RETRACTED: Liquid-EM goes viral – visualizing structure and dynamics

Kelly, Deborah F.; DiCecco, Liza‐Anastasia; Jonaid, G. M.; Dearnaley, W. J.; Spilman, Michael; Gray, Jennifer L.; Dressel-Dukes, Madeline J.

doi:10.1016/j.sbi.2022.102426

Cited by 13 publications

(10 citation statements)

References 60 publications

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“…Liquid-cell electron microscopy (LCEM) [1][2][3][4][5][6][7] is a known state-of-the-art technique used to perform high-magnification imaging of nanomaterials and biological specimens which are suspended in a liquid inside of a nanofluidic cell (NFC). There are two types of LCEM systems; those connected to a syringe pump and capable of conferring liquid flow, commonly known as 'flow LCEM systems' [8][9][10][11][12][13][14][15][16][17][18][19][20][21][22][23][24] , and those which are stationary systems [25][26][27][28][29][30][31][32][33][34][35][36][37][38][39][40][41][42] herein referred to as 'static LCEM systems'.…”

Section: Mainmentioning

confidence: 99%

High-Resolution Bulgeless Liquid-Cell Electron Microscopy

Lott

Petruk

Shaw

et al. 2023

Preprint

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Liquid cell electron microscopy (LCEM) has long suffered from irreproducibility and its inability to confer high-quality images over a wide field of view. LCEM demands the encapsulation of the in-liquid sample between two ultrathin membranes (windows). In the vacuum environment of the electron microscope, the windows bulge, drastically reducing the achievable resolution and the usable viewing region. Herein, we introduce a shape-engineered nanofluidic cell architecture and an air-free drop-casting sample loading technique, which combined, provide robust bulgeless imaging conditions. We demonstrate the capabilities of our approach through the study of in-liquid model samples and quantitative measurements of the liquid layer thickness. The presented LCEM method confers high throughput, lattice resolution across the complete viewing window, and sufficient contrast for the observation of unstained liposomes, paving the way to high-resolution movies of biospecimens in their near native environment.

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Section: Mainmentioning

confidence: 99%

High-Resolution Bulgeless Liquid-Cell Electron Microscopy

Lott

Petruk

Shaw

et al. 2023

Preprint

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“…Liquid TEM, the room temperature correlate to cryo TEM, is a relatively new imaging technique that can provide HR and real-time information on hydrated biological states. [33][34][35][36][37][38][39][40][41] While cryo TEM routinely provides HR imaging and is considered the gold standard for biological imaging, recent technical breakthroughs in acquisition strategies and thin film enclosures are pushing liquid TEM imaging resolution to comparable levels with the additional benefit of retaining the liquid state. 40 Liquid imaging is often performed in one of two ways: with a commercial liquid TEM holder or by encapsulation methods.…”

Section: Liquid Transmission Electron Microscopy For Probing Collagen...mentioning

confidence: 99%

Liquid Transmission Electron Microscopy for Probing Collagen Biomineralization

DiCecco

Gao

Gray

et al. 2023

Preprint

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Collagen biomineralization is foundational to hard tissue assembly. While studied extensively, collagen mineralization processes are not fully understood as the majority of theories are derived from electron microscopy (EM) in static, dehydrated, or frozen conditions, unlike the liquid phase environment where mineralization occurs dynamically. Herein, novel liquid transmission EM (TEM) strategies are presented, where collagen mineralization was explored in liquid conditions for the first time. Custom thin-film enclosures were employed to visualize the mineralization of reconstituted collagen fibrils in a calcium-phosphate and polyaspartic acid solution to promote intrafibrillar mineralization. TEM highlighted that at early time points, precursor mineral particles attached to collagen and progressed to crystalline mineral platelets aligned with fibrils at later time points. This aligns with observations from other techniques and validates this liquid TEM approach. This work provides a new liquid imaging approach for exploring collagen biomineralization, advancing toward understanding disease pathogenesis and remineralization strategies for hard tissues.

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“…Liquid-cell electron microscopy (LCEM) 1–7 is a known state-of-the-art technique used to perform high-magnification imaging of nanomaterials and biological specimens which are suspended in a liquid inside of a nanofluidic cell (NFC). There are two types of LCEM systems; those connected to a syringe pump and capable of conferring liquid flow, commonly known as ‘flow LCEM systems’, 8–24 and those which are stationary systems 25–42 herein referred to as ‘stationary LCEM systems’.…”

Section: Introductionmentioning

confidence: 99%