Journal: Drug Delivery

Article Title: Heparan sulfate targeting strategy for enhancing liposomal drug accumulation and facilitating deep distribution in tumors

doi: 10.1080/10717544.2020.1745326

Figure Lengend Snippet: HS-dependent cellular uptake of GBP-modified pegylated liposomes. (A) The A549 cells were incubated with DiI-labeled liposomes (L-DIL) and GBP-modified L-DIL (L-DIL-GBP) at 37 °C for 4 h and were monitored by confocal microscopy. (B) The cells were incubated with L-DOX and L-DOX-GBP at 37 °C for 24 h, and cellular uptake and drug release were assessed by flow cytometry. (C) The cells were treated for 24 h with L-DOX-GBP (H) in the presence of the indicated concentrations of GAGs, including high-molecular-weight heparin (HMWH), low-molecular-weight heparin (LMWH), chondroitin sulfate type B (CS), and hyaluronic acid (HA), and the cellular uptake of doxorubicin was analyzed. The cellular uptake of the L-DOX treatment group was used to normalize that of the L-DOX-GBP(H) group upon GAG competition. (D) The cells were separately treated with 10 μM L-DOX and L-DOX-GBP at 37 °C for 24 h, and cellular uptake and drug release were assessed by fluorescence microscopy. Magnification: 40×; scale bar: 50 μm. The data are the mean ± SD, averaged from three separate experiments. * p < .05; ** p < .01, two-tailed Student’s t -test.

Article Snippet: Each image was measured by CLSM Z-stack scanning (pinhole: 1.7 μm; Z interval: 1.0 μm between consecutive slides) using a JelloX Biotech system.

Techniques: Modification, Liposomes, Incubation, Labeling, Confocal Microscopy, Flow Cytometry, High Molecular Weight, Molecular Weight, Fluorescence, Microscopy, Two Tailed Test

Journal: Drug Delivery

Article Title: Heparan sulfate targeting strategy for enhancing liposomal drug accumulation and facilitating deep distribution in tumors

doi: 10.1080/10717544.2020.1745326

Figure Lengend Snippet: Drug penetration activity of L-DIL-GBP in spheroids. Heterospheroids composed of A549 and NIH-3T3 cells were incubated with different formulations of L-DIL for 4 h. (A) Penetration capacity was measured by CLSM Z-stack scanning with pinhole: 1.7 μm; Z interval: 1.0 μm between consecutive slides. Nuclei stained by SYTO16; DiI (red). Magnification: 20×; scale bar: 100 μm. (B) Three-dimensional images were reconstructed to illustrate L-DIL or L-DIL-GBP(H) penetration into the heterospheroids. DiI signal (green); nucleus (red). (C) Quantitative analysis between the mean intensity of the Dil signal and the distance to the center of the spheroid. The data are the mean ± SD, averaged from three separate experiments. *, **, and *** indicate p < .05, p < .01 and p < .001 under the two-tailed t -test, respectively.

Article Snippet: Each image was measured by CLSM Z-stack scanning (pinhole: 1.7 μm; Z interval: 1.0 μm between consecutive slides) using a JelloX Biotech system.

Techniques: Activity Assay, Incubation, Staining, Two Tailed Test

Journal: Frontiers in Molecular Neuroscience

Article Title: Deletion of Specific Sphingolipids in Distinct Neurons Improves Spatial Memory in a Mouse Model of Alzheimer’s Disease

doi: 10.3389/fnmol.2018.00206

Figure Lengend Snippet: 5xFAD-Thy1-Cre mice are protected from spine loss. (A) ISH confirms that non-tamoxifen-induced Ugcg f/f//Thy1-CreERT2/EYFP mice (n.i. control) and non-induced 5xFAD /Ugcg f/f//Thy1-CreERT2/EYFP mice (n.i. 5xFAD) express GCS in EYFP-fluorescent neurons (scale bar = 10 μm). The original brown ISH dots have been converted to red fluorescence, as described in Materials and Methods section. For comparison, the original images are depicted in Supplementary Figure . Non-induced mice received solvent injections without tamoxifen. (B) The morphology of EYFP-fluorescent dendrites in the hippocampal dentate gyrus (between lateral 0.96 and 1.2 mm (sagittal)) was analyzed by confocal microscopy and subsequent z-stack reconstruction. A loss of spines is observed in n.i. 5xFAD mice, when compared to n.i. control mice. Intriguingly, spine loss is not observed in 5xFAD-Thy1-Cre mice ( n = 88 (n.i. control); 122 (n.i. 5xFAD); 156 (Thy1-Cre); 227 (5xFAD-Thy1-Cre) dendritic ROIs from n = 3 mice per group; scale bar = 5 μm). Means ± SEM. Statistical analysis for all four groups was performed by one-way ANOVA with Tukey’s test for multiple comparison (95% confidence interval).

Article Snippet: GFAP-stained astrocytes in the hippocampal dentate gyrus (DG) were analyzed by z-stack fluorescence microscopy (Keyence).

Techniques: Control, Fluorescence, Comparison, Solvent, Confocal Microscopy

Journal: Proceedings of SPIE--the International Society for Optical Engineering

Article Title: A space- and time-resolved single photon counting detector for fluorescence microscopy and spectroscopy

doi: 10.1117/12.646482

Figure Lengend Snippet: Principle of operation of the H33D detector. Pulsed laser excitation is used to excite the fluorescence of a sample. A fluorescence photon emitted a few ns after the excitation pulse is collected by the imaging optics and interacts with the photocathode (PC), creating a photoelectron with a wavelength-dependent probability QE. The photoelectron is amplified ~107 times by an apposed MCP stack, generating an electron cloud (cone shape). The measure of the delay between the charge pulse at the back of the MCP and the laser pulse (nanotime τ) is performed with a TDC. A cross-delay line anode (distance to MCP: ~6 mm) collects the charges at both ends of each line, and a timing electronics module converts the differences in arrival time into position information (X, Y). A laser pulse counter built in the readout electronics provides a 4th coordinate, the macrotime T (not shown on this diagram). δ: fixed time delay. V: velocity factor proportional to the actual anode signal propagation velocity.

Article Snippet: A S20 multi-alkali photocathode was deposited on a fused silica window and proximity focused on a MCP stack (Z triplet stack, Photonis-DEP, Brive, FR, EU).

Techniques: Fluorescence, Imaging, Amplification