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Specimen Processing: allergy to histological solvents Aſt er decades of using standard organic solvents for paraffi n- section histology, I fi nd that I’ve become highly allergic to the fumes. T ese include commercial preps like Citrosolv and Hemo-De, as well as xylene and toluene, which cause dizziness and pounding headaches. T is has made staining and cover slipping very diffi cult, even with hood. Can anyone recommend alternative solutions for (a) dewaxing and (b) cover slipping with Permount? Daniel Blackburn dblackburn2000@yahoo. com Sat Feb 20

I assume you’re doing these steps in a fume hood. One thing we’ve found helpful for people who are sensitive to solvent fumes are charcoal-lined dust masks. Search for “charcoal fi lter masks”. We use the 3M type (but not for the prices here - we got ours through Fisher Scientifi c). So far, they’ve worked fi ne. Looks like you’ve already tried the various xylene replacements. Phil Oshel Sun Feb 21

A serious problem. By listing, I have come with suggestions. I have long used a Gum Damar formulation that I cannot offer, because I have prepared it in xylene. Note: for those who might be interested—or users of Damar—I have a liter that I will never use more than a few ml’s over my reasonably extended age - now 76. Further, I don’t do paraffin any more. The suggestions: If you can arrive at a solvent for Damar that is miscible with xylene/ toluene, then it would be possible that you could prepare your own Damar. Note: Damar applied in the 1960s still has no sign of drying/cracking/crystalizing. Source:

com/shop/950-damar-resin-crystals-graded/ . You might test a hydrophilic mountant. Source:

microscopy/products/histology/mounting_media.aspx . If your allergy is broad-band, then you may have to resort to the latter. If it is more specific - just for those you mentioned - then your options might include Canada Balsam that is very natural, but also often found with benzene or xylene or toluene. I have some that will never be used as it requires some solvent that might not raise a sneeze. Frederick C. Monson Sun Feb 21

Image Processing:

batch convert emi to other format We have an FEI TEM microscope. T e STEM images Acquired in

TIA soſt ware are saved in emi format. I can open them in TIA soſt ware, and can only export them one by one. T at is quite annoying. Is there anybody who has any idea how to batch convert the images? Hongbing Yu T u Feb 18

TIA folder export: - Open menu Tools\Components. - Click “Install”. - Type “folderexport.wsc”. - “ok”. Now that this component is installed, there will be an additional menu in the processing tasks. Check settings and hit Export. Wim Hagen T u Feb 18

Microtomy: cryo-ultramicrotomy of rubber sections

Can anyone recommend a technique for collecting rubber sections during cryo-ultramicrotomy that enables the sections to lie fl at (i.e. as wrinkle free as possible) on a TEM support grid? I currently collect my sections off the back of my cryoknife using a sucrose droplet, but as the sections warm coming out of the cryochamber on the droplet they contract and wrinkle before being deposited on a TEM grid (with or without carbon coating). Chamber temperature is -120°C. Any ideas are appreciated. Anand Badami Mon Feb 8

2016 May •

If the wrinkling happens during warming of the droplet, could it be because of the section surface characteristics? Adding salt to the sucrose should help if the surface is charged; multivalent ion salts would be more suited than monovalent. However if the surface is not charged but hydrophobic adding salt would probably increase the wrinkling, and in that case adding a hydrophobic component might help, such as serum albumin. Some detergents can do both. Jan Leunissen Mon Feb 8

LM: iPhone microscope

Someone has asked me about microscope attachments for iPhone. I’ve seen lots of ads, but I haven’t looked closely at any of the products. Do any of you have opinions about products that turn your iPhone or Android into decent microscopes? I’m not sure if she wants to go with a stand and slides, or just take nicely magnifi ed images on the go. I think the former. Tina (Weatherby) Carvalho tina@pbrc.hawaii. edu Wed Jan 13

Here at PNNL we’ve actually developed a low-cost (~$1) cellphone camera that magnifi es up to 1000×, depending on the specifi cation. Our lab has freely released the blueprints for 3D printing or you can order pre-manufactured lenses that clip onto pretty much any smartphone. We use them frequently in our demos to high school and middle school children and they work quite well once you get the hang of them. Here’s a link to more details: http://availabletechnologies. . Steven R. Spurgeon steven.spurgeon@ Wed Jan 13 T ough not as cheap as the PNNL design, some colleagues of mine have used this inexpensive, hardware-store-materials-based home-built design for outreach and liked it: 10-Smartphone-to-digital-microscope-conversion/ Tyler Harvey trh@ Wed Jan 13

I got 17 replies to my query about turning an iPhone into a microscope, and no two were the same. Most of them were along the lines of “I saw this one on the Internet...”. If anyone is particu- larly interested, I can send a list of links or forward some of the responses. Here is the one that the person who requested feedback is looking at (and is not one of the ones any of you mentioned): . It holds a slide and I think has a light source. Tina (Weatherby) Carvalho Fri Jan 15

TEM: bias and gain correction on camera

We have a SIS Megaview III camera (SIS, then Olympus and now I see it is EMSIS “again”) for over 10 years, a real workhorse, always performing well without complaining. Because it works so well I tend to forget about it and now I am just wondering if I need to update the gain and bias corrections (the correction pictures)? I can’t remember the last time I did it but it is a long time ago. I know that it is not a lot of work to do it, I just wondered if there is any reason to do this regularly and if yes, what are the time intervals. Stephane Nizets T u Feb 18

Since the blacklevel (“noise”) and gain images are a necessary step for getting an artefact-free and evenly background, I think you should do this short procedure (depends what version of Analysis or ITEM soſt ware you are using) oſt en. When I am installing or updating cameras at the customer´s site I ask for the most used magnifi cation / spot size / apertures setting they use and do the correction images with this setting. But: I also know a customer who is doing these image sets prior to each high-res image he uses for publication (makes sense). Stefan Diller T u Feb 18


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