Decalcification is a prerequisite for microscopic examination if any kind of hard tissue received by the histopathology laboratory. The aim of the study was to evaluate various decalcifying chemical agents and to determine the best method for end point determination 40 extracted premolars were decalcified using different decalcifying agents like 10% formal-nitric acid, microwave assisted decalcification using 5% formal-nitric-acid, 10% formal formic acid and 5% formal EDTA solution with pH adjusted to neutral by addition of sodium hydroxide. The end point of decalcification was determined by physical, chemical and radiological methods. After decalcification, the specimens were processed and stained with haematoxylin and eosin. It was found that microwave assisted decalcification with 5% formal nitric acid was the fastest decalcifying agent for human permanent teeth followed by 10% formal nitric acid, 10% formal-formic acid and 5% formal EDTA solution. However the preservation of tissue morphology after decalcification was the best with 5% formal EDTA.