Morphologically compatible mass spectrometric analysis of lipids in cytological specimens

Modern lipid analysis requires mass spectrometric techniques, though to date these have been developed and applied primarily to histological serial sections. As such, there has been little emphasis on using mass spectrometry in such a way that the same specimen can yield both chemical and morphological information. Here, we present a mass spectrometric method that enables measurement of lipids from cells on cytospin slides in a way that preserves the cells for subsequent cytomorphologic evaluation.

Standardized cultures of MDA-MB-231, a breast cancer cell line, were prepared as cytospins and subjected to analysis using a Prosolia Flowprobe sampling and ionization source attached to a Thermo Scientific Quadrupole-Orbitrap mass spectrometer. Chemical compositions were deduced with accurate mass measurements and fragmentation of high intensity peaks to further determine chemical structure. After mass spectrometry, the slides were stained and cover-slipped, and the cells were reviewed for cytomorphologic features of breast cancer. These were compared to control slides of the same cellular concentration that had not been subjected to this analysis.

Spectra from samples of all cellular concentrations demonstrated characteristic qualitative features that were discovered to represent phosphatidylcholines, phosphatidylglycerols, and phosphatidylserines with fragmentation and accurate mass matching. Cytomorphologic analysis demonstrated excellent preservation of the cells subjected to the Flowprobe analysis.

Direct extraction, ionization, and identification of lipids is possible from cytologic preparations in such a way that the analyzed material is preserved and useful for subsequent microscopic analysis.