[3dem] One-sided vs two-sided blotting

[Bill & Sue Tivol]

> On Sep 14, 2017, at 8:35 PM, Masahide Kikkawa <mkikkawa at gmail.com> wrote:
> 
> One of the advantage of one-sided blotting is the “filtering” effect of the holey carbon membrane for filamentous/tubular samples. 
> In this case, you can use lower concentrations of samples, because you can “concentrate” samples on the grid.
> ---------------------------------------------------------------
> Masahide Kikkawa, M.D., Ph. D.
> Professor
> Department of Cell Biology & Anatomy
> Graduate School of Medicine
> The University of Tokyo
> http://structure.m.u-tokyo.ac.jp/ <http://structure.m.u-tokyo.ac.jp/>
> tel: +81-3-5841-3338
> fax: +81-3-5841-3339
> ---------------------------------------------------------------
>> On Sep 15, 2017, at 12:31, Moni Banerjee <banerjee.moni at gmail.com <mailto:banerjee.moni at gmail.com>> wrote:
>> Dear all,
>> 
>>  I am wondering about the advantages (or disadvantages) of one-sided vs two-sided blotting during sample freezing for SPA. Do people have preferences for one or the other depending on the type of sample? Does one-sided blotting lead to thicker ice in general? Any suggestion/comment will be very welcome.
>>  Thanks,
>>  Manidipa
>> ------------------------
>> Dr. Manidipa Banerjee, PhD
>> Associate Professor
>> School of Biological Sciences
>> Indian Institute of Technology, Delhi
>> New Delhi 110016
>> India
>> _______________________________________________
> 
Dear Mandipa & Masahide,
	One specimen I worked with had a much greater affinity for the blot paper than for the grid, so I had not only to try one-sided blotting, but I ended having to blot from the side of the grid to have the specimen stay on.  This was a difficult process to get consistent ice thickness—mostly I failed—and it is a numbers game to get good enough specimens to get anything of value.  One can get sufficiently thin ice from either one-sided or edge blotting, but it requires more skill.  As usual, specimen preparation is the key to microscopy, and it is definitely specimen dependent.
						Yours,
						Bill



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