A Brief Review of Cryobiology with Reference to Cryo Field Emission Scanning Electron Microscopy

Fleck, Roland A.; Shimoni, Eyal; Humbel, Bruno M.

doi:10.1002/9781118663233.ch11

Cited by 5 publications

(5 citation statements)

References 159 publications

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“…A comparative analysis of the pros and cons of FCS and FLIM with respect to other commonly used analytical techniques for deciphering the morphological evolution of dynamic supramolecular self-assembly is provided in Table 1. 34,40,41,46,47,59,64,78,91–106…”

Section: Discussionmentioning

confidence: 99%

Fluorescence correlation spectroscopy and fluorescence lifetime imaging microscopy for deciphering the morphological evolution of supramolecular self-assembly

2023

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Section: Discussionmentioning

confidence: 99%

Fluorescence correlation spectroscopy and fluorescence lifetime imaging microscopy for deciphering the morphological evolution of supramolecular self-assembly

2023

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“…According to an earlier report, sectioning or slicing might reduce the time duration required for cryodehydration by up to 25% [2] . Reduced sectioning or slicing of larger specimens also facilitates the study of the inner morphology of those organs [2 , 16 , 19 ]. The weight loss of different cryodehydrated specimens ranged between 60% and 80%, which is almost similar to the earlier reports [2 , 16 ].…”

Section: Discussionmentioning

confidence: 99%

“…To combat this problem, fast freezing sessions are the best way to prepare cryodehydrated specimens, as they result in smaller ice crystal formation with minimal or no tissue damage [16 , 26 ]. During the dehydration phase, it also facilitates the fluid drainage required for tissue dehydration, avoiding mentionable tissue shrinkage [16 , 17 , 19 ]. If not, larger ice crystals form inside the cells or tissues that result in the breakdown of cellular structural integrity [15 – 15 , 26 ].…”

Section: Discussionmentioning

confidence: 99%

Efficacy of cryodehydration technique in preserving the gross and histoarchitectural details of goat visceral and musculoskeletal specimens

Sultana,

Islam

2024

J Adv Vet Anim Res

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Objective: This study sought to determine the effectiveness of the cryodehydration technique in preserving the morphologic and morphometric attributes of the anatomical specimens of goats. Materials and methods: Different anatomical parts of a goat, i.e., heart, lungs, spleen, liver, kidney, and musculoskeletal specimens, were collected and fixed in 15% formalin. Later on, the fixed specimens were cryodehydrated by fast freezing (burning process) and repeated freezing-thawing sessions, followed by wood glue coating. Finally, the macroscopic (i.e., weight, color, texture, odor, and durability) and microscopic characteristics (by routine hematoxylin and eosin staining) of the cryodehydrated specimens were studied. Results: The resultant specimens produced excellent color and texture and were lightweight (60%–80% weight loss), soft, dry, odorless, durable, and easy to handle. The histoarchitectural details of the heart and skeletal muscle were well preserved, while some distinctive alterations were observed in the parenchymatous organs, i.e., breach in cellular integrity, loss of cell cytoplasm, loss of cytoplasmic and nuclear clarity, increased sinusoidal space, dilatation of the renal tubules, and reduction in glomerular size. Nevertheless, the basic histoarchitecture of each specimen was yet to be distinctly identifiable. Conclusion: The current study findings suggest that the cryodehydration technique can preserve gross anatomical features as well as histoarchitectural details and can be an effective tool for facilitating veterinary education and research.

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“…For example, the material must be beam stable, otherwise the contact with the ion and/or electron beams causes the structures of the sample to change, leading to inaccurate analysis. In life sciences, these samples are commonly chemically fixed and embedded in resin to impart resistance to the beam [3].…”

Section: Introductionmentioning

confidence: 99%

“…Fitting the FIB-SEM with cryo-technology allows the large volume (∼100µm 3 ) analysis of biological specimens suspended in vitreous ice [4,5]. This stabilises and perfectly preserves the structures of the specimen in their native state [3,6]. This also allows more accurate signal acquisition during experiment A scanning electron microscope is then used to obtain surface information from that newly revealed surface.…”

Section: Introductionmentioning

confidence: 99%

A Targeted Sampling Strategy for Compressive Cryo Focused Ion Beam Scanning Electron Microscopy

Nicholls¹,

Wells²,

Robinson³

et al. 2022

Preprint

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Cryo Focused Ion-Beam Scanning Electron Microscopy (cryo FIB-SEM) enables three-dimensional and nanoscale imaging of biological specimens via a slice and view mechanism. The FIB-SEM experiments are, however, limited by a slow (typically, several hours) acquisition process and the high electron doses imposed on the beam sensitive specimen can cause damage. In this work, we present a compressive sensing variant of cryo FIB-SEM capable of reducing the operational electron dose and increasing speed. We propose two Targeted Sampling (TS) strategies that leverage the reconstructed image of the previous sample layer as a prior for designing the next subsampling mask. Our image recovery is based on a blind Bayesian dictionary learning approach, i.e., Beta Process Factor Analysis (BPFA). This method is experimentally viable due to our ultra-fast GPU-based implementation of BPFA. Simulations on artificial compressive FIB-SEM measurements validate the success of proposed methods: the operational electron dose can be reduced by up to 20 times. These methods have large implications for the cryo FIB-SEM community, in which the imaging of beam sensitive biological materials without beam damage is crucial.

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A Brief Review of Cryobiology with Reference to Cryo Field Emission Scanning Electron Microscopy

Cited by 5 publications

References 159 publications

Fluorescence correlation spectroscopy and fluorescence lifetime imaging microscopy for deciphering the morphological evolution of supramolecular self-assembly

Fluorescence correlation spectroscopy and fluorescence lifetime imaging microscopy for deciphering the morphological evolution of supramolecular self-assembly

Efficacy of cryodehydration technique in preserving the gross and histoarchitectural details of goat visceral and musculoskeletal specimens

A Targeted Sampling Strategy for Compressive Cryo Focused Ion Beam Scanning Electron Microscopy

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