2D Toxicity MTS Assay MIA PaCa-2

Basic information

• 2D experiment in 96-well plate
• cell line: MIA PaCa-2 7500 cells/well; P38; viability 76 %
• nanoparticles: GGAG:Ce@SiO₂, GGAG:Ce@SiO₂-RB (new batch with RB, less pink colour)
• concentration: 0.0; 0.05; 0.5; 5.0; 50; 500; 1000 µg/mL
• other conditions:
  • total killing (TK) with 10% DMSO (6 wells)
  • blank MTS solution (6 wells)
  • covering the plate with aluminum foil all the time; inclusive of incubation
• growing time approx 24h
• incubation time with NPs 24h
• MTS incubation 2h
• R2 second replication

Experiment procedure

• 96-well plate, seeding 7500 cells/well in 100 µL culture medium
• Growing from: 20.12. 11:05; about 24h
• Removing medium
• Absorbance measurement at 485 nm (cells without any nano or medium)
• Adding 100 µL nanoparticle’s suspension (suspension prepared before the adding),
  nanoparticles mixed by vortex before pipetting into the vials
• Covering with aluminum foil and putting to incubator
• Incubation from: 21.12. 12:30; about 24h
• Rinsing from: 22.12. 12:50
• Rinsing cells with PBS (takes about 0.6h till starting incubation MTS)
• Absorbance measurement at 485 nm
• Adding 80 µL of medium + 20 µL CellTiter Aqueous One Solution Cell Proliferation Assay
• 2 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

Viability table for all wells