Combined device of microporous filter membrane and immunofluorescence for detection and characterization of circulating tumor cells of musculoskeletal sarcomas

• The early detection and evaluation of musculoskeletal sarcoma patients with minimal invasion are still yet to be investigated for their lack of classical biomarkers. • Filtration strategy of Microporous Filter Membrane (MPMF) could efficiently enrich sarcoma cells in blood and functionally preserve cellular activity and feasibility. • The combination of MPMF and immunofluorescence (MPMF-IF) achieved promising results in analyzing patients’ general tumor burden. Musculoskeletal sarcomas are rare neoplasms originating from mesenchymal cells and are generally malignant. The presence and persistence of circulating tumor cells (CTCs) are increasingly investigated for their potential significance in tumor evaluation. Currently, various methods have been reported to efficiently extract and examine CTCs in patients’ circulation. However, sarcomas lack classical biomarkers and the ultimate method which could both realize efficiency and activity preservation in sarcomas is still yet to be investigated. Here, we designed a combined strategy of Microporous Filter Membrane and Immunofluorescence (MPMF-IF) for both detection and characterization of CTCs in patients with sarcomas. We conducted numerical experiments to evaluate capture efficiency and purity of the present method through testing sarcoma cell lines and blood samples. Subsequently, the characteristics of the strategy were further investigated in vivo. Also, the device’s performance was validated in patients of different clinical stages. Satisfying results of high capture efficiency, purity and specificity at the same time preserving cellular activity were confirmed in experiments in vitro. Experiments in vivo certified that the device could functionally analyze tumor burden. In clinical samples, the significant difference was detected between metastatic patients and non-metastatic patients and the results illustrated that CTC counts by MPMF-IF could predict the prognosis of patients. Our MPMF-IF method for the detection and characterization of CTCs enables satisfying accuracy and efficiency. A great potential could be expected in further applying MPMF-IF to predict the prognosis and evaluate the therapeutic effect for patients with musculoskeletal sarcomas.