Correlated optical and isotopic nanoscopy.
The isotopic composition of different materials can be imaged by secondary ion mass spectrometry. In biology, this method is mainly used to study cellular metabolism and turnover, by pulsing the cells with marker molecules such as amino acids labelled with stable isotopes ((15)N, (13)C). The incorporation of the markers is then imaged with a lateral resolution that can surpass 100 nm. However, secondary ion mass spectrometry cannot identify specific subcellular structures like organelles, and needs to be correlated with a second technique, such as fluorescence imaging. Here, we present a method based on stimulated emission depletion microscopy that provides correlated optical and isotopic nanoscopy (COIN) images. We use this approach to study the protein turnover in different organelles from cultured hippocampal neurons. Correlated optical and isotopic nanoscopy can be applied to a variety of biological samples, and should therefore enable the investigation of the isotopic composition of many organelles and subcellular structures.
1] Department of Neuro- and Sensory Physiology, University of Göttingen Medical Centre, and Centre for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB), Grisebachstr. 5, 37077 Göttingen, Germany [2] International Max Planck Research School, 37077 Göttingen, Germany.
Sinem K
SK
Saka
Leibniz-Institute for Baltic Sea Research, Seestr. 15, 18119 Rostock, Germany.
Angela
A
Vogts
Department of Neuro- and Sensory Physiology, University of Göttingen Medical Centre, and Centre for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB), Grisebachstr. 5, 37077 Göttingen, Germany.
Katharina
K
Kröhnert
Cameca, 29 Quai des Grésillons, 92622 Gennevilliers, France.
François
F
Hillion
Department of Neuro- and Sensory Physiology, University of Göttingen Medical Centre, and Centre for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB), Grisebachstr. 5, 37077 Göttingen, Germany.
Silvio O
SO
Rizzoli
University of Göttingen Medical Center, Robert-Koch-Street 34, 37075 Göttingen, Germany.
Johannes T
JT
Wessels
D007554:Isotopes; D008856:Microscopy, Fluorescence / Q000379:methods*; D036103:Nanotechnology / Q000379:methods*; D018629:Spectrometry, Mass, Secondary Ion / Q000379:methods
D007554:Isotopes