Mass spectrometry imaging (MSI) performed under ambient conditions is a convenient and information-rich method that allows for the comprehensive mapping of chemical species throughout biological tissues with typical spatial resolution in the 40-200 μm range. Ambient MSI methods such as desorption electrospray ionization (DESI) eliminate necessary sample preparation but suffer from lower spatial resolution than laser-based and vacuum techniques. In order to take advantage of the benefits of ambient imaging and to compensate for the somewhat limited spatial resolution, a secondary orthogonal separation nested in the imaging scheme was implemented for more selective discernment of tissue features in the spectral domain. Differential mobility spectrometry (DMS), an ion mobility-based separation that selectively transmits ions based on their high-to-low electric field mobility differences, can significantly reduce background chemical interferences, allowing for increased peak capacity. In this work, DESI DM-MSI experiments on biological tissue samples such as sea algae and mouse brain tissue sections were conducted using fixed DMS compensation voltages that selectively transferred one or a class of targeted compounds. By reducing chemical noise, the signal-to-noise ratio was improved 10-fold and the image contrast was doubled, effectively increasing image quality.