NetNotes
have WDS installed, and I understand it is aimed at 10 mm. So, I will probably just leave things as they are and bring up the current a bit more. Now to the other question of performance and resolu-
tion as a function of size. It is true that all other things being equal, you may give up a little energy resolution by going to a bigger detector. It is harder to find a crystal of the same, good resolution when it must be 8×larger, but it can be done. Tey will probably charge a premium for it. You probably want to compare spectra on your real samples. I choose to back off on the process time to push more counts at the same deadtime. So, I am already sacrificing some resolution. I am glad that I started with a better detector. Tere are times that I want better resolution, so I increase the process time and cut the count rate—or I switch over to the WDS. Warren Straszheim
wesaia@iastate.edu
Dipping Objectives - Water or PBS? Confocal Listserver Does anyone have experience or know if water dipping
objectives (specifically the Zeiss W Plan-Apochromat) are designed for immersion in distilled deionized water or PBS or if they are designed to perform well in both based on refractive indices? Adam Glaser
adamkglaser@gmail.com
I’m working with this lens on in vivo samples. We use both
and I don’t see much difference, but I work with a temperature- controlled chamber so most of the time I use ultrasound gel. Pascal Weber
pascal.weber@
univ-amu.fr
Do you find a difference in refractive index between PBS
and water? I think they are both 1.33. If they have the same refractive index then the objective should perform identically with both. Sylvie le Guyader
sylvie.le.guyader@
ki.se
Te salt doesn’t make that much of a difference. One detail
is to remind users to rinse the lens off with pure water aſter using PBS. I made a “house call” to a lab that was having prob- lems using one of their patch rigs. Aſter checking all their fil- ters and prisms I happened to look down at the objective and found a perfect window of dried salt sitting over the face of the objective. One lengthy freshwater soak later and everything was fine. Craig Brideau
craig.brideau@
gmail.com
A hint following on Craig’s spur from the original PBS
refractive index question. Caked-on salt films on water immer- sion objectives that have formed from repeated exposure to PBS buffers can be removed by dipping them in 0.1 N acetic acid solution for ∼10 min and rinsing with no obvious deleteri- ous effect. Mark Sonders
mss2110@columbia.edu
Tanks everyone. I also see online that PBS should have a
RI of 1.335. If this is true, then I agree it is a very small differ- ence. Measuring some PBS in our lab on a refractometer I got a reading of 1.341, which seemed like it could start to have an impact. I will try to contact Zeiss and see what they designed the objective for, otherwise I’ll just go with DI water for now! Adam Glaser
adamkglaser@gmail.com
74
Camera Bubbles Confocal Listserver Has anyone seen/experienced this issue before? Minutes
into using the camera, strange “boxes” appear in the center of the image and spread to cover the whole field, eventually turn- ing into “bubbles.” Tese “boxes” are the same size no matter the magnification used. Any help/advice is greatly appreciated! Laurel Ballantyne
ballantl@queensu.ca
Do you see these features in the sample aſter observing
with the binocs (your eyes) and not using the camera? If so, I’d suspect your mounting medium. Philip Oshel oshel1pe@
cmich.edu
I have never seen boxes before, but if the artifact appears
to grow under the beam then the section is probably still wet. Otherwise perhaps a salt contamination. Michael Delannoy
mdelann1@jhmi.edu
Test whether you also see it if you only image glass. We
had the same with a Hamamatsu camera and got a replacement due to condensation in the camera. Tanneke denBlaauwen
t.denblaauwen@
uva.nl
Definitely a broken seal in the camera with condensation
on the inner surface of the protective window. If you leave the camera on for a few days, it may vanish. Elmar Tews (email unavailable)
If you rotate the camera while viewing live, do the bub-
bles move with the image? If not, then the bubbles are on the camera and likely result from condensation from a failed seal within the camera. Brian Kain
bkain@hamamatsu.com
We did experience this once with a Zyla camera. No, the
condensation won’t go away and the camera will need a fac- tory refurbishing. We then put a dehumidifier right next to it to prevent it from happening again. John Ho-chun hclaiac@
connect.ust.hk
Tey’re only there through the camera, not when looking
through the eyepieces. Tanks for the input, though quite sad. When I rotate the camera, the squares/bubbles don’t move. Tanks for the info! Tanks so much, everyone, for all the info! We’ll look in to sending the camera back for repair. Laurel Ballantyne
ballantl@queensu.ca
CMOS Inquiry for Camera Gurus Confocal Listserver Dear camera gurus, our Andor Zyla 5.5 sCMOS camera
shows condensation on the chip when we cool it. Andor told us this might be caused by a backfill issue and it costs ∼$1,000 to take a look, possibly a 4–6 week turnaround, and probably more money to fix the problem. I have a couple of questions and I am also looking for a little advice. 1) I remember hearing a little bit about cameras holding vacuum, but not about this backfill prob- lem. Anyway, what is backfill and how does it go wrong? Is it an inert, dry gas around the sensor that may now have some moist
www.microscopy-today.com • 2020 May
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