PEG Coated Biocompatible Cadmium Chalcogenide Quantum Dots for Targeted Imaging of Cancer Cells

Cancer stands as a leading cause of mortality worldwide and diagnostics of cancer still faces drawbacks. Optical imaging of cancer would allow early diagnosis, evaluation of disease progression and therapy efficiency. To that aim, we have developed highly biocompatible PEG functionalized cadmium chalcogenide based three differently luminescent quantum dots (QDs) (CdS, CdSe and CdTe). Folate targeting scheme was utilized for targeting cancer cell line, MCF-7. We demonstrate the biocompatibility, specificity and efficiency of our nanotool in detection of cancer cells sparing normal cell lines with retained fluorescence of functionalized QDs as parental counterpart. This is the first time report of utilizing three differently fluorescent QDs and we have detailed about the internalization of these materials and time dependent saturation of targeting schemes. We present here the success of utilizing our biocompatible imaging tool for early diagnosis of cancer.     

Fluorescent Biological Label for Recognizing Human Ovarian Tumor Cells Based on Fluorescent Nanoparticles

In this paper, we report a method for recognizing human ovarian tumor(HOT) cells using fluorescent biological label based on core-shell nanoparticles. The luminescent nanoparticles were synthesized with a water-in-oil(W/O)micromulsion technique. The fluorescent silica core-shell nanoparticles modified with anti-HER2 antibody using bifunctional cross-linker glutaraldedhyde targeted the corresponding tumor antigen in the cell surface of the SKOV3 ovarian cancer cells. The specific immunoreactivity of antibody-nanoparticles with cells was characterized by laser scanning microscopy (LSM) and scanning electron microscope (SEM). The results showed that the method offered potential advantages of sensitivity and simplicity due to high binding efficiency between nanoparticles and cells and provided an alternative method for the detection of HOT.     

Preparation and Non-Invasive In-Vivo Imaging of Anti-Adhesion Barriers with Fluorescent Polymeric Marks

Comb-like PEMLn polymers with pendent PEG-PLLA side chains were synthesized as tissue anti-adhesion barriers. The comb-like structure improved the flexibility of the films. Fluorescent polymer-biocompatible polymer guest-host materials were printed on the films as marking dots. Without sacrificing rats on different days after surgery, degradation behaviors of the marked films can be investigated non-invasively in the in-vivo imaging system (IVIS) by monitoring the location of fluorescent signals. Degradation properties of PEML1/G26L35 films were adjusted by incorporating G26L35 oligomers. PEML1 and PEML1/G26L35 films were very effective in preventing post-surgical tissue-adhesions. Degradation behaviors of various films observed in the animal study were consistent with those investigated by the in-vivo imaging method. Fluorescent polymer/biocompatible polymer blends were promising candidates for in-vivo imaging applications.     

Accurate Sensitivity of Quantum Dots for Detection of HER2 Expression in Breast Cancer Cells and Tissues

Here we introduce novel optical properties and accurate sensitivity of Quantum dot (QD)-based detection system for tracking the breast cancer marker, HER2. QD525 was used to detect HER2 using home-made HER2-specific monoclonal antibodies in fixed and living HER2⁺ SKBR-3 cell line and breast cancer tissues. Additionally, we compared fluorescence intensity (FI), photostability and staining index (SI) of QD525 signals at different exposure times and two excitation wavelengths with those of the conventional organic dye, FITC. Labeling signals of QD525 in both fixed and living breast cancer cells and tissue preparations were found to be significantly higher than those of FITC at 460–495 nm excitation wavelengths. Interestingly, when excited at 330–385 nm, the superiority of QD525 was more highlighted with at least 4–5 fold higher FI and SI compared to FITC. Moreover, QDs exhibited exceptional photostability during continuous illumination of cancerous cells and tissues, while FITC signal faded very quickly. QDs can be used as sensitive reporters for in situ detection of tumor markers which in turn could be viewed as a novel approach for early detection of cancers. To take comprehensive advantage of QDs, it is necessary that their optimal excitation wavelength is employed.     

