[3dem] cleaning grids before freezing

[Bill & Sue Tivol]

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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