Posted by
George McNamara on
URL: http://confocal-microscopy-list.275.s1.nabble.com/disinfecting-a-microscope-tp7584530p7584537.html
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopyPost images on
http://www.imgur.com and include the link in your posting.
*****
Hi Anil,
Eyepieces ... get the camera image in focus, then adjust the eyepieces
to match YOUR eyes. the IX71 should have adjustable eyepieces. If you
wear glasses (or contacts) adjust either on (or off) and look that way
consistently. If you can't get the eyepieces to match the camera, either
the camera was installed incorrectly or you need to new glasses.
Staining ... don't overstain, wash enough to get rid of background, use
"best" mounting medium -- probably not PBS. that said, a lot simpler if
you start with PBS.
Don't spend the rest of your life making coverglass-slide preparations.
It's an inverted microscope: use imaging dishes. I mostly use
http://glass-bottom-dishes.com/gbcustomerpriceweb.pdfP35G-1.5-20-C
(20 mm diameter glass imaging area, #1.5 coverglass).
If comparing live cell cultures, I often use the Greiner Bio-One
CellView quadrant dishes ("4 compartments"),
https://shop.gbo.com/en/row/articles/catalogue/articles/0110_0110_0040_0020/(prolonged live cultures, i usually overlay Sigma-Aldrich M8410 mineral
oil, since my PECON litte incubator thing allows the dishes to dry out)
Nikon also sells "Hi-Q" quadrant dishes, with a funky upper plastic
thingy to (try to) reduce the meniscus effect on transmitted light
imaging (one of many reasons I pay more attention to the
epi-fluorescence than the transmitted light channel).
For starting, I suggest 20x/0.75 (20x/0.70 NA if Olympus did not sell
you one with that extra 0.05), and change fields of view pretty often to
avoid photobleaching.
I might - or not - have useful tips in the MPMicro download available at
http://works.bepress.com/gmcnamara/2/(one tip i need to mention here; please DO NOT HIT THE PRINT BUTTON -
thanks).
I haven't updated MPMicro in a while, so this part may be out of date
(that is, more than 50% of URLs dead links), my favorite section is
"Accessorize". The section I still use the most is the one with the
table of refractive indices.
For any listservites in (or near) Houston: Marty Chalfie is speaking at
MDACC on Jan 7 and 8 (2016). Should be fun.
George
p.s. if colleagues want a look, simpler to use the monitor and focus
with live camera, than taking turns readjusting the eyepieces all the time.
For "wet mounts" with fixed cells i tend to use 80% glycerol:20% PBS, pH
7.4. Work ok for most fluorophores.
On 12/16/2015 1:07 PM, Anil Poudel wrote:
> *****
> To join, leave or search the confocal microscopy listserv, go to:
>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> Post images on
http://www.imgur.com and include the link in your posting.
> *****
>
> Hello there,
>
> I am new to fluorescence microscopy and trying some F-actin staining kit (Phalloidin 555 from Invitrogen) but having hard time to get very clean filamentous picture and also unable to get the image on screen as it appears through eyepiece. I am using Olympus IX71 with qclick monochorome camera and cellsens software. Any suggestion would for optimization is highly appreciated. Thanks in advance.
>
> Anil
> ________________________________________
> From: Confocal Microscopy List <
[hidden email]> on behalf of Cromey, Douglas W - (dcromey) <
[hidden email]>
> Sent: Wednesday, December 16, 2015 2:01 PM
> To:
[hidden email]
> Subject: disinfecting a microscope
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> Post images on
http://www.imgur.com and include the link in your posting.
> *****
>
> What is an appropriate method for disinfecting a microscope used for live cell imaging (or in a BSL2 facility)? 10% bleach is too strong, so what do people use? Thinking of core facility users (who can be poor at following directions), what is a relatively fool-proof way to decontaminate and not damage motorized stages, and other delicate parts?
>
> If you have posted protocols, can you send the links? Do you have live cell imaging policies for your facility that you would be willing to share?
>
> Doug
>
> ------------------------------------------------------------------------------------------
> Douglas W. Cromey, M.S. - Associate Scientific Investigator
> Dept. of Cellular & Molecular Medicine, University of Arizona
> 1501 N. Campbell Ave, Tucson, AZ 85724-5044 USA
>
> office: UACC 0914A email:
[hidden email]<mailto:
[hidden email]>
> voice: 520-626-2824 fax: 520-626-2097
>
>
http://swehsc.pharmacy.arizona.edu/micro> Home of: "Microscopy and Imaging Resources on the WWW"
>
> UA Microscopy Alliance -
http://microscopy.arizona.edu<
http://microscopy.arizona.edu/>
>
--
George McNamara, Ph.D.
Single Cells Analyst
L.J.N. Cooper Lab
University of Texas M.D. Anderson Cancer Center
Houston, TX 77054
Tattletales
http://works.bepress.com/gmcnamara/42http://works.bepress.com/gmcnamara/75https://www.linkedin.com/in/georgemcnamara