Metalloprotein Microscopy for Quantitative Imaging of Metals and Metallomics Studies in Neurosciences

摘要

Metallomics is a new emerging field addressing the role, uptake, transport and storage of trace metals essential for protein functions. 30percent of the human proteome is metalloproteins. In order to understand the pathophysiology of metalloproteins, metal metabolism and metal-containing deposits and, finally, to facilitate therapeutic interventions, knowledge of the metal distribution in diseased brain compared to controls is fundamental. We propose the creation, development and establishment of the novel metalloprotein microscopy (MPM) by use of a laser microdissection apparatus (LMD) coupled on-line to ICPMS (inductively coupled plasma MS, Thermo Fisher Scientific, Bremen) for direct 2D and 3D bioimaging of metals in the tissue at a scale of 15 and 0.3 micrometers and metallomics studies (German Patent, granted Dec. 2010). LMD as a microscope-based analytical tool using a high precision laser beam to isolate selectively specific region of tissue and cell types from native tissue sections on glass substrate was combine with an ICP mass spectrometer for metal imaging. In MPM a flat laser ablation chamber made of glass was mounted on the microscopic glass slide bearing the sample on the table of the SmartCut Plus LMD (MMI, Zurich). The biological tissue on a regular glass slide inserted in the LMD was ablated using the focused solid-state Nd:YAG laser at a wavelength of 355 nm, laser pulses of 500 ps and with a repetition rate of 500 Hz.