Government healthcare organizations are facing big challenges as a result of increasing burden of diseases. Since the resources are limited, by restructuring the organizational infrastructure a better use of available resources is possible, which will improve the quality of services. The 5S approach is an effective tool of lean management principles which reduces waste through a better organized workplace, leading to improved quality service. The objectives of this study were to analyse the perceptions of healthcare workers about 5S methodology and examine the feasibility and effectiveness of implementing 5S in a clinical biochemistry laboratory. This interventional study was conducted in three phases. First phase was about advocacy and training of trainers. The second phase included a implementation of 5S and practicing it under supervision. The third phase had review meeting, and filling of the feedback questionnaire. The study revealed that the laboratory has become a more organised and safe place to work. It has helped in developing a positive attitude in the staff towards their responsibilities and inspired others to follow 5S methodology. Implementation of 5S required commitment of top management and total involvement of staff at all levels.