Cryo-FIB specimen preparation for use in a cartridge-type cryo-TEM

Hsieh, Chyongere; Wu, Yongping; Schmelzer, Thomas; Wang, Pan; Lin, Ying; Marko, Michael; Sui, Haixin

doi:10.1016/j.jsb.2017.05.011

Cited by 15 publications

(5 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Cryo-FIB milling is simpler and requires less specialized personnel than cryo-sectioning, making it widely applied in cryo-TEM. [133][134][135] Similar protection from ion beam damage has also been explored in polymer films that suffer from beam heating due to low conductivity. The results showed enhanced tolerance to beam current by nearly 10 times.…”

Section: Innovations In Specimen Protectionmentioning

confidence: 99%

Atomic-level imaging of beam-sensitive COFs and MOFs by low-dose electron microscopy

Zhan,

Liu,

Wang

et al. 2024

Nanoscale Horiz.

View full text Add to dashboard Cite

show abstract

Section: Innovations In Specimen Protectionmentioning

confidence: 99%

Atomic-level imaging of beam-sensitive COFs and MOFs by low-dose electron microscopy

Zhan,

Liu,

Wang

et al. 2024

Nanoscale Horiz.

View full text Add to dashboard Cite

show abstract

“…The challenge of freezing hydrated Life Science samples into the vitreous state is limited by the thermal coefficient of (Strunk et al, 2012;Arnold et al, 2016;Schaffer et al, 2017;Spehner et al, 2020) Mammalian cells (Rigort et al, 2010a;Hayles et al, 2010;Chang and Joester, 2015;He et al, 2017) Yeast water itself (Gan & Jensen, 2012). As a result, the sample type and size determine the available options for preparing the samples.…”

Section: Freezing Life Science Samplesmentioning

confidence: 99%

An introduction to cryo‐FIB‐SEM cross‐sectioning of frozen, hydrated Life Science samples

Hayles¹,

Winter

2020

Journal of Microscopy

View full text Add to dashboard Cite

The introduction of cryo-techniques to the focused ion-beam scanning electron microscope (FIB-SEM) has brought new opportunities to study frozen, hydrated samples from the field of Life Sciences. Cryo-techniques have long been employed in electron microscopy. Thin electron transparent sections are produced by cryo-ultramicrotomy for observation in a cryo-transmission electron microscope (TEM). Cryo-TEM is presently reaching the imaging of macromolecular structures. In parallel, cryo-fractured surfaces from bulk materials have been investigated by cryo-SEM. Both cryo-TEM and cryo-SEM have provided a wealth of information, despite being 2D techniques. Cryo-TEM tomography does provide 3D information, but the thickness of the volume has a maximum of 200-300 nm, which limits the 3D information within the context of specific structures. FIB-milling enables imaging additional planes by creating cross-sections (e.g. cross-sectioning or site-specific X-sectioning) perpendicular to the cryo-fracture surface, thus adding a third imaging dimension to the cryo-SEM. This paper discusses how to produce suitable cryo-FIB-SEM cross-section results from frozen, hydrated Life Science samples with emphasis on 'common knowledge' and reoccurring observations.

show abstract

“…[103] These significant achievements by cryo-FIB milling demonstrate the feasibility of this technique used together with cryo-ET to prepare and study thin, distortion-free lamellae of frozen hydrated biological materials in situ. [135]…”

Section: Cryo-focused Ion Beammentioning

confidence: 99%

Cryo-ET bridges the gap between cell biology and structural biophysics

Cheng¹,

Wang²,

Shen

2018

Chinese Phys. B

View full text Add to dashboard Cite

Cryo-electron tomography (cryo-ET) is a cutting-edge technology providing three-dimensional in situ ultra-structural information of macromolecular machineries, organelles, and eukaryotic cells in their native environment at an unprecedented level of detail. Cryo-ET enables the direct observation of dynamic macromolecular architectures of bio-samples in their naturally occurring physiological state, without any harmful artifacts introduced by heavy metal staining, dehydration, and chemical fixation, which occur in traditional transmission electron microscopy. Over decades, cryo-ET has been providing insights into numerous aspects of cellular biology by revealing the pristinely preserved ultra-structures of different cellular components comprising the crowded and complex environment of the cell, thus, bridging the gap between cellular biology and structural biophysics. In this paper, we review the fundamentals of this technique, its recent advances in optics, detection devices, and computational algorithms. The enhancement of our understanding of structural cellular biology by combining these improvements, when integrated with other methods, such as cryo-focused ion beam milling, correlative light and electron microscopy, is discussed via a few examples from research groups worldwide. We also believe that cryo-ET applications in cell biology continue to provide fundamental insights into the field, revolutionizing structural biology itself.

show abstract

Cryo-FIB specimen preparation for use in a cartridge-type cryo-TEM

Cited by 15 publications

References 29 publications

Atomic-level imaging of beam-sensitive COFs and MOFs by low-dose electron microscopy

Atomic-level imaging of beam-sensitive COFs and MOFs by low-dose electron microscopy

An introduction to cryo‐FIB‐SEM cross‐sectioning of frozen, hydrated Life Science samples

Cryo-ET bridges the gap between cell biology and structural biophysics

Contact Info

Product

Resources

About