Spatially Controlled Molecular Analysis of Biological Samples Using Nanodroplet Arrays and Direct Droplet Aspiration

Mass spectrometry (MS) has emerged as a valuable technology for molecular and spatial evaluation of biological samples. Ambient ionization MS techniques, in particular, allow direct analysis of tissue samples with minimal pretreatment. Here, we describe the design and optimization of an alternative ambient liquid extraction MS approach for metabolite and lipid profiling and imaging from biological samples. The system combines a piezoelectric picoliter dispenser to form solvent nanodroplets onto the sample surface with controlled and tunable spatial resolution and a conductive capillary to directly aspirate/ionize the nanodroplets for efficient analyte transmission and detection. Using this approach, we performed spatial profiling of mouse brain tissue sections with different droplet sizes (390, 420, and 500 μm). MS analysis of normal and cancerous human brain and ovarian tissues yielded rich metabolic profiles that were characteristic of disease state and enabled visualization of tissue regions with different histologic composition. This method was also used to analyze the lipid profiles of human ovarian cell lines. Overall, our results demonstrate the capabilities of this system for spatially controlled MS analysis of biological samples.