The purpose of this study was to quantify biomechanical and cardiovascular exposure while making beds with and without interventions (mattress lift tool and fitted sheet). Sixteen female hotel room cleaners participated in this multifactorial (tool and sheet) laboratory study of crossover design. Exertion in the upper extremity (<2) and back (<3) was consistently lower when using the tool and fitted sheet (p < 0.05). The average number of lifts per bed was reduced by 48% with an 18 s increase in cycle time per bed. Peak forearm flexor activity was significantly lower when using a tool(p < 0.05). Spinal lateral plane range of motion (p < 0.02) and maximum twisting velocity (p < 0.03) were lowest using the tool and fitted sheet together. Interventions such as a mattress lift tool used with a fitted sheet reduced the number of mattress lifts and lowered perceived exertion among hotel room cleaners while making beds.