such as WHO hemoglobin color scale, Hemocue, NBM-200, and Pronto-7, rely on testing the level of Hb in whole blood. [4,5] For timely and efficient treatment of anemia, beside Hb detection, it is important to monitor other related biomarkers to help identify specific disease etiology. It appears that nutritional deficiencies (such as iron deficiency, folic acid (FA) deficiency, and vitamin B 12 (VB 12 ) deficiency) in underserved communities play a major role in anemia prevalence. [6,7] The deficiency of iron, FA, and VB 12 in pregnant women may affect DNA synthesis and seriously endanger the life safety of both pregnant women and fetuses. Therefore, it is necessary to detect Hb and other anemia biomarkers to treat anemia promptly and properly.Simultaneously quantifying the above biomarkers is challenging because they are distributed in different environments with different concentration ranges. Besides, these biomarkers are detected with different principles that are difficult to be integrated into a single device. Hb is a protein found in red blood cells, whose normal level is 11-16 g dL −1 . [8,9] Other biomarkers, Ferritin (a typical biomarker This work reports a portable, origami-type paper device with a plasmonic fluor-labeled microneedle sensing module for the multiplexed quantification of anemia biomarkers in whole blood. Sequential steps, including serum separation, target enrichment, and multiplexed readout by a gel imager, are rapidly accomplished with the flexible and highly integrated device. The microneedle array enabled efficient sampling of trace targets from ng mL −1 to pg mL −1 level. Combined with the plasmonic fluor label, the signal is improved by ≈7.6 folds compared with the flat substrate-based assay. The device is applied to simultaneously quantify hemoglobin (Hb), ferritin, folic acid (FA), and vitamin B 12 (VB 12 ), which are four anemia biomarkers distributed in different environments with different concentration ranges. Featured by the small sample volume (150 µL), short assay time (20 min), low cost (2 $), robust stability, and user-friendliness, the device is promising for the rapid and accurate diagnosis of anemia in real practice.