the above components, NanoMn-GOx-PTX exerted the strongest anti-tumor effect in 4T1 tumor-bearing models. Therefore, the manganese-based nanoplatform could serve as a promising theranostic tool for breast cancer therapy. Statement of significance1) This nanoplatform can be used as a universal tool for delivering proteins and anticancer drugs into cells; 2) The PEG-modified phospholipid bilayer shell plays a significant role in retarding the release of overloaded manganese ions and drugs in a pH-sensitive manner; 3) The released Mn2+ has the ability to enhance T1 contrast in magnetic resonance imaging; 4) The released Mn2+ can function as nanoadjuvants to activate the cGAS-STING pathway and effectively induce the natural immune response;5) The overloaded manganese ions are combined with glucose oxidase to form a cascade reaction system, indirectly converting glucose into ROS to induce oxidative damage of tumor tissue.

Synthesis of a NIR fluorescent dye and its application for rapid detection of HSO3− in living cells

SO2 and its derivatives (HSO3−, SO32−) play a significant important role in many industries and organisms. Based on coumarin and benzopyranose, a promising NIR ratiometric fluorescent probe BAOA (11-oxo-2,2′,3,3′,6,7,7′,8′-octahydro -1H,1′H,5H,6′H,11H- [10,12′-bipyrano [2,3-f] pyrido [3,2,1-ij] quinolin]-13′-ium perchlorate) was designed and developed to detect HSO3− rapidly and sensitively. The sensing mechanism was Michael addition reaction, in which, strongly nucleophilic HSO3− attacked carbon-carbon double bonds and BAOA-HSO3 was formed. A superior linear calibration curve between the fluorescence ratio I490/I722 and concentrations of HSO3− was obtained in the range of 1.25–8.75 μmol/L and the LOD was figured out as 63.0 nmol/L. Cell experiments showed that BAOA could not only locate mitochondria, lysosomes, ER and Golgi but also detect exogenous HSO3− in living cells.

Colorimetric and Ratiometric Fluorescence Detection of HSO3- With a NIR Fluorescent Dye.

Bisulfite (HSO3-) has been widely used in food and industry, which has brought convenience to human life, but also seriously endangered human health. In this work, the probe PBI was designed and synthesized to detect bisulfite (HSO3-) through nucleophilic addition reaction. The probe PBI showed a selective reaction to HSO3- and can quantitatively detect HSO3-. At the same time, the color of the probe PBI changed significantly, which provided a simple method for the naked eye to identify HSO3-. Finally, it was successfully applied to the fluorescence imaging of HSO3- in living cells.

Exploiting a Rose Bengal-bearing, oxygen-producing nanoparticle for SDT and associated immune-mediated therapeutic effects in the treatment of pancreatic cancer

Sonodynamic therapy (SDT) is an emerging stimulus-responsive approach for the targeted treatment of solid tumours. However, its ability to generate stimulus-responsive cytotoxic reactive oxygen species (ROS), is compromised by tumour hypoxia. Here we describe a robust means of preparing a pH-sensitive polymethacrylate-coated CaO2 nanoparticle that is capable of transiently alleviating tumour hypoxia. Systemic administration of particles to animals bearing human xenograft BxPC3 pancreatic tumours increases oxygen partial pressures (PO2) to 20–50 mmHg for over 40 min. RT-qPCR analysis of expression of selected tumour marker genes in treated animals suggests that the transient production of oxygen is sufficient to elicit effects at a molecular genetic level. Using particles labelled with the near infra-red (nIR) fluorescent dye, indocyanine green, selective uptake of particles by tumours was observed. Systemic administration of particles containing Rose Bengal (RB) at concentrations of 0.1 mg/mg of particles are capable of eliciting nanoparticle-induced, SDT-mediated antitumour effects using the BxPC3 human pancreatic tumour model in immuno-compromised mice. Additionally, a potent abscopal effect was observed in off-target tumours in a syngeneic murine bilateral tumour model for pancreatic cancer and an increase in tumour cytotoxic T cells (CD8+) and a decrease in immunosuppressive tumour regulatory T cells [Treg (CD4+, FoxP3+)] was observed in both target and off-target tumours in SDT treated animals. We suggest that this approach offers significant potential in the treatment of both focal and disseminated (metastatic) pancreatic cancer.

