Journal: bioRxiv

Article Title: Inhibitory synaptic vesicles have unique dynamics and exocytosis properties

doi: 10.1101/2020.09.21.289314

Figure Lengend Snippet: (A) Schematic diagram depicting a streptavidin-conjugated quantum dot (QD) conjugated to biotinylated antibodies against the luminal domain of VGAT. (B) Colocalization of VGAT-QD‒loaded inhibitory vesicles (green) and CypHer5E-VGAT‒labeled presynaptic boutons (red) in cultured hippocampal neurons. Scale bar: 1 µm. (C) Three-dimensional trajectory of a VGAT-QD‒loaded inhibitory vesicle overlaid on the x - y plane of a CypHer5E-VGAT‒ labeled presynaptic bouton. The color bar represents elapsed time; electrical stimulation (10 Hz) started at 20 s, and the vesicle underwent exocytosis at 32.0 s. (D) Fluorescence images of the VGAT-QD‒loaded vesicle shown in panel C taken at the indicated times. Scale bar: 0.5 µm. (E) Three-dimensional position, radial distance from the momentary position to the fusion site (R), and fluorescence intensity (F) of the VGAT-QD‒loaded vesicle shown in panel C. Note the photoblinking events (e.g., at approximately 8 s, 13 s and 15 s), confirming the presence of one QD inside the vesicle. Electrical stimuli (10 Hz) were applied for 120 s starting at 20 s (green horizontal bar).

Article Snippet: The biotinylated monoclonal mouse anti-Syt1 antibody (105 311BT, Synaptic Systems) or the biotinylated anti-VGAT antibody (131 103CpH, Synaptic Systems) was conjugated to streptavidin-conjugated quantum dots (cat. A10196, Thermo Fisher Scientific), and vesicles were loaded as described previously ( ).

Techniques: Cell Culture, Labeling, Fluorescence

Journal: iScience

Article Title: Preoperative immune checkpoint inhibition and cryoablation in early-stage breast cancer

doi: 10.1016/j.isci.2024.108880

Figure Lengend Snippet:

Article Snippet: Briefly, one million PBMCs and TILs were washed with 2 mL FACS buffer (PBS [phosphate-buffered saline] containing bovine 1% serum albumin and 0.05 mM EDTA), resuspended in 50 μL FACS buffer and stained with a fixable Aqua viability dye (Invitrogen) and a cocktail of antibodies to the following surface markers: CD8-Qdot 605 (Invitrogen, 3B5), CD4-Qdot 655 (Invitrogen, S3.5), PD-1-PE (BD, MIH4), LAG-3-FITC (Enzo Life Sciences, 17B4), ICOS-PE-Cy7 (eBioscience, ISA-3), TIM-3-APC (R&D Systems, 344823).

Techniques: Recombinant, Saline, Staining, Extraction, Software, Flow Cytometry