By Lin He
Mass Spectroscopy Imaging (MSI) has emerged as an allowing strategy to supply perception into the molecular entities inside cells, tissues and whole-body samples and to appreciate inherent complexities inside of organic metabolomes. In Mass Spectrometry Imaging of Small Molecules: tools and Protocols, specialists within the MSI box current innovations for second and 3D visualization and quantification of a wide range of small molecular species found in biologically proper samples. Chapters offer unique operational directions from pattern instruction to approach choice, from comparative quantification to structural identity and from info assortment to visualization of small molecule mapping in advanced samples. Written within the profitable Methods in Molecular Biology sequence structure, chapters comprise introductions to their respective themes, lists of the required fabrics and reagents, step by step, with no trouble reproducible protocols and notes on troubleshooting and fending off recognized pitfalls.
Authoritative and simply available, Mass Spectrometry Imaging of Small Molecules: equipment and Protocols goals to deliver the speedily maturing tools of metabolic imaging to existence technological know-how researchers and to lessen technical intimidation in adapting new technological structures in organic research.
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Extra resources for Mass Spectrometry Imaging of Small Molecules
For imaging applications, ideal section thickness should be the width of one cell; 8–20 μm thickness is appropriate. , less tearing or folding) but thinner sections typically result in better sensitivity. 1. Trim the root nodule from the plant, leaving 3–4 mm of root attached to the nodule (1–2 mm on each side of the nodule). 2. Immediately after dissection, submerge the tissue in a small plastic cup of gelatin (see Notes 2 and 3). 3. Use forceps to orient the tissue as desired. 4. Once the tissue is stuck to the bottom of the cup, use a syringe or pipet to cover the tissue with more gelatin (approximately 3–5 mm higher than the tissue) (see Note 4).
It is important to keep the same orientation of the sample between the microscope observation and the MALDI MSI acquisition in order to easily correlate the images. 6. 4800 MALDI TOF/TOF (AB Sciex) works with a fixed laser focus. , larger crater size. , proteomics for which high sensitivity is required versus MSI for which high lateral resolution is required). References 1. Spengler B, Hubert M, Kaufmann R (1994) In: Proceedings of the 42nd Annual Conference on Mass Spectrometry and Allied Topics, Chicago, IL, p 1041 2.
Key words Imaging mass spectrometry, Mass spectrometric imaging, MALDI, TOF/TOF, Legume, Metabolite, Mass spectrometry 1 Introduction Legumes are extremely important to the agriculture industry because they can grow in a wide variety of agroecological conditions and have developed the unique ability to fix their own nitrogen through their symbiotic relationship with soil bacteria known as rhizobia . Decades of research on legumes have been dedicated to deciphering the metabolic networks involved in nitrogen fixation [2, 3].