Room-Temperature Phosphorescence Resonance Energy Transfer for Construction of Near-Infrared Afterglow Imaging Agents.

Afterglow imaging that detects photons after cessation of optical excitation avoids tissue autofluorescence and thus possesses higher sensitivity than traditional fluorescence imaging. Purely organic molecules with room-temperature phosphorescence (RTP) have emerged as a new library of benign afterglow agents. However, most RTP luminogens only emit visible light with shallow tissue penetration, constraining their in vivo applications. This study presents an organic RTP nanoprobe (mTPA-N) with emission in the NIR range for in vivo afterglow imaging. Such a probe is composed of RTP molecule (mTPA) as the phosphorescent generator and an NIR-fluorescent dye as the energy acceptor to enable room-temperature phosphorescence resonance energy transfer (RT-PRET), ultimately resulting in redshifted phosphorescent emission at 780nm. Because of the elimination of background noise and redshifted afterglow luminescence in a biologically transparent window, mTPA-N permits imaging of lymph nodes in living mice with a high signal-to-noise ratio. This study thus opens up a universal approach to develop organic RTP luminogens into NIR afterglow imaging agents via construction of RT-PRET.

Red/NIR Thermally Activated Delayed Fluorescence from Aza-BODIPYs.

The search for long-lived red and NIR fluorescent dyes is challenging and hitherto scarcely reported. Herein, the viability of aza-BODIPY skeleton as a promising system for achieving thermal activated delayed fluorescent (TADF) probes emitting in this target region is demonstrated for the first time. The synthetic versatility of this scaffold allows the design of energy and charge transfer cassettes modulating the stereoelectronic properties of the energy donors, the spacer moieties and the linkage positions. Delayed emission from these architectures is recorded in the red spectral region (695-735 nm) with lifetimes longer than 100 μs in aerated solutions at room temperature. The computational-aided photophysical study under mild and hard irradiation regimes disclose the interplay between molecular structure and photonic performance to develop long-lived fluorescence red emitters through thermally activated reverse intersystem crossing. The efficient and long-lasting NIR emission of the newly synthesized aza-BODIPY systems provides a basis to develop advanced optical materials with exciting and appealing photonic response.

P-Selectin-Based Dual-Model Nanoprobe Used for the Specific and Rapid Visualization of Early Detection toward Severe Acute Pancreatitis in Vivo.

Identifying severe acute pancreatitis (SAP) as soon as possible is critical for achieving optimal outcomes and saving lives. In this study, a novel P-selectin-targeted, NIR fluorescent dye (Cy 5.5)-labeled dual-modal nanoprobe based on diethylenetriaminepentaacetic chelates (Gd-DTPA-Cy5.5-PsLmAb) was constructed for the bimodal imaging of SAP at the early stage. Gd-DTPA-Cy5.5-PsLmAb was prepared, and its structure was characterized by Fourier transform infrared spectroscopy, UV-vis spectroscopy, and fluorescence spectroscopy, and its stability was evaluated. Biocompatibility was evaluated by the hemolysis and cytotoxicity assays. The enzyme-linked immunosorbent assay was used to detect and evaluate the expression of P-selectin in the peripheral blood of 11 patients with acute pancreatitis (AP) and 5 healthy volunteers. The bimodal imaging ability of Gd-DTPA-Cy5.5-PsLmAb nanoprobes was evaluated via near-infrared fluorescence (NIRF) and magnetic resonance imaging (MRI) in AP animal models in vivo. Gd-DTPA-Cy5.5-PsLmAb showed low toxicity to human embryonic kidney cells (293T cells) and good blood compatibility. The P-selectin levels of humans and rats in the mild acute pancreatitis (MAP)/SAP stage were significantly higher than those in the control group and reached the highest level at the SAP stage. Furthermore, Gd-DTPA-Cy5.5-PsLmAb nanoprobes showed clear NIRF imaging of mouse pancreas at the MAP stage and SAP stage by a fluorescence signal at 6.09 × 108 and 1.95 × 109, respectively. Meanwhile, Gd-DTPA-Cy5.5-PsLmAb nanoprobes also successfully showed the T1-weighted MR signal of rat pancreas at the MAP stage, but Gd-DTPA seldom showed any signal increase at the MAP stage; Gd-DTPA-Cy5.5-PsLmAb and Gd-DTPA could show an increasing MR signal of rat pancreas at the SAP stage. Gd-DTPA-Cy5.5-PsLmAb proved to offer a stronger signal than Gd-DTPA.Our findings indicate that Gd-DTPA-Cy5.5-PsLmAb is an effective and specific MR/NIRF dual nanoprobe for bimodal imaging, providing a promising diagnostic approach for early SAP in clinic.

