Cryogenic FIB Lift-out as a Preparation Method for Damage-Free Soft Matter TEM Imaging

Parmenter, Christopher; Fay, Michael W.; Hartfield, Cheryl; Amador, Gonzalo; Moldovan, Grigore

doi:10.1017/s1431927614007855

Cited by 7 publications

(9 citation statements)

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“…Early published work delivered cryogenic-Lift-Out (cryo-LO) on a lamella mounted to a copper support post using platinum deposition (Rubino et al, 2012) or water from the Selective Carbon Mill GIS (Gas Injection System) (Parmenter et al, 2014), through a combination of hardware modifications Fig. 1.…”

Section: Introductionmentioning

confidence: 99%

“…Successful transfer and imaging in a cryo-TEM was viewed as a significant achievement and that cryo-LO had become a potential option. Since that work, the technique has been demonstrated to work for different samples including spores, hydrogels, yeast and tissue (Rubino et al, 2012;Parmenter et al, 2014;Mahamid et al, 2015;Schaffer et al, 2019). The cryo-LO approach can of course be applied to any sample in the FIB-SEM, subject to the cooled stage and manipulator being present and this has been evidenced in the case of hard-soft interfaces such as battery materials (Zachman et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

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Cryo‐FIB‐lift‐out: practically impossible to practical reality

Parmenter

Nizamudeen

2020

Journal of Microscopy

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In this paper, we explore the development of the Cryo-Lift-Out (cryo-LO) technique as preparation tool for cryogenic transmission electron microscopy (cryo-TEM). What started in early work defying what was considered 'practically impossible' has developed into state-of-the-art practical reality. This paper presents the key hardware, basic principles and key considerations for the practical usage of cryogenic Lift-Out for those new to the field. Detailed protocols and in-depth description of considerations and points for further development are presented. The authors have attempted to formalise everything known about the technique gathered together from their expertise gained in the development of this approach.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Cryo‐FIB‐lift‐out: practically impossible to practical reality

Parmenter

Nizamudeen

2020

Journal of Microscopy

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“…[52] These results suggest that by using cryo-FIB methods, foils of organic thin films could be prepared in which the electronic structure is retained, and full implementation of the commonly used lift-out technique is starting to be employed under cryo conditions. [53] For the interfaces present in OPVs, sample preparation using a FIB in combination with damage reduction techniques, demonstrate that TEM samples could be prepared for analysis of the electronic structure at interfaces.…”

Section: Figure 4 A) Cross-sectional Hrtem Image From a Cupc/c60 Bi-lmentioning

confidence: 99%

Revealing structure and electronic properties at organic interfaces using TEM

Gilchrist

Heutz

McComb

2017

Current Opinion in Solid State and Materials Science

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Localization of Subsurface Structures for Site-Specific Cryo-FIB Lift-Out Preparation of Solid-Liquid Interfaces

Zachman

Kourkoutis

2016

Microsc Microanal

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While proof of concept demonstrations of cryo-FIB lift-out have recently been discussed [1][2][3][4][5], a method of localizing subsurface structures that can guide the TEM sample preparation of non-biological systems is thus far absent. Analogous to FIB lift-out routinely used to prepare electron transparent TEM samples from bulk materials, cryo-FIB lift-out promises the ability to extract structures in their near-native environment from large intact soft, liquid, or hard-soft materials. However, the limited throughput of the technique and small volume of material prepared require localization of buried structures of interest prior to milling. This issue was recently addressed in biology, where fluorescent labeling of molecules and cryo-fluorescence light microscopy (cryo-FLM) can be used to localize structures prior to cryo-FIB preparation [5]. Cryo-FIB and cryo-FLM images can then be correlated to identify regions of interest for milling. This technique cannot be applied to samples that lack fluorescent labels, however, so an alternative localization technique is needed for these systems.We present here a technique for localizing subsurface structures in intact hard-soft materials systems, at optimal depths for cryo-FIB lift-out. Both backscattered electron (BSE) imaging and energy dispersive x-ray spectroscopy (EDX) mapping can probe structures below the specimen's surface. A model system of iron oxide particles grown in a silica hydrogel was used to evaluate the capabilities of these signals. Electron scattering calculations revealed the characteristic Fe-Kα x-ray generation depth exceeds the range of BSE, as depicted in Figure 1a. Monte Carlo simulations were then used to directly compare BSE and EDX signals from an appropriately sized iron oxide particle embedded at various depths in water under a 10 nm platinum layer, scanned with a 30 kV electron beam, as shown in Figure 1b. These results confirm, while BSE imaging can identify heavy particles near the surface, EDX mapping can readily localize particles with element specificity to depths beyond the typical size of a lamella. These results are consistent with experimental observations in the cryo-FIB, shown in Figure 2. EDX was used to localize individual iron oxide particles at the proper depth in our hydrogel system and guide precision milling. Thin and homogeneous lamellas were then prepared for cryo-STEM, as shown in Figure 2c, enabling high-resolution cryo-STEM imaging to be performed on the particles, as in Figure 2f. EDX has thus proven to be an effective tool for localizing subsurface structures in hard-soft materials for cryo-FIB lift-out and subsequent high-resolution characterization by cryo-STEM. Consequently, use of this technique can dramatically increase the yield of desired structures in lamellas of appropriate samples, significantly increasing the effectiveness of cryo-FIB lift-out for a number of fields [6].[1] S.

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Cryogenic FIB Lift-out as a Preparation Method for Damage-Free Soft Matter TEM Imaging

Cited by 7 publications

References 0 publications

Cryo‐FIB‐lift‐out: practically impossible to practical reality

Cryo‐FIB‐lift‐out: practically impossible to practical reality

Revealing structure and electronic properties at organic interfaces using TEM

Localization of Subsurface Structures for Site-Specific Cryo-FIB Lift-Out Preparation of Solid-Liquid Interfaces

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