Simple Method of Specimen Preparation for Scanning Electron Microscopy

Buravkov, S. V.; Chernikov, Valery P.; Буравкова, Л. Б.

doi:10.1007/s10517-011-1335-7

Cited by 12 publications

(7 citation statements)

References 8 publications

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“…When samples are air-dried, surface tension occurring during evaporation of dehydrating fluids such as alcohol can cause substantial and irreversible deformation of structures (Buravkov et al, 2011). Therefore small and fragile life cycle stages such as microgamonts may not be identified because the air drying method could have caused them to collapse.…”

Section: Article In Pressmentioning

confidence: 99%

Validation of cell-free culture using scanning electron microscopy (SEM) and gene expression studies

Yang

Elankumaran

Hijjawi

et al. 2015

Experimental Parasitology

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• Scanning electron microscopy (SEM) characterisation of life cycle stages in cell-free culture.• Comparison of gene expression in cell culture and cell-free culture.• Gene expression cell-free culture was delayed and was lower.• Data provide support for cell-free culture of Cryptosporidium. A cell-free culture system for Cryptosporidium parvum was analysed using scanning electron microscopy (SEM) to characterise life cycle stages and compare gene expression in cell-free culture and cell culture using HCT-8 cells. Cryptosporidium parvum samples were harvested at 2 h, 8 h, 14 h, 26 h, 50 h, 74 h, 98 h, 122 h and 170 h, chemically fixed and specimens were observed using a Zeiss 1555 scanning electron microscope. The presence of sporozoites, trophozoites and type I merozoites were identified by SEM. Gene expression in cell culture and cell-free culture was studied using reverse transcriptase quantitative PCR (RT-qPCR) of the sporozoite surface antigen protein (cp15), the glycoprotein 900 (gp900), the Cryptosporidium oocyst wall protein (COWP) and 18S ribosomal RNA (rRNA) genes in both cell free and conventional cell culture. In cell culture, cp15 expression peaked at 74 h, gp900 expression peaked at 74 h and 98 h and COWP expression peaked at 50 h. In cell-free culture, CP15 expression peaked at 98 h, gp900 expression peaked at 74 h and COWP expression peaked at 122 h. The present study is the first to compare gene expression of C. parvum in cell culture and cell-free culture and to characterise life cycle stages of C. parvum in cell-free culture using SEM. Findings from this study showed that gene expression patterns in cell culture and cell-free culture were similar but in cell-free culture, gene expression was delayed for CP15 and COWP in cell free culture compared with the cell culture system and was lower. Although three life cycle stageswere conclusively identified, improvements in SEM methodology should lead to the detection of more life cycle stages. G R A P H I C A L A B S T R A C T

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Section: Article In Pressmentioning

confidence: 99%

Validation of cell-free culture using scanning electron microscopy (SEM) and gene expression studies

Yang

Elankumaran

Hijjawi

et al. 2015

Experimental Parasitology

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“…These two methods avoid producing the surface-tension and capillary effects which can adversely distort microscopic structure ( Favret 2009 ). Alternative chemical approaches include using solvents with low surface tension such as hexamethyldisilazane (HMDS) ( Nation 1983 , Slížová et al 2003 , Buravkov et al 2011 ) or Acetone ( Van Noort 1996 ). Both methods result in greatly reduced deformation of the soft structures in less sclerotized insects.…”

Section: Discussionmentioning

confidence: 99%

Preservation of iridescent colours in Phorinia Robineau-Desvoidy, 1830 (Diptera: Tachinidae)

Braet¹,

Downes²,

Simonis³

2016

BDJ

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BackgroundIridescent blue-green colours are exhibited by various organisms including several taxa in the Tachinidae (Diptera) with notable examples within the Afrotropical members of the genus Phorinia Robineau-Desvoidy, 1830. The vivid colouration observed in life quickly fades to a dull golden-yellow when a specimen is dried. Although well known, no published explanation has been given for this phenomenon.New informationWe illustrate the mechanism associated with this colour change. We also test and propose technical alternatives to retain the living colours in dried specimens.

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“…Recent studies have reported similar surface treatment approaches by skipping the metal sputtering steps in SEM imaging. [20][21][22][23][24] Several different methods, such as a plasma nitride layer, an osmium-thiocarbohydrazide-osmium layer, and ionic liquids, have been suggested as alternatives for sputter coating; however, ionic liquid and osmium-thiocarbohydrazide-osmium methods are more akin to shape preservation methods and thus require sample drying steps, and plasma nitrating can only be utilized for hard materials such as metals.…”

Section: Dehydration Prevention and Structural Supportmentioning

confidence: 99%

A nature-inspired protective coating on soft/wet biomaterials for SEM by aerobic oxidation of polyphenols

Park

Lee

et al. 2020

Mater. Horiz.

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Simple Method of Specimen Preparation for Scanning Electron Microscopy

Cited by 12 publications

References 8 publications

Validation of cell-free culture using scanning electron microscopy (SEM) and gene expression studies

Validation of cell-free culture using scanning electron microscopy (SEM) and gene expression studies

Preservation of iridescent colours in Phorinia Robineau-Desvoidy, 1830 (Diptera: Tachinidae)

A nature-inspired protective coating on soft/wet biomaterials for SEM by aerobic oxidation of polyphenols

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