The thermal dissipation technique is widely used to estimate transpiration of individual trees and forest stands, but there are conflicting reports regarding its accuracy. We compared the rate of water uptake by stems of six tree species in potometers with sap flow (F S ) estimates derived from thermal dissipation sensors to evaluate the accuracy of the technique. To include the full range of xylem anatomies (i.e., diffuse-porous, ring-porous, and tracheid), we used saplings of sweetgum (Liquidambar styraciflua), eastern cottonwood (Populus deltoides), white oak (Quercus alba), American elm (Ulmus americana), shortleaf pine (Pinus echinata), and loblolly pine (Pinus taeda). In almost all instances, estimated F S deviated substantially from actual F S , with the discrepancy in cumulative F S ranging from 9 to 55%. The thermal dissipation technique generally underestimated F S . There were a number of potential causes of these errors, including species characteristics and probe construction and installation. Species with the same xylem anatomy generally did not show similar relationships between estimated and actual F S , and the largest errors were in species with diffuse-porous (Populus deltoides, 34%) and tracheid (Pinus taeda, 55%) xylem anatomies, rather than ring-porous species Quercus alba (9%) and Ulmus americana (15%) as we had predicted. New species-specific a and b parameter values only modestly improved the accuracy of F S estimates. However, the relationship between the estimated and actual F S was linear in all cases and a simple calibration based on the slope of this relationship reduced the error to 1-4% in five of the species, and to 8% in Liquidambar styraciflua. Our calibration approach compensated simultaneously for variation in species characteristics and sensor construction and use. We conclude that speciesspecific calibrations can substantially increase the accuracy of the thermal dissipation technique.