The cell survival rate in the 3D cell constructs was assessed on days 1 and 3 after bio-fabrication and BCG treatment. A fluorescent live/dead staining solution (Thermo Fisher Scientific, Waltham, MA, USA) was used according to the manufacturer’s instructions. Each 3D-cell construct and 2D-cultured cells were washed in Dulbecco’s phosphate buffered saline (DPBS) three times before staining. The DPBS mixture with Calcein-AM (2 µM) and EthD-1 (4 µM) was filtered through a 0.22-mm syringe filter (Sigma, St. Louis, MO, USA). Cell morphologies were observed under a fluorescence microscope (DMI8; Leica, Wetzlar, Germany). Three independent samples were analyzed.
Fluorescent live dead staining solution
Manufactured by Thermo Fisher Scientific
Sourced in United States
Fluorescent live/dead staining solution is a laboratory reagent used to identify and distinguish between live and dead cells in a sample. The solution contains two fluorescent dyes that differentially stain live and dead cells, allowing for their visualization and quantification under a fluorescence microscope or flow cytometer.
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4 protocols using fluorescent live dead staining solution
1
Assessing Cell Viability in 3D Constructs
2
Fluorescent Viability Assessment of 3D Constructs
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The cell survival rate in the 3D cell constructs was assessed at one and three days after fabrication. A fluorescent live/dead staining solution (Thermo Fisher) was used. Each 3D cell construct and the 2D cultured cells were washed in Dulbecco’s PBS (DPBS) three times before staining. The DPBS mixture with Calcein-AM (2 µM) and EthD-1 (4 µM) was filtered through a 0.22-µm syringe filter (Sigma-Aldrich). Cell morphologies were observed under a fluorescence microscope (Leica DMi8, Leica). Three independent samples were analyzed.
3
Assessing Cell Survival in rBCG-sic System
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The cell survival rate in the high-throughput BCOC system was assessed at 24 and 72 hours after rBCG-sic treatment. A fluorescent live/dead staining solution (Thermo Fisher Scientific, Waltham, MA, USA) was used according to the manufacturer’s instructions. Cell morphologies were observed using a fluorescence microscope (DMI8; Leica, Wetzlar, Germany). Three independent samples were analyzed.
4
Assessing 3D Cell Construct Viability
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Cell survival rate in the 3D cell constructs was assessed on days 1, 3, and 5 after biofabrication. A fluorescent live/dead staining solution (Thermofisher, MA, USA) was utilized according to the manufacturer’s instructions. Each 3D cell construct and 2D culture cell sample was washed in PBS three times before staining. The mixture of Calcein-AM (2 μM) and EthD-1 (4 μM) was filtered through a 0.22-mm syringe filter (Sigma, MO, USA). The cell morphologies were observed under fluorescence microscopy (Leica DMI8). Three independent samples were observed.
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