The CryoCapsule: Simplifying Correlative Light to Electron Microscopy

被引:25
|
作者
Heiligenstein, Xavier [1 ,2 ]
Heiligenstein, Jerome [3 ,4 ]
Delevoye, Cedric [1 ,2 ]
Hurbain, Ilse [1 ,2 ,5 ]
Bardin, Sabine [1 ,6 ]
Paul-Gilloteaux, Perrine [1 ,5 ,7 ]
Sengmanivong, Lucie [1 ,5 ,7 ]
Regnier, Gilles [3 ]
Salamero, Jean [1 ,5 ,7 ]
Antony, Claude [8 ]
Raposo, Graca [1 ,2 ,5 ]
机构
[1] Inst Curie, Ctr Rech, F-75248 Paris, France
[2] CNRS, Struct & Membrane Compartments, UMR144, F-75248 Paris, France
[3] CER Paris, Arts & Metiers ParisTech, CNRS, UMR 8006, Paris, France
[4] CryoCapCell, F-75015 Paris, France
[5] Cell & Tissue Imaging Facil PICT IBiSA, CNRS, UMR144, F-75248 Paris, France
[6] CNRS, Mol Mech Intracellular Transport, UMR144, F-75248 Paris, France
[7] CNRS, Spatio Temporal Modeling Imaging & Cellular Dynam, UMR144, F-75248 Paris, France
[8] INSERM, CNRS, Inst Genet & Biol Mol & Cellulaire, Dept Struct Biol & Genom,UMR7104,U964, Illkirch Graffenstaden, France
关键词
correlative light to electron microscopy; CryoCapsule; endosomal network; freeze substitution; high-pressure freezing; image registration; langerin; melanosomes; spatial resolution; temporal resolution; Xenopus laevis mitotic spindle; CRYOELECTRON MICROSCOPY; CELL; FLUORESCENCE; VISUALIZATION; CRYOFIXATION;
D O I
10.1111/tra.12164
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Correlating complementary multiple scale images of the same object is a straightforward means to decipher biological processes. Light microscopy and electron microscopy are the most commonly used imaging techniques, yet despite their complementarity, the experimental procedures available to correlate them are technically complex. We designed and manufactured a new device adapted to many biological specimens, the CryoCapsule, that simplifies the multiple sample preparation steps, which at present separate live cell fluorescence imaging from contextual high-resolution electron microscopy, thus opening new strategies for full correlative light to electron microscopy. We tested the biological application of this highly optimized tool on three different specimens: the in vitro Xenopus laevis mitotic spindle, melanoma cells over-expressing YFP-langerin sequestered in organized membranous subcellular organelles and a pigmented melanocytic cell in which the endosomal system was labeled with internalized fluorescent transferrin.
引用
收藏
页码:700 / 716
页数:17
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