Cell-free extract systems and the cytoskeleton: preparation of biochemical experiments for transmission electron microscopy.
Cell-free systems can be used to reconstitute complex actin or microtubule-based phenomena. For example, a number of extracts support actin-dependent propulsion of Listeria monocytogenes, whereas Xenopus laevis extracts support formation of a microtubule-based meiotic spindle. Working in vitro opens these complex processes to biochemical dissection. Here, we describe methods to view these in vitro preparations by thin-section electron microscopy.
Department of Systems Biology, Harvard Medical School, Boston, MA, USA.
Margaret
M
Coughlin
William M
WM
Brieher
Ryoma
R
Ohi
D000199:Actins / Q000378:metabolism; D000818:Animals; D001710:Biotin; D002474:Cell-Free System; D003599:Cytoskeleton / Q000378:metabolism* / Q000648:ultrastructure*; D005260:Female; D005404:Fixatives; D008089:Listeria monocytogenes / Q000378:metabolism / Q000648:ultrastructure; D046529:Microscopy, Electron, Transmission / Q000379:methods*; D010063:Ovum / Q000378:metabolism / Q000648:ultrastructure; D016613:Plastic Embedding; D008941:Spindle Apparatus / Q000378:metabolism / Q000648:ultrastructure; D019809:Streptavidin; D014982:Xenopus laevis
D000199:Actins; D005404:Fixatives; D001710:Biotin; D019809:Streptavidin