Correlative light and electron microscopy: from live cell dynamic to 3D ultrastructure

Coralie Spiegelhalter (1, 2) , Jocelyn F. Laporte (1, 2) , Yannick Schwab

1 IGBMC - Institut de Génétique et de Biologie Moléculaire et Cellulaire
2 IGBMC - Institut de génétique et biologie moléculaire et cellulaire

Correlative light and electron microscopy (CLEM) aims at combining data acquired from the same sample through both imaging modalities. Many combinations can be found in the literature where almost any kind of light microscopy (LM) has been associated to different processing in electron microscopy (EM) and applied to a wide variety of specimen, from cultured cells to multicellular organisms. In this chapter, we focus on a technique that intends to combine LM acquisition on living cells with transmission EM (TEM) analysis. A specific attention is given to the description of a method to bring precise coordinates to the object of interest, to allow a straightforward correlation between LM and EM. Moreover, we describe how, by using high-pressure freezing as a fixation technique, dynamic events observed at the LM are captured and studied at the ultrastructural level.

Domaines: Biochimie, Biologie Moléculaire

Coralie Spiegelhalter, Jocelyn F. Laporte, Yannick Schwab. Correlative light and electron microscopy: from live cell dynamic to 3D ultrastructure. John Kuo. Methods in Molecular Biology, 1117, Humana Press, pp.485-501, 2013, 1064-3745, 978-1-62703-776-1. ⟨10.1007/978-1-62703-776-1_21⟩. ⟨hal-04383932⟩