[3dem] electron dose & data collection strategy for Titan Krios, 300kV, Falcon back thinned

[m.maletta at nki.nl]

Dear Sjors,
I never heard about kicking out the fist two frames. Do you have any idea why is so?
It is somewhat related to the charge-peeling of the exposure area?
Best
Max
__________________________________
Massimiliano Maletta
post-doc in
NKI Netherlands Cancer Institute -Antoni van Leeuwenhoek Hospital Plesmanlaan 121 B6 1066CX Amsterdam
group of Peter Peters
www.nki.nl/research/peters/   | www.necen.nl
m.maletta at nki.nl
Tel : +31 20 512 2022
Fax : +31 20 512 2029
________________________________________
From: 3dem-bounces at ncmir.ucsd.edu [3dem-bounces at ncmir.ucsd.edu] On Behalf Of Sjors Scheres [scheres at mrc-lmb.cam.ac.uk]
Sent: Tuesday, 14 January 2014 11:08 AM
To: 3dem at ncmir.ucsd.edu
Subject: Re: [3dem] electron dose & data collection strategy for Titan Krios, 300kV, Falcon back thinned

One other observation that may be relevant to your experiment: with the
settings just described, the first 1-2 movie frames (out of a total of
17 for the 1s exposure) are often significantly worse than frame 3. If
you will be limited by FEI's 7-frame capturing system, you may want to
skip the first 1 or 2 frames.
HTH, S



On 01/13/2014 03:50 PM, Shabih Shakeel wrote:
>
> Dear All,
>
> I am going to collect single particle data on Titan Krios operated at
> 300kV with Falcon II (back-thinned) detector. The sample is
> icosahedral virus of 30nm diameter. Does any one have experience
> collecting data in these settings?
>
> We are especially looking for comments or advice on electron dosage to
> use and number of frames to record with an aimed resolution of 3-5Å.
> This is our one shot at collecting the data for this sample, so we
> really want it to work.
>
> Of course, any other useful comments, advice, tips would also be
> appreciated.
>
> Thanks a lot.
>
> Best Regards,
> Shabih Shakeel
>
>
> ---------------------------------------------
> Shabih Shakeel, MSc
> Doctoral researcher
> Butcher group
> Institute of Biotechnology
> P.O. Box 65 (Viikinkaari 1)
> 00014 University of Helsinki
> Finland
>
> cell: +358 46 8430774
>
>
> _______________________________________________
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> 3dem at ncmir.ucsd.edu
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--
Sjors Scheres
MRC Laboratory of Molecular Biology
Francis Crick Avenue, Cambridge Biomedical Campus
Cambridge CB2 0QH, U.K.
tel: +44 (0)1223 267061
http://www2.mrc-lmb.cam.ac.uk/groups/scheres

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