Single-filament imaging mass spectrometry lipidomics in Arthrospira platensis.

RATIONALE: Elucidating intra-organismal biochemical and lipid organization in photosynthetic biological cell factories of filamentous cyanobacteria, such as Arthrospira platensis (Spirulina), is important for tracking physiological response mechanisms during growth. Little is known about the filaments' biochemical organization and cellular structure and no label-free imaging techniques exist that provide molecular mapping. METHODS: We applied ultrahigh-resolution mass spectrometry (MS) with matrix-assisted laser desorption ionization (MALDI) imaging to immobilized Spirulina filaments to investigate the localization of lipids across distinct physiological regions. We optimized matrix selection and deposition methods with the goal of facilitating high spatial, and intra-filament, resolution using untargeted multivariate statistical spectral deconvolution across MS pixels. RESULTS: Our results demonstrate an improved two-step matrix application with an optimized procedure for intra-organismal lipid profiling to improve analyte sensitivity and achieve higher spatial resolution. We evaluate several conventional matrices, namely 2,5-dihydroxybenzoic acid (DHB), superDHB (sDHB), 1,5-diaminonaphthalene (DAN), and a 50:50 mix of DHB and sDHB, and compare delineation and pixel-based elucidation of intra-filament lipidomics. We identified a total of 1626 features that could be putatively assigned a lipid-like formula based on database query and 46 unique features, with associated lipid assignments that were significantly distinct in their intra-filament location. CONCLUSIONS: MALDI imaging MS with untargeted statistical spectral deconvolution was used to visualize intra-filament lipidomics organization in Spirulina filaments. Improvements in matrix deposition, including sequential sublimation and pneumatic spraying, increased signal abundance at high spatial resolution and allowed for identification of distinct lipid composition regions. This work outlines a methodology that may be used for micro-ecological untargeted molecular phenotyping.

Interface for direct and continuous sample-matrix deposition onto a MALDI probe for polymer analysis by thermal field flow fractionation and off-line MALDI-MS.

A simple interface based on an oscillating capillary nebulizer (OCN) is described for direct deposition of eluate from a thermal field-flow fractionation (ThFFF) system onto a matrix-assisted laser desorption/ionization (MALDI) probe. In this study, the polymer-containing eluent from the ThFFF system was mixed on-line with MALDI matrix solution and deposited directly onto a moving MALDI probe. The result was a continuous sample track representative of the fractionation process. Subsequent off-line MALDI-mass spectrometry analysis was performed in automated and manual modes. Polystyrene samples of broad polydispersity were used to characterize the overall system performance. The OCN interface is easy to build and operate without the use of heaters or high voltages and is compatible with any MALDI probe format.

Practical implications of ionic strength effects on particle retention in thermal field-flow fractionation.

Modification of ionic strength of an aqueous or non-aqueous carrier solution can have profound effects on the particle retention behavior in thermal field-flow fractionation (ThFFF). These effects can be considered as either advantageous or not depending on the performance criteria under consideration. Aside from the general increase in retention time of particulate material (latexes and silica particles), our experiments indicate improvement in resolution with increases in electrolyte concentration. Absence of an electrolyte in the carrier solution causes deviations from the theoretically expected linear behavior between the retention parameter lambda (a measure of the extent of interaction between the applied field and the particle) and the reciprocal temperature drop across the channel walls. A negative interaction parameter delta(w), of about -0.170 was determined for 0.105- and 0.220-microm polystyrene (PS) latex particles suspended in either a 0.25 or a 1.0 mM TBAP-containing acetonitrile carrier and for 0.220 microm PS in 0.50 and 1.0 mM NaCl-containing aqueous medium. This work also demonstrates that optimum electrolyte concentrations can be chosen to achieve reasonable experimental run-times, good resolution separations, and shifts in the steric inversion points at lower field strengths, and that too high electrolyte concentrations can have deleterious effects such as band broadening and sample loss through adsorption to the channel accumulation surface. The advantages of using ionic strength rather than field strength to effect desired changes are lowered power consumption and possible application of ThFFF to high temperature-labile samples.

Retention behavior of metal particle dispersions in aqueous and nonaqueous carriers in thermal field-flow fractionation.

Until quite recently, theories on thermophoresis of particles predicted very low thermophoretic velocities of metal particles in liquids. This prediction was based on the understanding that the very high thermal conductivities of metals relative to most liquid media resulted in quite low temperature gradients across the metal particle thereby leading to low net force on the particle. In this paper, we report the retention behavior of submicrometer size metal particles of silver (Ag), gold (Au), palladium (Pd) and platinum (Pt) suspended in both aqueous and organic (specifically, acetonitrile and tetrahydrofuran) carrier liquids in thermal field-flow fractionation (ThFFF). The dependence of the metal particle retention on various factors such as particle composition, amount of added electrolyte, carrier liquid composition, field strength, channel thickness, and carrier flow-rate is evaluated and discussed. A comparison in particle retention behavior among equal-sized metal, latex and silica particles is also provided.