Super-resolution and/or electron microscopy approaches may provide more insight into how actin filament structure and/or turnover is altered in the absence of palmitoyl-LIMK1.
By directly labeling impβ with a fluorescent reporter, we removed additional localization uncertainty error (commonly referred to as linkage error) associated with the antibody labeling methods normally used for super-resolution or electron microscopy.
Utilizing a combination of super-resolution, light, and electron microscopy, we revealed that nanotubes are composed of chains of membranous segments harboring a continuous lumen.
Either way, advanced microscopy approaches (live-cell imaging, super-resolution microscopy and electron microscopy/tomography) in combination with computational modeling have now provided strong evidence that the processing/remodeling of ring-like ESCRT-III filaments by Vps4 is a prerequisite for membrane scission in cytokinesis.
Besides super-resolution fluorescence microscopy, conventional electron microscopy (EM) combined with immunolabeling or clonable EM tags provides a unique approach to correlate protein localization information and cellular ultrastructural information.
Recent technological advances in quantitative and cross-linking mass spectrometry, super-resolution fluorescence microscopy and electron microscopy have enormously accelerated the integration of different types of data into coherent structural models.
In contrast to most fluorophores, however, silver and gold nanoparticles do not photobleach and have higher extinction coefficients they can also be imaged with super-resolution microscopy methods and electron beams that surpass the ca. 200 nm diffraction resolution limit of a conventional LSCM (Zhang et al., 2015).
Common human hematopoietic progenitors, myeloid precursor cells, differentiated monocytes and granulocytes analyzed by super-resolution fluorescence microscopy and electron microscopy revealed profound differences with respect to global chromatin arrangements, the nuclear space occupied by the interchromatin compartment and the distribution of nuclear pores.
Research in this field urgently calls for advanced experimental systems and tools such as single-molecule recording, in situ electron cryotomography, super-resolution microscopy, quantitative cross-linking MS, as well as other new biochemical/biophysical methods, pharmacological agents, antibodies, and animal models (Pack et al., 2014; Asano et al., 2015; Lu et al., 2015).
