Paulina Dominika Rakowska oral presentation (PB3-Thu1-2-5)
Cryo-SIMS – metrology of biological sample preparation methods for preservation of cell ultrastructure and chemistry.
National Physical Laboratory, Hampton Road, TW11 0LW Teddington, United Kingdom
With the potential of high-throughput, high-resolution and high-sensitivity label-free imaging in 3D, secondary ion mass spectrometry imaging methods are, arguably, ones of the most powerful techniques for high-resolution chemical imaging of biological samples. However, there are some critical limitations for these analyses. As the high-performance SIMS instruments require high vacuum, a careful consideration of sample preparations is often needed. For example, advanced methods are necessary to prepare and measure complex hydrated bacterial biofilm structures. Also, in the pharmacological imaging of potential drug candidates at their targets, the positioning of water soluble drug compounds within cells or tissues can be altered by pre-treatment processes such as drying, resin-embedding or histological fixation. Advanced cryo-preparation methods are necessary for immobilisation of water in these samples to prevent the ultrastructural reorganisation and the loss or translocation of water-soluble molecules, to circumvent the use of chemical fixation and to enable their analysis in high-vacuum of mass spectrometry instruments.
The UK’s National Centre of Excellence in Mass Spectrometry Imaging (NiCE MSI) at NPL has a special focus on the development of advanced solutions to challenging measurements. Our recently innovated 3D OrbiSIMS instrument has the capability to handle and measure cryogenically-prepared samples. The instrument is equipped with a vacuum cryo transfer system that is compatible with cryo-SEM and cryo-TEM. A shuttle chamber allows the interchange of samples, in vacuum and cryogenically, between cryo-preparative equipment and the 3D OrbiSIMS instrument.
This presentation will show our recent developments of the cryo-SIMS methodologies. Different sample cryo-preparation techniques will be compared, such as the analysis performed on frozen-hydrated vs. frozen-dehydrated mammalian cells. The application of cryo-SIMS to a range of biological samples including cells, bacteria, biofilms and organic reference samples will be presented. Focus will be given to the use of different types of cryo-protectants, often required for the vitrification of thicker samples such as biofilms, by high-pressure freezing and their effects on SIMS analysis.