Which Negative Stain is Best for Sensitive Large Molecular Complexes in EM?

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Discussion Overview

The discussion centers around the selection of an appropriate negative stain for imaging a sensitive large molecular complex using electron microscopy (EM). The context includes considerations of ionic strength, specimen integrity, and the limitations of conducting the experiment at room temperature without a cold specimen holder.

Discussion Character

  • Debate/contested, Homework-related

Main Points Raised

  • One participant describes the need to choose a negative stain that maintains the integrity of a large molecular complex under low ionic strength conditions for electron tomography.
  • Another participant questions whether the original post is a homework question, suggesting that it may require more effort from the poster to receive help.
  • A subsequent reply reiterates the homework question concern, indicating a lack of willingness to assist without further input from the original poster.

Areas of Agreement / Disagreement

Participants do not reach a consensus on the best negative stain to use, and there is disagreement regarding the appropriateness of the original post as a homework question.

Contextual Notes

The discussion does not provide specific recommendations for negative stains or address the implications of conducting the experiment at room temperature.

i_a_n
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I have isolated a large molecular complex whose integrity is very sensitive to increasing ionic strength and thus had to be prepared for EM using low ionic strength solutions. And my goal is image this by electron tomography by collecting a dual tilt series of 120 images. Other experimental constraints are that I lack a cold specimen holder and thus must to this experiment at room temperature. From the possible negative stains that are available generally, which should I choose to prepare my specimen and why?
 
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You forgot to change the last "your" to "my". Is this a homework question?
 
Pythagorean said:
You forgot to change the last "your" to "my". Is this a homework question?
Yes. But can you help me somehow?
 
No, but someone who would be able to help you would probably appreciate some effort in answering the question on your part.
 

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