In this short communication, we report simultaneous measurement of CO and OH in flames using a sin- gle femtosecond (fs)-duration laser source. Two-photon excitation of CO A(1) Pi <-<- X-1 Sigma(+)(3,0), (4,0) and (5,0) bands and single-photon excitation of OH A(2)Sigma(+) <- X-2 Pi (1,0) band are achieved simultaneously using one fs-duration laser pulse near 283 nm. Subsequently, crosstalk-free, laser-induced fluorescence (LIF) emissions are detected from the v'=3, 4 and 5 upper vibrational levels of CO in the 200-230 nm region, and (1,1) and (0,0) bands of OH near the 310-nm region. A detailed spectroscopic investigation of CO in a gas cell is followed by simultaneous detection of CO and OH in C2H4/air flames. The measured CO and OH profiles as a function of flame equivalence ratio agree well with equilibrium calculations. The present study extends the applicability of broadband fs-LIF imaging of OH in flames, and reveal an alternate method to obtain heat release rate in turbulent flames via simultaneous OH and CO measurements. (C) 2020 The Combustion Institute. Published by Elsevier Inc. All rights reserved.