This work investigates the applicability of fast detectors to the technique of microscope-mode imaging mass spectrometry. By ionising analyte from a large area of the sample, and projecting the desorbed ions by the use of ion optics through a time-of-flight mass spectrometer onto a two- dimensional detector, time- (and hence mass-) dependent distributions of ions may be imaged. To date, this method of imaging mass spectrometry has been limited by the ability to image only one mass window of interest per experimental cycle, limiting throughput and processing speed. Thus, the alternative microprobe-mode imaging mass spectrometry is currently the dominant method of analysis, with its superior mass resolution. The application of fast detectors to microscope-mode imaging lifts the restriction of the detection of a single mass window per experimental cycle, potentially decreasing acquisition time by a factor of the number of mass peaks of interest. Additional advantages include the reduction of sample damage by laser ablation, and the potential identification of coincident eo-fragments of different masses originating from the same parent molecule. Theoretical calculations and simulations have been performed confirming the suitability of conventional time-of-flight velocity-mapped ion imaging apparatus for imaging mass spectrometry. Only small modifications to the repeller plate and laser beam path, together with the adjustment of the accelerating potential field, were required to convert the apparatus to a wide (7 mm diameter) field-of-view ion microscope. Factors affecting the mass and spatial resolution were investigated with these theoretical calculations, with theoretical calculations predicting a spatial resolution of about 26μm and m/m of 93. Typical experimental data collected from velocity-mapped ion imaging experiments were collected, and characterised in order to provide specifications for a novel time-stamping detector, the Pixel Imaging Mass Spectrometry detector. From these data, the suitability of thresholding and centroiding on the new detector was determined. Initial experiments using desorptionjionisation on silicon and conventional charge-coupled device cameras confirmed the correct spatial-mapping of the apparatus. Matrix-assisted laser desorptionjionisation techniques (MALDI) were used in experiments to determine the spatial and mass resolutions attainable with the apparatus. Experimental spatial resolutions of 14.4 μm and m/m of 60 were found. The better experimental spatial resolution indicates a higher di- rectionality of initial velocities from MALDI desorption than used in the theoretical predictions, while the poorer mass resolution could be attributed to limitations imposed by the use of the phosphor screen. Proof-of-concept experiments using fast-framing cameras and the new time-stamping detectors confirmed the feasibility of multiple mass acquisition in time-of-flight microscope mode ion imaging. Mass-dependent distributions were acquired of different pigment distributions in each experimental cycle. Finally, spatial-mapped images of coronal mouse brain sections were acquired using both conventional and fast detectors. The apparatus was demonstrated to provide accurate spatial distributions with a wide field-of-view, and multiple mass distributions were acquired with each experimental cycle using the new time-stamping detector.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:564395 |
| Date | January 2012 |
| Creators | Yuen, Wei Hao |
| Contributors | Brouard, Mark |
| Publisher | University of Oxford |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
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