Supplemental data to accompany: **Ultra fast tissue staining using chemical tags** [[http://www.pnas.org/content/111/36/E3805.full|Kohl et al. PNAS (2014)]] and **Second-Generation Drosophila Chemical Tags: Sensitivity, Versatility, and Speed** [[http://www.genetics.org/content/205/4/1399.long|Sutcliffe et al. Genetics (2017)]] ====== Data ====== ====== Image Analysis ====== * [[fly image analysis | Quantification of substrate and antibody penetration into fly brain]] * [[fiber analysis in mouse brain slices | Quantification of substrate and antibody penetration into mouse brain]] ====== Methods ====== Detailed methods are included in the manuscript methods and SI sections. Further information on these topics is available here * [[tagging2014fly-cloning|Detailed cloning steps for fly stocks]] * [[tagging2014flystain |Fly brain staining protocol]] * [[tagging2014mousestain |Mouse slice staining protocol]] * [[tagging2014substrates|Notes on substrates]] * [[tagging2014recflies|Recommended fly stocks]] * [[warping_manual:start|Image Registration]] * [[https://github.com/jefferis/AnalysisSuite|Morphological neuroanatomy in R]] aka AnalysisSuite =====Information, Discussions & Enquiries===== Please send any technical enquiries to tissuetagging@googlegroups.com to engage all authors of the paper. We will also use this list to update you about any improved stocks that we make. You can freely send email to that address but you must sign up here https://groups.google.com/forum/#!forum/tissuetagging to read existing discussions. =====Resources===== * There are some notes on the fly lines and reagents {{:si:tagging:tissuetaggingflies.pdf|here}} that we recommend you read before contacting Greg to request any flies. * Fly Stocks are freely available from the [[http://flystocks.bio.indiana.edu/Browse/misc-browse/chemtag.php|Bloomington stock centre]] * Jefferis lab members can order these fly stocks ("PT stocks") from Glynnis Johnson at Genetics (gh19@hermes.cam.ac.uk) * Virus constructs have been deposited with Addgene (#60098 and 60099) and should be available by Dec 2014 at the latest:

===== Links ===== [[http://biorxiv.org/content/early/2014/05/19/005298|public preprint]] from http://biorxiv.org/ (includes a complete protocol for fly brain staining) [[https://www.neb.com/tools-and-resources/feature-articles/snap-tag-technologies-novel-tools-to-study-protein-function|NEB SNAP/CLIP-tag]] [[http://www.promega.co.uk/products/pm/halotag-technology/halotag-technology/|Promega Halo-tag]] [[http://www.activemotif.com/catalog/122/ligandlink-universal-protein-labeling|Active Motif TMP-tag]] (we used the covalent version)