Real‐Time In Vivo Detection of Cellular Senescence through the Controlled Release of the NIR Fluorescent Dye Nile Blue

AbstractIn vivo detection of cellular senescence is accomplished by using mesoporous silica nanoparticles loaded with the NIR‐FDA approved Nile blue (NB) dye and capped with a galactohexasaccharide (S3). NB emission at 672 nm is highly quenched inside S3, yet a remarkable emission enhancement is observed upon cap hydrolysis in the presence of β‐galactosidase and dye release. The efficacy of the probe to detect cellular senescence is tested in vitro in melanoma SK‐Mel‐103 and breast cancer 4T1 cells and in vivo in palbociclib‐treated BALB/cByJ mice bearing breast cancer tumor.

Real-Time In Vivo Detection of Cellular Senescence through the Controlled Release of the NIR Fluorescent Dye Nile Blue.

In vivo detection of cellular senescence is accomplished by using mesoporous silica nanoparticles loaded with the NIR-FDA approved Nile blue (NB) dye and capped with a galactohexasaccharide (S3). NB emission at 672 nm is highly quenched inside S3, yet a remarkable emission enhancement is observed upon cap hydrolysis in the presence of β-galactosidase and dye release. The efficacy of the probe to detect cellular senescence is tested in vitro in melanoma SK-Mel-103 and breast cancer 4T1 cells and in vivo in palbociclib-treated BALB/cByJ mice bearing breast cancer tumor.

Bright near-infrared chemiluminescent dyes: Phthalhydrazides conjugated with fluorescent BODIPYs

A series of phthalhydrazides conjugated with NIR fluorescent BODIPY dyes were synthesized (three steps, overall yields 68–83%) and found to show strong NIR chemiluminescence (CL) with maxima in the range of 670–736 nm. One of these showed a CL quantum yield reaching 8.4 times higher than that of luminol. Tuning of the fluorescence band of the BODIPY unit directly leads to a red-shifted CL, developing a strategy for designing NIR chemiluminogens.

Synthesis, vibrational spectroscopy and X-ray structural characterization of novel NIR emitter squaramides.

Two new 2-naphthyl squaramides, 3-methoxy -4-(2-naphtalenylamino)-3-cyclobutene-1,2-dione (SQ-NPh1) and bis-3,4-(2-naphtalenylamino)-3-cyclobutene-1,2-dione (SQ-NPh2) were synthesized via condensation reaction between the dimethylsquarate and 2-naphthylamine. The spectrometric characterization by 13C NMR confirmed the obtaining of the squaramide derivative and nor the squaraine analog. This hypothesis was corroborated by Raman and Infrared spectroscopy since the characteristic vibrational bands related to the oxocarbon portion of both structures have been assigned, such as the ones for SQ-NPh1 and SQ-NPh2. The single-crystal X-ray crystallography for SQ-NPh1 crystal structures have been solved and the structure of SQ-NPh2 have been refined using Powder Diffraction state-of-art. The SQ-NPh1 crystallizes in monoclinic system in P2/c space group. Both squaramides present absorption in the ultra-visible (220-370 nm) and fluorescent emission in the near-infrared (780-800 nm), besides they also presented high thermal stability (around 570 °C). Generally, only squaraines are reported as NIR emitters, this is the first description of NIR emission for squaramides, and since the synthesis of squaramides is very easy and the rational design of small-molecule NIR fluorophores is of high priority and great value, these results are very promising for the development of novel NIR fluorescent dyes.

