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Edited by Thomas E. Phillips University of Missouri


phillipst@missouri.edu


Selected postings from the Microscopy Listserver from November 1, 2017 to January 1, 2018. Complete listings and subscription information can be obtained at http://www.microscopy.com . Postings may have been edited to conserve space or for clarity.


Microscopy Listserver Statistics:


Welcome to another year of operation of the Microscopy ListServer—a free service to the worldwide microscopy community, sponsored jointly by your Friendly Neighborhood SysOp and the Microscopy Society of America. During 2017, the ListServer delivered your messages to more than 4,350 subscribers around the world, with minimal hassles (that I know about). For those of you who are statistics junkies, this year you generated ~250+ Gbits of email traffi c, and ~2.55 million email messages were sent out by my tired and old little server. Up a bit from last year, but still a steady fl ow.


As usual you don’t want to know how much junk mail and spam


has been fi ltered out—far more than you might expect. Apologies to those who have problems with my fi lters. T e complete Microscopy ListServer Archives for (approxi- mately) 2017–1994 are online at http://www.microscopy.com . A few important reminders: • If you leave on vacation/holiday use the online form to UNSUBSCRIBE your email address from the Listserver. T e out-of-offi ce/on-vacation autoreply messages are a real nuisance to posters.


• Do not reply to messages with the return address of MicroscopyListserver-noreply-at-microscopy.com. T ese are messages forwarded usually from the WWW posting form. T ey do not go back to the poster but rather into a black hole, which I rarely check. If you see a message that has this “no-reply” return address, please post your reply/comment/answer to: microscopy@ microscopy.com , or if you wish to reply privately, look at the username in the body of the message; the originator’s email address is usually listed therein.


• As always if you have questions about suitability of postings or are having problems, feel free to contact me at zaluzec@microscopy.com . Nestor Zaluzec zaluzec@microscopy.com Sun Jan 1


Specimen Preparation: coating for Carbon evaporator bell jar


In a previous post, I asked what everyone was doing for coating the bell jar on your carbon evaporator, now that Bell Bright isn’t going to be available. It seemed the consensus was to use dish soap as a replacement. Well, I’ve exhausted my hoard, and like a dummy, I didn’t ask for how you did that. So the obvious questions, are what are you using instead of Bell Bright and how do you apply it? Matthew Duley duleyml@ muohio.edu Fri Dec 1


It is very simple and inexpensive; clean your bell jar, electrodes, and insulators as best you can then wipe the inside with a piece of


60


paper towel that has had 5-10ml of dishwashing detergent applied to it. Wipe the whole of the glass, metal and insulator surfaces so that there is a very thin smear of detergent applied. Pump down the bell jar to dry the system out. T e system is ready to use! Next time you need to clean the system all you need do is wash all surfaces with warm water, dry them off with a paper towel, then apply the detergent again. John Nailon jvnailon@gmail.com Sun Dec 3


I have a coater I’m refurbishing and wondering what the purpose is for this detergent coating? Sylvain Poudrette spoudrette@videotron. ca Mon Dec 4


Every time you use the evaporator to coat a sample with carbon or metal, everything gets coated. I am alluding to the inside of the glass bell jar, all of the electrodes and all of the insulated connections. If you do not clean the system periodically, then you will start to have slow pump down times, inability to see inside the bell jar and potential short circuits across the insulated connections within the bell jar. John Nailon jvnailon@gmail.com Sun Dec 10


I have the Denton floor model carbon evaporator, and I use a very small dab of metal polish on a paper towel, within a small area of the jar. I immediately chase with 100% ethanol, then move to another small region of the jar. I have never had vacuum issues cleaning the bell jar like this. Michael Delannoy mdelann1@jhmi. edu Mon Dec 4 We off er BellShine at https://www.laddresearch.com/vacuum- equipment/bell-shine that should work for you. Call and ask for Mike Bouchard if you want to discuss it. Disclaimer: Ladd research sells this product and other products for use in Microscopy Labs. JD Arnott jd@laddresearch.com Mon Dec 4


Specimen Preparation: chick brain for Vibratome sectioning


I am doing brain electroporations with an RCASB plasmid, and


I wanted to visualize how effi cient my electroporations were. Currently, I am embedding my heads in 3% agarose at 37 ºC between 15-20 minutes, cutting the block fairly close to the brain, and a fi nal embedding in 6% agarose at 4ºC for at least 20 minutes. T e smallest sections I have been able to slice is 300 microns; however, I would like to get down to 200 microns. When I have attempted to go below 300 microns, the tissue is destroyed, which I can see that it’s lacking structural support from how I am embedding. My question: What would be a more effi cient way to embed my samples so I can section between 100-200 microns? Garrett Driscoll driscollg2@winthrop.edu Tue Dec 5


First, are you removing the meninges before embedding the


brains? T ey can interfere with the agarose. Second, this may/may not help — I embedded mormyrid brains, not vertebrate. Try 18% (wt:vol) 100-175 bloom gelatin instead of agarose. Liquefy the gelatin and place tissue in the gelatin in a 55-60ºC oven for 1.5-2 hours. Remove from oven, blot, place in fresh gelatin & put in the oven for 15-30 minutes, remove from oven, blot, embed in fresh gelatin but do not put back


doi: 10.1017/S1551929517001377 www.microscopy-today.com • 2018 March


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