alamar blue assay  (Biotium)

Journal: Journal of Advanced Research

Article Title: Mitochondrial fatty acid oxidation as the target for blocking therapy-resistance and inhibiting tumor recurrence: The proof-of-principle model demonstrated for ovarian cancer cells

doi: 10.1016/j.jare.2025.03.026

Figure Lengend Snippet: Paclitaxel induced transient ovarian cancer persister cells. ( A ) Schematic representation of paclitaxel-tolerant persister cell generation. ( B ) Representative morphological images of parental, persister, regrown and resistant cells captured under a phase contrast microscope. Scale bar, 100 μm. ( C ) OVCAR8 and A2780 cells were treated with 20 nM paclitaxel for 9 days and then withdrawn. The cell viability was evaluated daily using Alamar Blue assay for 40 days and normalized to that on day 1. ( D ) Cell proliferation was measured by CFSE dilution assay. CFSE divides between daughter cells upon cell division and can be tracked by decreasing the fluorescence intensity. ( E ) Parental, persister, and regrown cells derived from OVCAR8 (left) and A2780 (right) cell lines were treated with various concentrations of paclitaxel ranging from 1 to 1 × 10 4 nM for 48 h, and cell viability was assessed using CCK-8 assay. ( F ) Heatmap representing the IC 50 values of paclitaxel at 48 h in parental, persister and regrown cells derived from OVCAR8 (left) and A2780 (right) cell lines. ( G ) Quantification of annexin V-positive apoptotic parental, persister, and regrown cells derived from OVCAR8 (left) and A2780 (right) cell lines after vehicle or paclitaxel treatment. CFSE, carboxyfluorescein diacetate succinimidyl ester. PTX, paclitaxel. IC 50 , half maximal inhibitory concentration. ns, not statistically significant; ** P < 0.01; *** P < 0.001. The values are presented as the means ± SDs.

Article Snippet: Cell proliferation was measured using Alamar Blue (Yeasen, China) assay, carboxyfluorescein diacetate succinimidyl ester (CFSE, MedChemExpress, USA) dilution assay or cell counting kit-8 (CCK-8, Beyotime, China) assay; and cytotoxicity was evaluated by CCK-8 assay or CellTiter-Glo® 2.0 cell viability assay (Promega, USA) according to the manufacturer’s protocol (see “Cell proliferation and cytotoxicity” in the Supplementary Material for additional details).

Techniques: Microscopy, Alamar Blue Assay, Dilution Assay, Fluorescence, Derivative Assay, CCK-8 Assay, Concentration Assay

Journal: HardwareX

Article Title: AnyBio – An easy off-the-shelf masked stereolithography bioprinter conversion combined with radical-scavenging strategies

doi: 10.1016/j.ohx.2025.e00705

Figure Lengend Snippet: Validating Thermal conditions and bioactivity in the presence of Vitamin C. a) Experimental layout for measuring the photothermal effects, b) with the probe reaching into the bioresin. c) The thermal amplitude normalized to the starting temperature of each individual printing run (n = 4) as boxplots at 33 °C and 37 °C environmental temperature (corresponding to 36/40 °C heater setpoint) respectively. More heat build-up was observed at 37 °C (dT = 3.38 °C) than 33 °C (dT = 2.75 °C) ambient temperature, though results were not statistically significant. Significance was tested with Welch’s t -test: p = 0.104; Bayesian posterior 95 % CI = [–0.09, 1.28], see analysis script for quantifications) d) Heat build-up in the bioresin over time (40 layers, 23 mins) depicted as line graphs with bold lines representing the average heating at 33 °C and 37 °C environmental temperature (n = 3). e) Fluorescent images of Live/Dead assays of casted samples with and without Vitamin C and printed samples with Vitamin C (GelMA). Scale bar = 100 µm. f) Quantification of cell viability prior to casting, after casting and after printing. Differences between conditions were statistically not significant according to One-way ANOVA (n = 3). g) Brightfield images of the printed discs including HEK cells over a period of 6 days. Scale bar = 300 µm. h) Alamar Blue readout depicted as fold change metabolic activity and plotted as boxplots (n = 4) over a period of 6 days. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: To assess long-term viability and metabolic activity of printed constructs, perform Alamar Blue (AB) assays using phenol red–free DMEM/F-12 medium (SILAC Advanced DMEM/F-12 Flex, Gibco A24943-01) supplemented with 1 g/L D-(+)-glucose, 1 % GlutaMAX (Gibco, 35050–038), 1 % MEM Non-Essential Amino Acids (Gibco, 11140–035), and 1 % Penicillin–Streptomycin (Gibco, 15140–122).

Techniques: Activity Assay