Photodynamic Therapy for Cancer Cells Using Metal-Halide Lamps

New metal-halide lamps were developed and their effect on the efficiency of photodynamic therapy (PDT) for cancer cells, murine thymic lymphoma cells (EL-4), was investigated. 5-Aminolevulinic acid-induced protoporphyrin IX was used as a photosensitizer. The metal-halide lamps were made by introducing sodium iodide (Na lamp), lithium iodide (Li lamp), and sodium iodide-lithium iodide mixture (Na-Li lamp) into their discharge tubes. These lamps emitted light in the range of 550 to 750 nm and had specific emission peaks at 580 and 600 nm for the Na lamp, 580, 610, and 680 nm for the Li lamp, and 580, 610, and 675 nm for the Na-Li lamp. Changes in the survival rate of EL-4 with increasing irradiation time indicated that PDT efficiency of the lamps increased in the order Li lamp < Na lamp < Na-Li lamp. We also found that a dark interval during irradiation of the light with the Na-Li lamp enhanced PDT efficiency.     

Advanced Nanotools for Imaging of Solid Tumors and Circulating and Disseminated Cancer Cells

We propose methods for creating spherical gold particles of submicron size and silver rod-like particles with transverse dimensions of ~10 nm and an aspect ratio of 1: 10. Factors determining the frequency of plasmon resonances are considered, reagents are selected, and their ratios for obtaining prolate silver particles are determined. An optimal concentration of the surfactant is determined for creating most elongated silver particles. A shift of the plasmon absorption toward the near-IR range of the spectrum is obtained.     

A Nitronaphthalimide Probe for Fluorescence Imaging of Hypoxia in Cancer Cells

The bioreductive enzymes typically upregulated in hypoxic tumor cells can be targeted for developing diagnostic and drug delivery applications. In this study, a new fluorescent probe 4?(6?nitro?1,3?dioxo?1H?benzo[de]isoquinolin?2(3H)?yl)benzaldehyde (NIB) based on a nitronaphthalimide skeleton that could respond to nitroreductase (NTR) overexpressed in hypoxic tumors is designed and its application in imaging tumor hypoxia is demonstrated. The docking studies revealed favourable interactions of NIB with the binding pocket of NTR-Escherichia coli. NIB, which is synthesized through a simple and single step imidation of 4?nitro?1,8?naphthalic anhydride displayed excellent reducible capacity under hypoxic conditions as evidenced from cyclic voltammetry investigations. The fluorescence measurements confirmed the formation of identical products (NIB-red) during chemical as well as NTR?aided enzymatic reduction in the presence of NADH. The potential fluorescence imaging of hypoxia based on NTR-mediated reduction of NIB is confirmed using in-vitro cell culture experiments using human breast cancer (MCF?7) cells, which displayed a significant change in the fluorescence colour and intensity at low NIB concentration within a short incubation period in hypoxic conditions.

Graphical abstract

     

SiO_2/Ag核壳结构纳米粒子的制备及其表面荧光增强

以罗丹明B掺杂的SiO2球为核,通过化学还原的方法制备了二氧化硅/银核壳结构复合纳米粒子。采用透射电镜(TEM)、紫外-可见-近红外(UV-Vis-NIR)分光光度计和荧光分光光度计对二氧化硅/银核壳结构纳米粒子的表面形貌、表面等离子共振和表面荧光增强特性进行了研究和表征。结果表明,二氧化硅/银核壳结构纳米粒子的表面等离子共振峰具有明显的可调谐性,且其表面荧光增强强烈依赖于银壳层的表面等离子共振,随银壳层厚度的增大而增强。     

A Novel Highly Sensitive and Selective Fluorescent Sensor for Imaging Copper (II) in Living Cells

In this work, we designed and synthesized a novel quinolin-based derivative which exhibited signaling behaviors for Cu²⁺. Upon the addition of Cu²⁺ to the solution of the molecule, it displayed an obvious fluorescence quenching in a linear fashion due to the formation of a 1:1 metal–ligand complex. This fluorescent sensor exhibited a rare sensitivity toward Cu(II) (the level of magnitude could be 6?×?10^?8), a rapid response (<10 s) and also high selectivity toward Cu²⁺ over other metal ions such as Na⁺, K⁺, Ca²⁺, Mg²⁺, Ba²⁺, Fe³⁺, Mn²⁺, Hg²⁺, Pb²⁺, Cd²⁺, Cr³⁺, Co²⁺, Zn²⁺ and Al³⁺. Simultaneously, the cell imaging experiments and filter paper test demonstrated its extensive applicability.     

