Tailor-made treatment for lymphoma has been proposed, based on the information on prognostic on predictors or on molecular targets. This recent evolutions necessitated the collection of sufficient tissue samples for their preoperative evaluation. Since needle biopsy or needle aspiration cytology may not provide enough tissue for histo-pathologic diagnosis, incisional and excisional biopsies are commonly performed. In this study, we applied a handheld vacuum-assisted biopsy (VAB) system, as an alternative to the conventional incisional and excisional biopsies, to evaluate its feasibility in the biopsy -based diagnosis of malignant lymphoma. The subjects were nine patients with clinically suspected malignant lymphoma. All of them were correctly diagnosed with a diagnostic accuracy of 100%, and lymphoma sub-types could be defined according to the New WHO classification. There was no complication associated with the procedure. Lymph node biopsy for diagnosis of malignant lymphoma using this minimally invasive VAB technique allows a simple and safe collection of sufficient samples with good-quality, and promises to contribute to a precise molecular-based diagnosis.