celltrace far red, cfse green fluorescence dye  (Thermo Fisher)

Journal: bioRxiv

Article Title: Induction of senescence renders cancer cells highly immunogenic

doi: 10.1101/2022.06.05.494912

Figure Lengend Snippet: A. Levels of extracellular ATP in the conditioned media (CM) of 10 6 untreated B16F10 cells, dying by immunogenic cell death induced by a high dose of doxorubicin (ICD-B16F10) and senescent B16F10 induced by a low dose of doxorubicin (senB16F10). n =2 independent experiments. **p<0.01 *p<0.05; one-way ANOVA test compared to untreated B16F10. B. Immunoblot detection of CALR in the CM of 10 6 untreated B16F10, ICD-B16F10 and senB16F10. Representative image (left panel) and quantification (right panel) of n =3 independent experiments are shown. *p<0.05; one-way ANOVA test compared to untreated B16F10. C. Flow cytometry analysis of uptake of CFSE (cytosolic dye) or WGA-Alexa647 (membrane dye) by BMDCs from labeled untreated B16F10, ICD-B16F10 or senB10F10. Quantification after subtraction of autofluorescence from unstained BMDC of n =3 independent experiments. ***p< 0.001; **p<0.01 ; *p<0.05, one-way ANOVA test. D. Immunochemistry staining of CD11b + cells (purple) in skin sections of animals 7 days after subcutaneous injection of ICD or senB16F10. Representative images selected by a histopathologist of n =5 animals per group are shown. Note that the brown pigmentation is due to the melanine. Scale bars for each images are shown (100µm). E. In vivo imaging detection of luciferase–expressing B16F10 treated with vehicle ( n =6), ICD-B16F10 ( n =7) or senB16F10 ( n =7) at different time points after subcutaneous injection (as indicted). Representative images (left panels) and quantification (right panel) are shown. *p<0.05; Two-way ANOVA test compared to vehicle-treated group.

Article Snippet: Untreated, ICD, and senescent cancer cells were stained using the fluorescent cytosolic-dye CellTrace CFSE (Thermofisher) or the fluorescent membrane-dye wheat germ agglutinin WGA-Alexa Fluor 647 (Thermofisher).

Techniques: Western Blot, Flow Cytometry, Membrane, Labeling, Staining, Injection, In Vivo Imaging, Luciferase, Expressing

Journal: Frontiers in Microbiology

Article Title: Phenotype-Independent Isolation of Interspecies Saccharomyces Hybrids by Dual-Dye Fluorescent Staining and Fluorescence-Activated Cell Sorting

doi: 10.3389/fmicb.2019.00871

Figure Lengend Snippet: Intra-species mating of S. cerevisiae strains CEN.PK113-5A ( MATa URA3 his3 -Δ 1 leu2-3,112 trp1-289 ) and IMK439 ( MATα HIS3 TRP1 LEU2 ura3 Δ ::KanMX ). (A) Fluorescence contour plots of unstained CEN.PK113-5A, CEN.PK113-5A stained with CellTrace TM CFSE, IMK439 stained with CellTrace TM Violet, and of the mating culture after 18, 24, and 42 h. The indicated gated areas were used for sorting cells, event rates of each gate are indicated as a percentage of total cell counts. (B) Microscope image (400×) of zygotes sorted from the double-stained population (C+V+) after 42 h of mating. (C) Percentage of cells able to grow on synthetic medium without auxotrophy-complementing supplements in different populations sorted by FACS, as indicated in (A) .

Article Snippet: For staining, CellTrace TM Violet, CellTrace TM CFSE and CellTrace TM Far Red fluorescent dyes (Thermo Fisher Scientific, Waltham, MA, United States) were prepared according to the manufacturers’ recommendations.

Techniques: Fluorescence, Staining, Microscopy