A Ratiometric and near-Infrared Fluorescent Probe for Imaging Cu<Superscript>2+</Superscript> in Living Cells and Animals

We have rationally constructed a novel ratiometric and near-infrared Cu²⁺ fluorescent probe based on a tricarbocyanine chromophore. The new probe NIR-Cu showed a ratiometric fluorescent response to Cu²⁺ with a large emission wavelength shift (up to 142 nm) in the far-red to near-infrared region. The probe also displayed a large variation in the fluorescence ratio (I₆₃₆/I₇₇₈) to Cu²⁺ species with high sensitivity and selectivity. Additionally, the developed probe NIR-Cu was suitable for fluorescence imaging of Cu²⁺ in living cells and mice.     

Magnetic Relaxometry with an Atomic Magnetometer and SQUID Sensors on Targeted Cancer Cells

Magnetic relaxometry methods have been shown to be very sensitive in detecting cancer cells and other targeted diseases. Superconducting Quantum Interference Device (SQUID) sensors are one of the primary sensor systems used in this methodology because of their high sensitivity with demonstrated capabilities of detecting fewer than 100,000 magnetically-labeled cancer cells. The emerging technology of atomic magnetometers (AM) represents a new detection method for magnetic relaxometry with high sensitivity and without the requirement for cryogens. We report here on a study of magnetic relaxometry using both AM and SQUID sensors to detect cancer cells that are coated with superparamagnetic nanoparticles through antibody targeting. The AM studies conform closely to SQUID sensor results in the measurement of the magnetic decay characteristics following a magnetization pulse. The AM and SQUID sensor data are well described theoretically for superparamagnetic particles bound to cells and the results can be used to determine the number of cells in a cell culture or tumor. The observed fields and magnetic moments of cancer cells are linear with the number of cells over a very large range. The AM sensor demonstrates very high sensitivity for detecting magnetically labeled cells does not require cryogenic cooling and is relatively inexpensive.     

Photodynamic Therapy for Cancer Cells Using a Flash Wave Light Xenon Lamp

We determined photodynamic therapy (PDT) efficacy using a flash wave (FW) and a continuous wave (CW) light, of which the fluence rate was 70 W/cm², for murine thymic lymphoma cells (EL-4) cultivated in vitro. The irradiation frequency and the pulse width of the FW light were in the range of 1–32 Hz and less than one millisecond, respectively. 5-Aminolevulinic acid-induced protoporphyrin IX (ALA-PpIX) was used as a photosensitizer. When EL-4 with ALA administration was irradiated by the light for 4 h (irradiation fluence: 1.0J/cm²), the survival rate of EL-4 by the FW light was lower than that by the CW light, except for the FW light with irradiation frequency of 32 Hz, and decreased gradually with decreasing irradiation frequency. Moreover, the FW light, especially at lower irradiation frequency, was superior to the CW light for the generation of singlet oxygen in an aqueous PpIX solution. Therefore, thehigher PDT efficacy for EL-4 of the FW light would be caused by the greater generation of singlet oxygen in the cells.     

2-NBDG Fluorescence Imaging of Hypermetabolic Circulating Tumor Cells in Mouse Xenograft model of Breast Cancer

Objectives

To determine use of 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D-glucose (2-NBDG) as a tracer for detection of hypermetabolic circulating tumor cells (CTC) by fluorescence imaging.

Procedures

Human breast cancer cells were implanted in the mammary gland fat pad of athymic mice to establish orthotopic human breast cancer xenografts as a mouse model of circulating breast cancer cells. Near-infrared fluorescence imaging of the tumor-bearing mice injected with 2-DeoxyGlucosone 750 (2-DG 750) was conducted to assess glucose metabolism of xenograft tumors. Following incubation with fluorescent 2-NBDG, circulating breast cancer cells in the blood samples collected from the tumor-bearing mice were collected by magnetic separation, followed by fluorescence imaging for 2-NBDG uptake by circulating breast cancer cells, and correlation of the number of hypermetabolic circulating breast cancer cells with tumor size at the time when the blood samples were collected.

Results

Human breast cancer xenograft tumors derived from MDA-MB-231, BT474, or SKBR-3 cells were visualized on near-infrared fluorescence imaging of the tumor-bearing mice injected with 2-DG 750. Hypermetabolic circulating breast cancer cells with increased uptake of fluorescent 2-NBDG were detected in the blood samples from tumor-bearing mice and visualized by fluorescence imaging, but not in the blood samples from normal control mice. The number of hypermetabolic circulating breast cancer cells increased along with growth of xenograft tumors, with the number of hypermetabolic circulating breast cancer cells detected in the mice bearing MDA-MB231 xenografts larger than those in the mice bearing BT474 or SKBR-3 xenograft tumors.

