2D Toxicity MTS Assay

Basic information

• 2D experiment in 96-well plate
• Cell line: PANC-1 5000 cells/well and 7500 cells/well; P34; viability 82 %
• Nanoparticles: GGAG@SiO₂, GGAG@SiO₂-RB
• Concentration: 0.0, 0.05, 0.1, 1 mg/mL
• Other conditions: total killing with 10% DMSO and blank without cells just MTS solution
• Incubation: 22 h

Experiment procedure

1. 96-well plate, seeding 5000 cells/well in 100µL culture medium (rows A-D),
   seeding 7500 cells/well in 100µL culture medium (row E-H), 24 hours grow
2. Removing medium
3. Adding 100µL nanoparticle’s suspension
4. 22hours incubation cells with nanoparticles
5. Rinsing cells with PBS
6. Absorbance measurement at 485nm
7. Adding 80µL media + 20µL CellTiter Aqueous One Solution Cell Proliferation Assay
8. Absorbance measurement at 485nm
9. 1 hour incubation
10. Absorbance measurement at 485nm
11. 1 hour incubation
12. Absorbance measurement at 485nm
13. 30 minutes incubation
14. Absorbance measurement at 485nm
15. Putting 80uL of the viability solution to new 96-well plate for measurement without cells
16. Absorbance measurement at 485nm

Data processing

• count average of the blank wells (MTS solution without cells)
• subtract the average of the blank from each well
• count average of control wells (cells without nano) = 100 % viability
• count % viability for each well
• count average % viability for each condition

Data

Measurement without nanoparticles after 2.5h incubation