Determining the precise role of molecular factors present in venous leg ulcer exudate will expedite the identification of biomarkers that can optimally guide treatment. However, there is now no standardized approach for collecting, processing and storing wound fluid samples for molecular analyses. This scoping review was conducted to integrate and summarize the multiple types of methods being used currently in studies of venous leg ulcers for collecting, processing and storing wound fluid prior to analysis. PubMed, CINAHL, EMBASE and Scopus databases were searched for eligible studies between 2012 and 2022. Nineteen studies were selected for this scoping review. Five primary methodological categories for wound fluid sampling were identified. The most commonly used collection method involved extracting the fluid from various absorbent materials, and the majority of studies centrifuged wound fluid before storing it at ultra‐low temperatures. This review found the wound fluid sampling methods among the included studies to be heterogeneous. Moreover, the data revealed no definitive patterns. There is a critical need to develop standardized wound fluid sampling methods in research to facilitate accurate comparisons of biomarker data across studies and a more rapid determination of biomarkers that can most effectively guide delivery of tailored venous leg ulcer treatments.