2D Toxicity MTS Assay

Basic information

• 2D experiment in 96-well plate
• Cell line: MIA PaCa-2 P21; viability 32 %, PANC-1 P33; viability 72 %, 7500 cells/well
• Nanoparticles: GGAG@SiO₂, GGAG@SiO₂-RB
• Concentration: 0.0, 0.05, 0.1, 1 mg/mL
• Other conditions: total killing with 10% DMSO
• For new plate with MTS I put MTS solution which was not placed in incubator, just in falcon
• Incubation: 24 h

Experiment procedure

1. 96-well plate, seeding 7500 cells/well in 50µL culture medium, 24 hours grow
2. Removing medium with aspirator
3. Adding 100µL nanoparticle’s suspension
4. 24hours incubation cells with nanoparticles
5. Rinsing cells with PBS
6. Adding 80µL media + 20µL CellTiter Aqueous One Solution Cell Proliferation Assay
7. Absorbance measurement at 485nm
8. 2 hour incubation
9. Absorbance measurement at 485nm
10. Putting 80uL of the viability solution to new 96-well plate for measurement without cells
    not transfer last “total killing” column and put 80µL of MTS solution from the falcon
11. removing the rest of MTS solution from the first plate
12. measuring absorbance 485 nm new plate - MTS solution and viability solution
13. measuring absorbance 485 nm firs plate - only cells with nanoparticles

Data processing

• count average of the TK wells - blank
• subtract the average of the TK 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
• for plate without cells: count average of the MTS solution as blank

Data

Measurement without nanoparticles after 2h incubation