2D Toxicity MTS Assay PANC-1

Basic information

• 2D experiment in 96-well plate
• Cell line: PANC-1 5000 cells/well; P43; viability 42 %
• Nanoparticles: GGAG:Ce@SiO₂, GGAG:Ce@SiO₂-RB
• Concentration: 0.0; 0.05; 0.5; 5.0; 50; 500; 1000 µg/mL
• Other conditions:
  • total killing with 10% DMSO (6 wells)
  • blank without MTS solution (6 wells)
  • covering the plate with aluminum foil all the time (inclusive of incubation)
• Growing time approx. 26h
• Incubation time 23h
• MTS incubation 4h
• R2 second replication

Experiment procedure

• 96-well plate, seeding 5000 cells/well in 100 µL culture medium
• Growing from: 29.11. 13:05; about 26h
• Removing medium
• Absorbance measurement at 485 nm (cells without any nano or medium)
• Adding 100 µL nanoparticle’s suspension (new suspension prepared)
• Covering with aluminum foil and putting to incubator
• Incubation from: 30.11. 16:25; about 23h
• Rinsing cells with PBS
• Absorbance measurement at 485 nm
• Adding 80 µL of medium + 20 µL CellTiter Aqueous One Solution Cell Proliferation Assay
• 2.5 hours incubation
• Absorbance measurement at 485 nm
• 1.5 hours incubation
• Absorbance measurement at 485 nm
• Putting 80 µL of the viability solution to new 96-well plate for measurement without cells
• Removing the rest of viability solution from the previous plate
• Absorbance measurement at 485 nm both plates

Data processing

• count average of the blank wells (MTS solution without cells)
• BLANK CORRECTED: subtract the average of the blank from each well
• count average of control wells (cells without nano) = 100 % viability
• VIABILITY %: count % viability for each well
• count average % viability for each condition
• count standard deviation of the sample (function STDEV.S)

Data

Measurement without nanoparticles