Cellular-, 3D- and Correlative Electron Microscopy

Dr. Yannick Schwab

Team Leader & Head, Electron Microscopy Core Facility

EMBL, Heidelberg, Germany

EMBL Australia and the Cell Motility & Mechanobiology group are proud to host Dr. Yannick

Schwab for a special seminar at 1:30-2:30pm, Thursday 20th July 2017.

Cellular-, 3D- and Correlative Electron Microscopy

Abstract Correlative light and electron microscopy (CLEM) is a set of techniques that allow

data acquisition with both imaging modalities on a single object. One common challenge when

trying to combine imaging modalities on the same sample is to identify space cues (external or

internal) to track single objects when switching from light microscopy (LM) to electron

microscopy (EM). On adherent cultured cells, we have previously developed specific substrates

with coordinates to precisely record the position of cells (Spiegelhalter et al., 2009).

On more complex specimens, such as multicellular organisms, this targeting is even more

critical, as systematic EM acquisition of their entire volume is close to impossible. For this

reason, we are developing new methods to map the region of interest (ROI) within large living

specimens, taking advantage of structural hallmarks in the sample that are visible with both LM

and EM. The position of the ROI is mapped in 3D by confocal or multiphoton microscopy and

then tracked at the EM level by targeted ultramicrotomy (Kolotuev et al. 2009; 2012; Goetz et al.

2014). Relying on structural features of the sample as anchor points, the cell or structure of

interest can then be retrieved with sub-micrometric precision (Durdu et al. 2014, Goginashvili et

al. 2015, Hampoelz et al 2016).

References

• Spiegelhalter C et al. (2010), PLoS ONE 5(2):e9014. doi: 10.1371/journal.pone.0009014

• Kolotuev I, Schwab Y, Labouesse M. (2010), Biol. Cell 102(2):121-132. doi: 10.1042/bc20090096

• Kolotuev I… Schwab Y. (2012), Methods Cell Biol. 111:203-222. doi: 10.1016/b978-0-12-416026-2.00011-x

• Goetz JG, et al. (2014), Cell Rep 6(5):799-808. doi: 10.1016/j.celrep.2014.01.032

• Durdu S, et al. (2014), Nature 515(7525):120-124. doi: 10.1038/nature13852

• Goginashvili A, et al. (2015), Science 347(6224):878-882. doi: 10.1126/science.aaa2628

• Hampoelz B et al. (2016), Cell 166(3):664-678. doi: 10.1016/j.cell.2016.06.015