Biodegradable and pH-Responsive Acetalated Dextran (Ac-Dex) Nanoparticles for NIR Imaging and Controlled Delivery of a Platinum-Based Prodrug into Cancer Cells.

Nanoparticles (NPs) based on the biodegradable acetalated dextran polymer (Ac-Dex) were used for near-infrared (NIR) imaging and controlled delivery of a PtIV prodrug into cancer cells. The Ac-Dex NPs loaded with the hydrophobic PtIV prodrug 3 (PtIV/Ac-Dex NPs) and with the novel hydrophobic NIR-fluorescent dye 9 (NIR-dye 9/Ac-Dex NPs), as well as Ac-Dex NPs coloaded with both compounds (coloaded Ac-Dex NPs), were assembled using a single oil-in-water nanoemulsion method. Dynamic light scattering measurements and scanning electron microscopy images showed that the resulting Ac-Dex NPs are spherical with an average diameter of 100 nm, which is suitable for accumulation in tumors via the enhanced permeation and retention effect. The new nanosystems exhibited high drug-loading capability, high encapsulation efficiency, high stability in physiological conditions, and pH responsiveness. Drug-release studies clearly showed that the PtIV prodrug 3 release from Ac-Dex NPs was negligible at pH 7.4, whereas at pH 5.5, this compound was completely released with a controlled rate. Confocal laser scanning microscopy unambiguously showed that the NIR-dye 9/Ac-Dex NPs were efficiently taken up by MCF-7 cells, and cytotoxicity assays against several cell lines showed no significant toxicity of blank Ac-Dex NPs up to 1 mg mL-1. The IC50 values obtained for the PtIV prodrug encapsulated in Ac-Dex NPs were much lower when compared with the IC50 values obtained for the free PtIV complex and cisplatin in all cell lines tested. Overall, our results demonstrate, for the first time, that Ac-Dex NPs constitute a promising drug delivery platform for cancer therapy.

Abstract 3729: Tumor-specific antibody labeling of pancreatic cancer in a patient-derived orthotopic xenograft (PDOX) mouse model using a fluorescent humanized anti-CEA antibody

Abstract Introduction: Delineation of tumor margins is critical in oncologic surgery, particularly in resection of pancreatic cancer. Surgeons are limited in visualization with bright light surgery. Fluorescence guided surgery (FGS) can help surgeons better visualize all potential cancer cells during the operation. In the present study, we show that the use of an anti-CEA antibody conjugated to an 800nm NIR fluorescent dye can selectively label pancreatic cancer in both pancreatic cancer cell lines and patient derived xenograft mouse models (PDOX). Materials/Methods: BxPC3-GFP cells or pancreatic cancer tissue specimens were grafted into flanks of nude mice. Both types of tumors were allowed to grow until 1 cm. Fragments tumors (2 mm3) were grafted onto the pancreatic tail of recipient mice to create PDOX models. After tumors reached 7-10mm in size, 75 ug of Anti-CEA-800 dye was injected into the tail-vein (LI-COR 800 CW IRDye®, Lincoln, NE). Mice were imaged via the Maestro CRI imaging system (Perkin Elmer, Waltham, MA) 24 hours after injection. Results: Images obtained after fluorescent antibody injection showed that anti-CEA-800 specifically labeled both the cell-line-derived tumors and patient-derived tumors with an adequate tumor to background contrast at 24 hours. The dye co-localized with GFP in tagged tumor cells lines. GFP fluorescence had poor penetration if covered by overlying tissue, which was not an issue with the anti-CEA-800 dye. The dye allowed clear identification of the tumor and tumor margin. In the BxPC-3 based tumor, the dye allowed identification of a small 1mm satellite lesion in the spleen that would otherwise have been missed. Conclusions: Fluorescent humanized anti-CEA-800 antibody specifically labeled orthotopically implanted pancreatic cancer xenografts from cell lines and patient derived tissue. Tumor-specific fluorescence imaging clearly identified the primary tumor and satellite lesion. The 800nm wavelength allowed deeper tissue penetration, particularly in areas of tumor covered by normal pancreatic parenchyma. Humanized anti-CEA antibodies conjugated to a radio-labeling agent and LI-COR 800 dye is already in Phase I/II trials. Humanized anti-CEA conjugated with the IR-800 dye is a promising agent for future clinical FGS applications. Citation Format: Thinzar Lwin, Takashi Murakami, Robert M. Hoffman, Paul J. Yazaki, Michael Bouvet. Tumor-specific antibody labeling of pancreatic cancer in a patient-derived orthotopic xenograft (PDOX) mouse model using a fluorescent humanized anti-CEA antibody [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3729. doi:10.1158/1538-7445.AM2017-3729