Conclusions

Circulating breast cancer cells with increased uptake of fluorescent 2-NBDG were detected in mice bearing human breast cancer xenograft tumors by fluorescence imaging, suggesting clinical use of 2-NBDG as a tracer for fluorescence imaging of hypermetabolic circulating breast cancer cells.     

A New Pyrazoline-Based Fluorescent Probe for Cu2+ in Live Cells

A new pyrazoline-based probe was synthesized and the structure was determined by using X-ray diffraction analysis. The probe responds to Cu²⁺ in aqueous medium in “turn-off” fluorescent manner with selectivity and sensitivity. Furthermore, the probe could be used for real-time tracking of Cu²⁺ in Hela cells.     

一种可用于细胞内汞离子可视化检测的荧光探针

将硫代罗丹明6G酰肼(R6GS)应用于水溶液和活细胞内汞离子的荧光成像检测.在测试体系中,R6GS本身溶液无色,荧光非常弱;加入汞离子后,溶液立刻变为粉红色且荧光显著增强,而对其他离子则没有响应,表明该探针分子对汞离子有较高的选择性和较高的灵敏性.荧光光谱显示R6GS对0-1.0×10-5mol/L的汞离子有较好线性关系,检出限为6.25×10-7mol/L.最后,该探针被成功用于官颈癌细胞中Hg2+的荧光成像研究.     

Fluorescence Imaging of Stem Cells,Cancer Cells and Semi-Thin Sections of Tissues using Silica-Coated CdSe Quantum Dots

Trioctylphosphine oxide capped cadmium selenide quantum dots, synthesized in organic media were rendered water soluble by silica overcoating. Silanisation was done by a simple reverse microemulsion method using aminopropyl silane as the silica precursor. Further, the strong photoluminescence of the silica-coated CdSe quantum dots has been utilized to visualize rabbit adipose tissue-derived mesenchymal stem cells (RADMSCs) and Daltons lymphoma ascites (DLA) cancerous cells in vitro. Subsequently the in vivo fluorescence behaviours of QDs in the tissues were also demonstrated by intravenous administration of the QDs in Swiss albino mice. The fluorescence microscopic images in the stem cells, cancer cells and semi-thin sections of mice organs proved the strong luminescence property of silica-coated quantum dots under biological systems. These results establish silica-coated CdSe QDs as extremely useful tools for molecular imaging and cell tracking to study the cell division and metastasis of cancer and other diseases.     

A Sandwiched Biological Fluorescent Probe for the Diagnosis of Human Ovarian Tumor Based on TiO2 Nanoparticles

In this paper, we report a novel biological fluorescent probe for the diagnosis of human ovarian tumor based on sandwiched TiO₂ nanoparticles. The fluorescence nanoparticles consist of a fluorescent molecule, tetramethyl rhodamine isothiocyanate (TRITC), sandwiched between titanium dioxide (TiO₂) nanoparticles and nano-gold via reacting with each other. The antibodies HER2, labeled on the surface of the biofluorescence nanoparticles, have granted nanoparticles the privilege of aiming at peculiar tumor antigen. The specificity of antibody-nanoparticles interacting with cells was characterized by Laser Scanning Confocal Microscope. The results showed that these sandwiched nanoparticles were innocuous and stable, and the method offered potential advantages of sensitivity and simplicity due to high combing efficiency between nanoparticles and cells and provided an alternative method for the diagnosis of human ovarian tumor (HOT).     

Preparation and Fluoroimmunoassay Application of New Red-Region Fluorescent Silica Nanoparticles

This is the first report on the preparation and utilization of a novel red-region fluorescent dye (tetracarboxy aluminum phthalocyanine) doped silica nanoparticles. In these nanoparticles, the tetracarboxy aluminum phthalocyanine molecules were covalently bound to silica matrix to protect the dye leaking from nanoparticles in bio-applications. The surface of the nanoparticles was modified by amino groups and easily bioconjugated with goat anti-human IgG antibody. By employing these nanoparticles as fluorescent probe, a sensitive fluoroimmunoassay method has been developed for the determination of trace level of human IgG. The calibration graph for human IgG was linear over the range of 0–500 ng mL⁻¹ with a detection limit of 1.6 ng mL⁻¹. Compared with the corresponding system using free AlC₄Pc as a probe for determining human IgG, the sensitivity of the proposed system was notably increased. The method was applied to the analysis of human IgG in human sera with satisfactory results.