Visualization of Cellular Components in a Mammalian Cell with Liquid-Cell Transmission Electron Microscopy

被引：9

作者：

Besztejan, Stephanie ^{[1
,2
]}

Keskin, Sercan ^{[3
]}

Manz, Stephanie ^{[3
]}

Kassier, Guenther ^{[3
]}

Buecker, Robert ^{[3
]}

Venegas-Rojas, Deybith ^{[4
]}

Trieu, Hoc K. ^{[4
]}

Rentmeister, Andrea ^{[5
]}

Miller, R. J. Dwayne ^{[2
,3
,6
,7
]}

机构：

[1] Univ Hamburg, Inst Biochem & Mol Biol, Dept Chem, Martin Luther King Pl 6, D-20146 Hamburg, Germany

[2] Univ Hamburg, Hamburg Ctr Ultrafast Imaging, Luruper Chaussee 149, D-22761 Hamburg, Germany

[3] Max Planck Inst Struct & Dynam Matter, Luruper Chaussee 149,Geb 99 CFEL, D-22761 Hamburg, Germany

[4] Hamburg Univ Technol TUHH, Inst Microsyst Technol, Eissendorfer Str 42, D-21073 Hamburg, Germany

[5] Wilhelms Univ Munster, Inst Biochem, Wilhelm Klemm Str 2, D-48149 Munster, Germany

[6] Univ Toronto, Dept Chem, 80 St George St, Toronto, ON M5S3H6, Canada

[7] Univ Toronto, Dept Phys, 80 St George St, Toronto, ON M5S3H6, Canada

来源：

MICROSCOPY AND MICROANALYSIS | 2017年 / 23卷 / 01期

关键词：

Liquid-cell TEM; mammalian cells; nanofluidic cell; in situ imaging; gold nanoparticles; BIOLOGICAL SAMPLES; GOLD NANOPARTICLES; NUCLEAR-ENVELOPE; MEMBRANE ANTIGEN; SPECIMEN DAMAGE; RNA MOLECULES; WHOLE CELLS; LIVE CELLS; RESOLUTION; DYNAMICS;

D O I：

10.1017/S1431927616012708

中图分类号：

T [工业技术];

学科分类号：

08 ;

摘要：

We present liquid-cell transmission electron microscopy (liquid-cell TEM) imaging of fixed and non-fixed prostate cancer cells (PC3 and LNCaP) with high resolution in a custom developed silicon nitride liquid cell. Fixed PC3 cells were imaged for 90-120 min without any discernable damage. High contrast on the cellular structures was obtained even at low electron doses (similar to 2.5 e(-)/nm(2) per image). The images show distinct structures of cell compartments (nuclei and nucleoli) and cell boundaries without any further sample embedding, dehydration, or staining. Furthermore, we observed dynamics of vesicles trafficking from the cell membrane in consecutive still frames in a non-fixed cell. Our findings show that liquid-cell TEM, operated at low electron dose, is an excellent tool to investigate dynamic events in non-fixed cells with enough spatial resolution (few nm) and natural amplitude contrast to follow key intracellular processes.

引用

页码：46 / 55

页数：10

共 59 条

[1] IMIDAZOLE-BUFFERED OSMIUM-TETROXIDE - AN EXCELLENT STAIN FOR VISUALIZATION OF LIPIDS IN TRANSMISSION ELECTRON-MICROSCOPY [J].