By Thomas Harry Sharp
As a part of a collaboration among assorted teams in chemistry and biochemistry, Thom Sharp offers the following his thesis paintings at the improvement of recent tools for cryoelectron microscopy. all through his Ph.D., Thom needed to grasp an entire variety of suggestions together with modelling, molecular biology and microscopy. utilizing those talents to take on an exceptional challenge, the pursuit of high-resolution buildings of peptide-based fabrics, Thom highlights during this thesis his newly built tools for analysing and processing this actual form of electron microscopy information. This thesis provides the 1st molecular description of a de-novo designed peptide-based fabric. more often than not, this learn could have a big impact at the peptide meeting box, and likewise in electron microscopy because it introduces new tools and methods, all of that are Thom's innovations and are defined during this thesis.
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Extra resources for Biomolecular Imaging at High Spatial and Temporal Resolution In Vitro and In Vivo
Pure water can be cooled down to −38 ◦ C and remain liquid, so-called supercooling. However, as soon as this homogenous nucleation temperature is reached every water molecule can behave as a crystal nucleus and the water rapidly freezes to form hexagonal ice (the familiar species found on mountains and in freezers) . As soon as crystallisation begins the latent heat of fusion released by crystallisation increases the temperature until crystallisation is complete. The only way to avoid hexagonal ice formation is to cool the water extremely quickly forming so-called vitreous ice, which due to this difficulty was discovered only 30 years ago .
To α and β to β) cease and α to β contacts occur. This was thought to be a mechanism of depolymerisation, where microtubules peeled apart along the seam. CryoTEM, however, allowed Sui et al. to image microtubules with varying numbers of protofilaments with differing helix start-numbers, which has shown that this is not the case . Sui et al. had access to a 400 kV cryoTEM which has a better resolution than 200 kV TEMs used for imaging actin by virtue of the higher accelerating voltage .
82] and the 6 Å map from (a) (b) 3 4 (c) 2 1 Fig. 16 The structures of G- and F-actin. a G-actin (PDB entry 1J6Z) is subdivided into 2 lobes, each with 2 subdomains (1–4 as numbered) with one molecule of ADP between them (shown as sticks) . 6 Å  c (EMDB entry 5168). 4 Protein and Peptide Fibres 25 Murakami et al.  (Fig. 16). Key to their success was a TEM equipped with a FEG combined with in-column energy filter (see Sect. 2). This increased the visibility of the otherwise low contrast 6 nm wide filaments.