Multidimensional analysis of single algal cells by integrating microspectroscopy with mass spectrometry

Abstract

We demonstrate a facile label-free approach for performing multidimensional chemical analysis on individual single-cell organisms by combining optical, fluorescence, and Raman microspectroscopy with matrix-free laser desorption/ionization mass spectrometry (MS). Single unicellular algae are seeded on a bare stainless steel plate and analyzed microspectroscopically. This provides information on the content and distribution of photoactive species, such as β-carotene, as well as chlorophyll and other components of the photosynthetic apparatus. Exactly the same cells are then analyzed by mass spectrometry in the negative ion mode. Phospholipid species are readily ionized by laser desorption/ionization of intact cells, without the need for an auxiliary matrix. This not only facilitates sample preparation but also preserves high spatial resolution and high sensitivity. Using this method, we were able to study the content and arrangement of proplastids and photosystem components, as well as the amounts of various phospholipid species in individual algal cells. The methodology can be used in the fundamental biological studies on these unicellular organisms, which require information on the internal structure as well as the chemical composition of individual cells.