title: Visualization of Cellular Components in a Mammalian Cell with Liquid-Cell Transmission Electron Microscopy.
abstract: 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 (~2.5 e-/nm2 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.
authors.affiliation: 1Chemistry Department,Institute for Biochemistry and Molecular Biology,University of Hamburg,Martin-Luther-King Platz 6,20146 Hamburg,Germany.
authors.forename: Stephanie
authors.identifier: 0000-0001-7382-1799
authors.initials: S
authors.lastname: Besztejan
authors.affiliation: 3Max Planck Institute for the Structure and Dynamics of Matter,Luruper Chaussee 149,Geb. 99 (CFEL),22761 Hamburg,Germany.
authors.forename: Sercan
authors.initials: S
authors.lastname: Keskin
authors.affiliation: 3Max Planck Institute for the Structure and Dynamics of Matter,Luruper Chaussee 149,Geb. 99 (CFEL),22761 Hamburg,Germany.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Manz
authors.affiliation: 3Max Planck Institute for the Structure and Dynamics of Matter,Luruper Chaussee 149,Geb. 99 (CFEL),22761 Hamburg,Germany.
authors.forename: Günther
authors.initials: G
authors.lastname: Kassier
authors.affiliation: 3Max Planck Institute for the Structure and Dynamics of Matter,Luruper Chaussee 149,Geb. 99 (CFEL),22761 Hamburg,Germany.
authors.forename: Robert
authors.initials: R
authors.lastname: Bücker
authors.affiliation: 4Institute of Microsystems Technology,Hamburg University of Technology (TUHH),Eißendorfer Straße 42,21073 Hamburg,Germany.
authors.forename: Deybith
authors.initials: D
authors.lastname: Venegas-Rojas
authors.affiliation: 4Institute of Microsystems Technology,Hamburg University of Technology (TUHH),Eißendorfer Straße 42,21073 Hamburg,Germany.
authors.forename: Hoc K
authors.initials: HK
authors.lastname: Trieu
authors.affiliation: 5Institute of Biochemistry,Westfälische Wilhelms-Universität Münster,Wilhelm-Klemm-Strasse 2,48149 Muenster,Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Rentmeister
authors.affiliation: 2The Hamburg Centre for Ultrafast Imaging,University of Hamburg, Luruper Chaussee 149,22761 Hamburg,Germany.
authors.forename: R J Dwayne
authors.initials: RJ
authors.lastname: Miller
mesh_terms: D000954:Antigens, Surface; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor / Q000648:ultrastructure; D004583:Electrons; D043425:Glutamate Carboxypeptidase II; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008297:Male; D044085:Microfluidics / Q000379:methods*; D046529:Microscopy, Electron, Transmission / Q000379:methods*; D055616:Microtechnology / Q000379:methods; D053758:Nanoparticles / Q000737:chemistry*; D011471:Prostatic Neoplasms; D017655:Silicon Compounds; D013194:Staining and Labeling
chemical_list: D000954:Antigens, Surface; D017655:Silicon Compounds; C473848:FOLH1 protein, human; D043425:Glutamate Carboxypeptidase II; C032734:silicon nitride
keywords: 
                 in situ imaging; Liquid-cell TEM; gold nanoparticles; mammalian cells; nanofluidic cell