[3dem] Tomography to visualize interaction of liposomes

[Sonali Dhindwal]

?Dear all, Thank you for the suggestions.

As mentioned by Schmid, may be protein is getting attracted to air-water interface. Just to mention, grid I am using has an ultra-thin layer of carbon on the top, if it could also add to one of the reasons.
Misha: may be thin ice can help to bring liposomes and protein closer. Thanks.
Thanks for the reference to the paper Gary.?



--
Sonali Dhindwal
Post Doctoral Fellow
City College of New York
New York

"The dream is not that you see in sleep, dream is which does not let you sleep."?


________________________________
From: 3dem <3dem-bounces at ncmir.ucsd.edu> on behalf of Sonali Dhindwal <sdhindwal at ccny.cuny.edu>
Sent: Thursday, August 10, 2017 10:56 PM
To: 3dem at ncmir.ucsd.edu
Subject: [3dem] Tomography to visualize interaction of liposomes

?Dear 3DEM group members,

I have been studying the interaction of my protein of interest (~900 KDa and ~200Å in size) with liposomes (~2000Å is size).
Biochemical assays demonstrates their interaction and so is the images collected by negative stain and Cryo-EM.
To clearly visualize interaction, I have collected few tomograms. Though, after processing the tomograms and going through the slices, liposomes and protein seem to be in different layers.
In initial slices, I can see all the liposomes and in the later slices I see all the protein molecules. And going through the slices, first I see liposomes and as they start to fade away, I start to see the protein molecules and few protein molecules lying very close to the liposomes.
Is this normal ?
I am looking for any suggestions or advice that can be done either during processing of data or preparation of grids  that can help to improve the visualization of interactions among these molecules.
Thanks and Regards

--
Sonali Dhindwal
Post Doc

"The dream is not that you see in sleep, dream is which does not let you sleep."?

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