Discerning Autophagy Pathway Intermediates: Transmission Electron Microscopy Techniques

Abstract

<p>In this chapter, you will learn guidelines on how to identify macroautophagy/autophagy pathway intermediates in transmission electron microscopy samples prepared using conventional techniques including chemical fixation and plastic embedding. We will describe the typical morphological features of phagophores, autophagosomes, amphisomes and autolysosomes induced by amino-acid starvation in mammalian cells. We will point out what you should keep in mind when interpreting transmission electron microscope images taken of very thin sections cut from much thicker cells and organelles. You will also learn how immuno-electron microscopy of endogenous MAP1 LC3/LC3 can be used to help identification of autophagy pathway intermediates.<br></p>