Synthesis, characterization and liver targeting evaluation of self-assembled hyaluronic acid nanoparticles functionalized with glycyrrhetinic acid

Recently, polymeric materials with multiple functions have drawn great attention as the carrier for drug delivery system design. In this study, a series of multifunctional drug delivery carriers, hyaluronic acid (HA)-glycyrrhetinic acid (GA) succinate (HSG) copolymers were synthesized via hydroxyl group modification of hyaluronic acid. It was shown that the HSG nanoparticles had sub-spherical shape, and the particle size was in the range of 152.6–260.7nm depending on GA graft ratio. HSG nanoparticles presented good short term and dilution stability. MTT assay demonstrated all the copolymers presented no significant cytotoxicity. In vivo imaging analysis suggested HSG nanoparticles had superior liver targeting efficiency and the liver targeting capacity was GA graft ratio dependent. The accumulation of DiR (a lipophilic, NIR fluorescent cyanine dye)-loaded HSG-6, HSG-12, and HSG-20 nanoparticles in liver was 1.8-, 2.1-, and 2.9-fold higher than that of free DiR. The binding site of GA on HA may influence liver targeting efficiency. These results indicated that HSG copolymers based nanoparticles are potential drug carrier for improved liver targeting.

Photoswitching Near-Infrared Fluorescence from Polymer Nanoparticles Catapults Signals over the Region of Noises and Interferences for Enhanced Sensitivity.

As a very sensitive technique, photoswitchable fluorescence not only gains ultrasensitivity but also imparts many novel and unexpected applications. Applications of near-infrared (NIR) fluorescence have demonstrated low background noises, high tissue-penetrating ability, and an ability to reduce photodamage to live cells. Because of these desired features, NIR-fluorescent dyes have been the premium among fluorescent dyes, and probes with photoswitchable NIR fluorescence are even more desirable for enhanced signal quality in the emerging optical imaging modalities but rarely used because they are extremely challenging to design and construct. Using a spiropyran derivative functioning as both a photoswitch and a fluorophore to launch its periodically modulated red fluorescence excitation energy into a NIR acceptor, we fabricated core-shell polymer nanoparticles exhibiting a photoswitchable fluorescence signal within the biological window (∼700-1000 nm) with a peak maximum of 776 nm. Live cells constantly synthesize new molecules, including fluorescent molecules, and also endocytose exogenous particles, including fluorescent particles. Upon excitation at different wavelengths, these fluorescent species bring about background noises and interferences covering nearly the whole visible region and therefore render many intracellular targets unaddressable. The oscillating NIR fluorescence signal with an on/off ratio of up to 67 that the polymer nanoparticles display is beyond the typical background noises and interferences, thus producing superior sharpness, reliability, and signal-to-noise ratios in cellular imaging. Taking these salient features, we anticipate that these types of nanoparticles will be useful for in vivo imaging of biological tissue and other complex specimens, where two-photon activation and excitation are used in combination with NIR-fluorescence photoswitching.