Dear Bum and Takanori,
El 12/12/2018 a las 9:02, Takanori Nakane escribió:
> Hi,
>
> The advantage of non-gain-corrected data is that it is easier to
> compress.
I totally agree with this.
> The quality of motion correction is the same.
I would disagree with this statement. Depending on the range of the gain
image (case 1: from 0.999 to 1.001; or case 2: from 0.5 to 2), the movie
alignment may be driven by the particles in the image (case 1) or by the
gain image (case 2, in this case I would expect that the alignment
algorithm does not want to move the images at all because it would mean
disaligning the gain images which have a lot of energy).
My recommendation would be keep the uncorrected (compressed if you want)
images for storage efficiency, and always correct for the gain before
doing any operation with those movies (the corrected version can be
deleted after finishing the operation so that it does not take space).
Kind regards, Carlos Oscar
>
> Unfortunately, you cannot save non-gain corrected data from Falcon.
> It is available only for K2/K3.
>
> Best regards,
>
> Takanori Nakane
>
> On 2018/12/12 2:58, 류범한 wrote:
>> Hello, CCPEM.
>>
>> So far I have always collected gain-corrected data with EPU (& Titan
>> Krios+Falcon3 / FEI).
>>
>> But in somewhere I read that ‘non-gain-corrected data' would be
>> better in motion tracking with separately prepared gain reference
>> image. So I am about to check how much it would be helpful, but I
>> never tried this before.
>>
>> Regarding this, I have several questions.
>>
>>
>> 1. What is the actual advantage of ’non-gain-corrected data’? in what
>> terms exactly?
>> (Simply tracking algorithms of motion correction softwares are
>> slightly better than EPU?)
>>
>> 2. In practice, how much is it advantageous? in what terms?
>> (Especially in the aspect of final resolution of refined 3D map)
>>
>> 3. How can I collect non-gain-corrected data ?
>> By selecting ‘Bias’ option of Bias/Gain correction setting in CCD/TV
>> camera TAB on TEM user interface?
>>
>> 4. How can I prepare gain reference separately and correctly?
>> - Simply take from Falcon Reference manager? But file extension is
>> .RAW other than .mrc. :(
>> - Set the imaging condition exactly same with that of data
>> collection, and just take gain reference image w/o sample or in empty
>> hole area? (then check it via TIA?)
>>
>>
>> Thanks in advance.
>>
>>
>>
>> Bum Han Ryu
>>
>> Postdoctoral Researcher
>> Korea Basic Science Institute (KBSI)
>> 161 Yeongudanji-ro, Ochang-eup, Cheongwon-gu, Cheongju-si
>> Chungcheongbuk-do, Republic of Korea
>>
>> Office) +82-43-240-5329
>> Mobile) +82-10-7119-0505
>>
>>
>>
>>
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>
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Carlos Oscar Sánchez Sorzano e-mail: [log in to unmask]
Biocomputing unit http://i2pc.es/coss
National Center of Biotechnology (CSIC)
c/Darwin, 3
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