Trauma patients are often transfused allogeneic red blood cells (RBCs) in an effort to augment tissue oxygen delivery. However, the effect of RBC transfusion on microvascular perfusion in this patient population is not well understood. To this end, we investigated the effect of RBC transfusion on sublingual microvascular perfusion in trauma patients. Sublingual microcirculation was imaged at bedside with a sidestream dark-field illumination microscope before and after transfusion of one RBC unit in hemodynamically stable, anemic trauma patients. The perfused proportion of capillaries (PPC) before and after transfusion was determined, and the percent change in capillary perfusion following transfusion (ΔPPC) calculated. Sublingual microcirculation was observed in 30 patients. Mean age was 47 (SD, 21) years, mean Injury Severity Score was 29 (SD, 16), and mean pretransfusion hemoglobin was 7.5 (SD, 0.9) g/dL. No patients had a mean arterial pressure of less than 65 mmHg (mean, 89 [SD, 17] mmHg) or lactate of greater than 2.5 mmol/L (mean, 1.1 [SD, 0.3] mmol/L). Following transfusion, ΔPPC ranged from +68% to -36% and was found to inversely correlate significantly with pretransfusion PPC (Spearman r = -0.63, P = 0.0002). Pretransfusion PPC may be selectively deranged in otherwise stable trauma patients. Patients with relatively altered baseline PPC tend to demonstrate improvement in perfusion following transfusion, whereas those with relatively normal perfusion at baseline tend to demonstrate either no change or, in fact, a decline in PPC. Bedside sublingual imaging may have the potential to detect subtle perfusion defects and ultimately inform clinical decision making with respect to transfusion.