Uncertainties remain about the overall effect of sit-stand desks for reducing prolonged sitting among office-based workers. This study assessed the feasibility of a randomised controlled trial of the impact of workplace sit-stand desks on overall energy expenditure, sitting time and cardio-metabolic outcomes. It involved four phases: Phase I: online survey; Phase II: workspace auditing; Phase III: randomised intervention (provision of sit-stand desks at work for 3 months); Phase IV: qualitative component. Participants were offıce-based employees of two companies in Cambridge, England. Among Phase I participants interested in the trial, 100 were randomised to Phase II. Of those with workspaces suitable for sit-stand desks, 20 were randomised to Phase III. Those allocated to the intervention completed Phase IV. Outcomes included: trial participation interest, desk-type (full desks/desk mounts) and assessment location (work/laboratory/home) preferences (Phase I); proportion of workspaces permitting sit-stand desk installation (Phase II); energy expenditure, sitting time and cardio-metabolic outcomes (Phase III); study participation experiences (Phase IV). Data were collected between May 2015 and December 2016. Recruitment and trial implementation were feasible: 92% of survey respondents expressed participation interest; 80% of workspaces could accommodate sit-stand desks; assessments were done in workplaces, preferred by 71%. Sit-stand desk provision reduced workplace sitting time by 94 min/day (95% CI 17.7–170.7). Their impact on energy expenditure and cardio-metabolic outcomes is unclear. The results confirm the feasibility of a trial assessing sit-stand desks’ impact on energy expenditure, sitting time and cardio-metabolic outcomes, which should reduce uncertainty concerning the intervention’s potential to reduce the health risks of prolonged sitting.