By Joachim Frank
This concise and straightforward to learn ebook is essentialy a crash path in unmarried particle reconstruction via electron microscopy. it is not precisely a how-to guide, extra an summary of ways the total procedure works. hence, it really is worthwhile not just to somebody who desires to how to do that, but in addition to those that will be drawn to identifying if this system will be important to them. and really, the full subject is satisfactorily cool that i'd suggest the ebook even to those that understand they'll by no means use this system, simply because its enjoyable to examine it. The ebook is obviously written and does not suppose alot of previous wisdom. A easy undergrad schooling in math will be sufficient to address the equations.
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1982). Nevertheless, negative staining has been used with great success in numerous computer reconstructions of viruses and other large regular macromolecular assemblies. The reason such reconstructions are legitimate is that they utilize symmetries which allow many views to be generated from the view least affected by the distortion. " As this book shows, the problems in the approach of Hoppe et al. to the reconstruction of negatively stained molecules can be overcome by the use of a radically different method of data collection.
B. Negative Staining: Principle The negative staining method, which goes back to Brenner and Horne (1959), has been widely used to obtain images of macromolecules with high contrast. Typically, an aqueous suspension is mixed with 1 to 2% uranyl acetate and applied to a carbon-coated copper grid. The excess liquid is blotted away and the suspension is allowed to dry. Although, to some extent, the stain goes into aqueous channels, the structural information in the image is basically limited to the shape of the molecule as it appears in projection.
So even if not strictly correct, the colloquial term "power spectrum" is not too much off the mark. Another note concerns the display mode. , the square root of the power spectrum, because of the limited dynamic range of monitor screens. Logarithmic displays are also occasionally used, but experience shows that these often lead to an unacceptable compression of the dynamic range, rendering zeros of the CTF virtually invisible. F. Determination of the Contrast Transfer Function The CTF may either be determined "by hand," specifically by measuring the positions of the zeros and fitting them to a chart of the CTF characteristics, or by using automated computer-fitting methods.