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Mass spectrometry imaging (MSI) is a powerful analytical technique that enables detection, discovery, and identification of multiple classes of biomolecules, while simultaneously mapping their spatial distributions within a sample (e.g., a section of biological tissue). The limitation in molecular coverage afforded by any single MSI platform has led to the development of multimodal approaches that incorporate two or more techniques to obtain greater chemical information. Matrix-assisted laser desorption ionization (MALDI) is a preeminent ionization technique for MSI applications because the wide range of available matrices allows some degree of enhancement with respect to the detection of particular molecular classes. Nonetheless, MALDI has a limited ability to detect and image several classes of molecules, e.g., neutral lipids, in complex samples. Laser desorption ionization from silicon nanopost arrays (NAPA-LDI or NAPA) has been shown to offer complementary coverage with respect to MALDI by providing improved detection of neutral lipids and some small metabolites. Here, we present a multimodal imaging method in which a single tissue section is consecutively imaged at low and high laser fluences, generating spectra that are characteristic of MALDI and NAPA ionization, respectively. The method is demonstrated to map the distributions of species amenable to detection by MALDI (e.g., phospholipids and intermediate-mass metabolites) and NAPA (e.g., neutral lipids such as triglycerides and hexosylceramides, and small metabolites) in mouse brain and lung tissue sections.

Laser ablation electrospray ionization (LAESI) driven by mid-infrared laser pulses allows the direct analysis of biological tissues with minimal sample preparation. Dedicated remote ablation chambers have been developed to eliminate the need for close proximity between the sample and the mass spectrometer inlet. This also allows for the analysis of large or irregularly shaped objects, and incorporation of additional optics for microscopic imaging. Here we report on the characterization of a newly designed conical inner volume ablation chamber working in transmission geometry, where a reduced zone of stagnation was achieved by tapering the sample platform and the chamber outlet. As a result, the transmission efficiency of both large (>7.5 μm) and smaller particulates (<6.5 μm) has increased significantly. Improved analytical figures of merit, including 300 fmol limit of detection, and three orders of magnitude in dynamic range, were established. Particle residence time, measured by the FWHM of the analyte signal, was reduced from 2.0 s to 0.5 s enabling higher ablation rates and shorter analysis time. A total of six glucosinolates (sinigrin, gluconapin, progoitrin, glucoiberin, glucoraphanin, and glucohirsutin) were detected in plant samples with ion abundances higher by a factor of 2 to 8 for the redesigned ablation chamber.

Laser ablation electrospray ionization-mass spectrometry (LAESI-MS) allows for direct analysis of biological tissues at atmospheric pressure with minimal to no sample preparation. In LAESI, a mid-IR laser beam (λ = 2.94 μm) is focused onto the sample to produce an ablation plume that is intercepted and ionized by an electrospray at the inlet of the mass spectrometer. In the remote LAESI platform, the ablation process is removed from the mass spectrometer inlet and takes place in an ablation chamber, allowing for incorporation of additional optics for microscopic imaging and targeting of specific features of the sample for laser ablation sampling. The ablated material is transported by a carrier gas through a length of tubing, delivering it to the MS inlet where it is intercepted and ionized by an electrospray. Previous proof-of-principle studies used a prolate spheroid ablation chamber with the carrier gas flow perpendicular to the ablation plume. This design resulted in significant losses of MS signal in comparison to conventional LAESI. Here we present a newly designed conical inner volume ablation chamber that radially confines the ablation plume produced in transmission geometry. The carrier gas flow and the expanding ablation plume are aligned in a coaxial configuration to improve the transfer of ablated particles. This new design not only recovered the losses observed with the prolate spheroid chamber design, but was found to provide an ∼12-15% increase in the number of metabolite peaks detected from plant leaves and tissue sections relative to conventional LAESI.