https://www.osa-opn.org/home/book_reviews/2020/0320/book_review_of_multiphoton_microscopy_(neuromethod/?feed=BookReviews

I recommend this exciting book, replete with full-color images, because of its comprehensive, tested protocols for the study of live neural tissue at the cellular and the subcellular levels. The limitations of the various microscopes and specimen labeling are clearly stated. The use of genome editing with CRISPR/Cas9 to label endogenous proteins offers an important advance to mitigate the problem of overexpression of genetically encoded fluorescent probes. The problems of phototoxicity, photobleaching of probes, and motion artifacts are addressed for each protocol. The contributors detail the research questions asked, the design and operation of the optical instruments, the specimen preparation, the use of new electrochromic ANNINE dyes, acquisition of the images, and the critical evaluation of the data. The control experiments, cautions, calibrations and trade-offs are discussed. The detailed listings of sources, instruments, components, open-access software and MATLAB code, as well as a comprehensive listing of modern references, are commendable.