Applications Leica EM PACT2

CLEM_reSOLUTION_Extract_07_en.pdf

Poster_CLEM_02.pdf

"Combining live imaging with high resolution electron microscopy is a real challenge!"

“With the introduction of Green Fluorescent Protein (GFP) technology, cell biology and life sciences in general have entered a whole new exciting era of research. [...] In some instances however, the resolution of the light microscope is the limiting factor in answering our scientific questions. In these cases, the higher resolution of the electron microscope is essential. Combining both light and electron microscopy is my field of interest.

By performing so-called Correlative Light Electron Microscopy (CLEM) experiments one has the advantage of live cell imaging in the confocal microscope and afterwards have high resolution results from the transmission electron microscope of the same cell.

The Leica EM RTS was specifically developed to be used in such experiments in conjunction with Leica EM PACT2. It provides a high time resolution between the light and electron microscope, allowing excellent preservation of the ultrastructure close to the natural state, an essential prerequisite for electron microscopy. It allows us to decide upon the exact moment of interest and study that particular event at high resolution.”

Dr. Paul Verkade, Max Planck Institute for Molecular Cell Biology and Genetics, Dresden, Germany Dr. Verkade works with the Leica EM PACT2 & EM RTS High Pressure Freezer.

Piece of cartilage from the hip of a mouse. Bar represents: 10 .m.

Piece of cartilage from the hip of a mouse. Bar represents: 5 .m.

Piece of cartilage from the hip of a mouse. Bar represents: 500nm

Piece of cartilage from the hip of a mouse were cryofixed with the Leica EM PACT and freeze-substituted in acetone containing 0,5 % glutaraldehyde and 0.2 % uranyl acetate and low-temperature embedded in Lowicryl HM20. Sections were labelled for -Cop, detected with protein A gold 10 nm.

Courtesy of:
E.G. van Donselaar, B.M. Humbel
Electron Microscopy & Structure Analysis, Department of Molecular Cell biology, Utrecht University, The Netherlands

J.W. Slot
Department of Cell Biology, University Medical Center Utrecht, The Netherlands