[3dem] cleaning grids before freezing
Bill & Sue Tivol
wtivol at sbcglobal.net
Fri Aug 22 16:14:51 PDT 2014
Hi Bob,
I use the gold as fiducial markers. If you're doing single-particle,
they provide a high-contrast object for an auto-focus routine to work
with, and if you're doing tomography they are good for image
alignment. I also often add gold to the specimen (for tomography),
but in both cases, I choose a total amount to provide a pretty good
chance of having sufficient beads in the field, but not so many that
they either interfere with images of the specimen or confuse the focus
or alignment routines. I wouldn't think it's effective to prevent
sticking, since that would require a lot of gold.
Yours,
Bill
On Aug 18, 2014, at 8:28 AM, Bob wrote:
> Hi Bill,
>
> Do you use the gold to prevent particles from sticking to the carbon?
>
>
> Bob Ashley
> Electron Microscopist
> Hafenstein Lab
> Penn State College of Medicine
> 500 University Drive
> Mail Code H093 Room C1726
> Hershey, PA 17033
> 717-531-0003 ext. 289256
>
>
>
>
> On Aug 13, 2014, at 1:05 PM, Bill & Sue Tivol <wtivol at sbcglobal.net>
> wrote:
>
>>
>> On Aug 5, 2014, at 9:27 AM, David Gene Morgan wrote:
>>
>>> Hi,
>>>
>>> What are people doing to treat/clean Quantifoil and C-Flat grids
>>> before freezing?
>>>
>>> --
>>> David Gene Morgan
>>> Electron Microscopy Center
>>> 047D Simon Hall
>>> IU Bloomington
>>> 812 856 1457 (office)
>>> 812 856 3221 (3200)
>>> http://bio.indiana.edu/~cryo
>>> _______________________________________________
>>> 3dem mailing list
>>> 3dem at ncmir.ucsd.edu
>>> https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem
>>
>>
>> Dear David,
>> I always glow-discharge for ~1 min and put ~3 uL colloidal gold on
>> the grid, then use just after it dries. If you do not want the
>> gold on your grid, omit that step. When I put the specimen on the
>> grid, I check to see if there is a meniscus on both sides. If so,
>> then the specimen has wet the grid and I am confidant that the
>> plunging will give me a good distribution with specimen in all the
>> holes.
>> Yours,
>> Bill
>>
>>
>>
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>
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