Journal: Science and Technology of Advanced Materials

Article Title: Quantitative evaluation of malignant gliomas damage induced by photoactivation of IR700 dye

doi: 10.1080/14686996.2016.1205936

Figure Lengend Snippet: The evaluation of damage and cell death in U87MG and BT142 cells; (a) and (b), the nucleus in the cells stained by EthD-1 at 20 m after photoactivation of IR700. Inset images are phase-contrast images of the corresponding cells; (c) shows the fluorescence intensity of EthD-1 after photoactivation relative to that before photoactivation. The numbers represent the data from six cells; (d) and (e) show fluorescence images of Annexin-V 20 m after photoactivation with loading 40 nM of IR700-CD133 in cells. Scale bar: 10 μm.

Article Snippet: The Annexin-V-Cy5 Apoptosis Detection Kit was purchased from BioVision, Inc. Ethidium homodimer-1 (EthD-1), paraformaldehyde, sodium dihydrogen phosphate dihydrate and sodium dihydrogen phosphate were purchased from Wako Pure Chemicals.

Techniques: Staining, Fluorescence

Journal: Materials Today Bio

Article Title: Viridibacillus culture derived silver nanoparticles exert potent anticancer action in 2D and 3D models of lung cancer via mitochondrial depolarization-mediated apoptosis

doi: 10.1016/j.mtbio.2024.100997

Figure Lengend Snippet: Effect of V-AgNPs on in vitro 3D tumor progression model and 3D tumor size model. (A) Schematic and description show establishment of in vitro tumor progression model in which the A549 cells were seeded (day 0) and on the same day V-AgNPs were added at relatively safer concentration (LD25-3D). The spheroid growth was monitored for the shape and live-dead cell fractions till day 5. Untreated A549 cells i.e. cells forming spheroids in absence of V-AgNPs served as control for this experiment. (B) Representative images of spheroid forming cells (either in absence or presence of V-AgNPs) stained with hoechst and ethidium homodimer II. For each group (control and V-AgNPs treated) total 8 spheroids were imaged per day ( n = 8). Images for all replicates for all days have been given in supporting information, . Images for only 3 replicates are given here. (Scale bar in the images: 200 μm) (C) The graph shows the dead cell fraction (ratio of red fluorescence to blue fluorescence) for untreated and V-AgNPs treated spheroids on each day and each point represents mean fraction ± std. error for 8 replicates ( n = 8). The line has been drawn using spline fitting function to represent the mean trend of rise of dead cell fraction. (D) Schematic and description show establishment of in vitro tumor size model in which the A549 cells were seeded (day 0) and grown till day 5. Then 5-days old spheroids were treated with 2–8 μg/ml concentration of V-AgNPs (the highest dose here is LD25-3D). The diameters of spheroids were measured before (Day 5) and after the treatment (Day 8). Untreated A549 spheroids served as control for this experiment. (E) Representative brightfield images for untreated and 8 μg/ml V-AgNPs treated cells on day 5 and day 8. The red and yellow dotted lines represent the diameters of spheroid before and after the V-AgNPs treatment, respectively (Scale bar in the images: 500 μm). The inset in the images shows differences in diameter before and after the treatment. The V-AgNPs can be seen around the spheroids as shown by blue arrow in 8th day V-AgNPs treatment image. The third row shows zoomed-in image of untreated and V-AgNPs treated spheroids. The brightfield images for all treatment groups (2–8 μg/ml) are given in supporting information, . (F) The bar graph shows the inter-day comparison of diameter of untreated (control) and 2–8 μg/ml V-AgNPs treated spheroids and each bar represents mean diameter ± std. error for 25 replicates ( n = 25). The intra-day comparison is given in supporting information, . The asterisks in the bar graph represent significantly different observations (One-way ANOVA test: * p-value<0.05, **p-value<0.01, *** p-value<0.001, **** p-value<0.0001). (Abbreviations – V-AgNPs: Viridibacilli derived silver nanoparticles, t: time, d: diameter, R/B ratio: ratio of Red and Blue fluorescence intensities) (Schematics were created with BioRender.com ). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Epidermal growth factor (EGF), CellEventTM Caspase-3/7 Green ReadyProbesTM, hoechst, ethidium homodimer II, 4′,6-diamidino-2-phenylindole (DAPI) were purchased from ThermoFisher Scientific, Denmark.

Techniques: In Vitro, Concentration Assay, Control, Staining, Fluorescence, Comparison, Derivative Assay

Journal: The British Journal of Ophthalmology

Article Title: Global cell-by-cell evaluation of endothelial viability after two methods of graft preparation in Descemet membrane endothelial keratoplasty

doi: 10.1136/bjophthalmol-2015-307534

Figure Lengend Snippet: (A) TRITC/ethidium channel image showing manual demarcation of graft edge using the polygon selection toll in Image J (yellow outline). (B) Yellow line defined in the TRITC/ethidium image is copied and applied to the FITC/calcein channel image. The edge of calcein-positive area is defined in the same way as the graft edge (blue outline). The difference in the areas of the yellow and blue selections was defined as the trephination damage area. (C) The FITC/calcein channel image is thresholded and segmented. The area of living cells (red area) is used to create a selection mask (green lines). (D) Ultraviolet/Hoechst channel images were colour inverted and the viable graft area selection applied (green outline). Each nucleus (black dot) within the selection mask is counted. Zoomed in area (top right) showing all nuclei within the mask have been counted, as shown by the presence of a red dot in the centre of nucleus. Nuclei outside the mask are not counted (no red dot in centre of nucleus). (E) TRITC/ethidium channel image is inverted, and the living area selection is applied (green outline). Each nucleus (black dot) within the selection mask is counted. Zoomed in area (top right) showing ethidium-staining nuclei within the mask have been counted, as shown by the presence of a red dot in the centre of nucleus.

Article Snippet: Each sample was covered with 250 μL of BSS containing Hoechst 33342 (10 μM), ethidium homodimer II(4 μM) and calcein-AM (2 μM) (Invitrogen, Carlsbad, California, USA) and incubated at 37°C for 30 min. Hoechst 33342 and ethidium homodimer II are nuclear binding dyes that do not fluoresce until they are bound to nuclear DNA.

Techniques: Selection, Staining