This  study  aimed  to  develop  an  optical  stiffness  calibration  system  using  three methods. The used methods were Boltzmann statistics (BS), equipartition theorem (ET) and power  spectrum density (PSD) analysis. This study consisted of two phases; system development phase and  testing phase. System development phase involved hardware and software components. Testing phase  involved actual experimental optical stiffness calibration. The finding of the study was a  calibration system consisting hardware and software parts. The hardware part was an optical  tweezers that was equipped with quadrant photodiode (QPD) and piezostage. The software part was a  custom made software which was constructed on LabVIEW 2012 platform, namely OSCal. This study  showed the conversion factor (used in ET and BS methods) and optical stiffness depends on trapping  laser power. In conclusion, this study successfully developed the aimed system with comparable  optical sti.....

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