Tissue preparation protocols imaging

Two variations of conventional SIMS have evolved. In one, known as matrix-enhanced SIMS, the MALDI sample preparation protocol is used to coat the sample surface with an organic acid (e.g., 2,5-dihydroxybenzoic acid) or other suitable matrix material to improve ionization efficiency [24,25], This approach has been used to image brain tissue samples from freshwater snails (Lymnaea stagnalis). Another variation employs a coating of a thin layer of gold or other metals to enhance analytical signals [26], This method, known as metal-assisted SIMS, provides images with improved spatial and chemical resolution. 

Croft, William J. Under the Microscope A Brief History of Microscopy. Hackensack, N.J. World Scientific, 2006. Traces the microscope from early beginnings to modern instruments, discussing how each works. Hewitson, Tim D., and Ian A. Darby, eds. Histology Protocols. New York Humana Press, 2010. This laboratory manual looks at tissue preparation and staining, with explanations of complex procedures. Ovalle, William K., and Patrick C. Nahirney. Netter s Essential Histology. Philadelphia Saunders/Else-vier, 2008. This atlas covers cells and tissues and the major bodily systems. Features a great collection of images by Frank H. Netter. 

Quantitative images of elements can be obtained using prepared matrix-matched laboratory standards for calibration as demonstrated in several previous papers (I, 21, 42). The preparation protocol for synthetic matrix-matched laboratory standards for the calibration of imaging LA-ICP-MS for biological tissues is summarized in Fig. 3.3. In general, different synthetic laboratory standards with elements of interest in well-defined concentrations were prepared. For example, five slices of the same biological tissue were spiked with selected standard solutions at... 

Successful application of this experimental approach depends on several factors synthesis of high-quality hybridization probes, appropriate fixation of the sample, the hybridization procedure, and the fluorescence microscopy approach used to image the specimen. In adapting the technique of three-dimensional in situ hybridization to different organisms and tissue types, the simplest and most invariant aspect of the technology has proved to be the hybridization procedure. Probes must be developed on a custom basis to address the particular questions of the investigator, and equally crucially, fixation conditions need to be adapted with special attention to the physical attributes of the individual specimen. However, once appropriate preparation conditions are established for a particular type of sample, it has been unnecessary to reoptimize the basic hybridization protocol. We discuss each of these experimental issues separately below. 

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