Sample Preparation for Scanning Electron Microscopy: The Surprising Case of Freeze Drying from Tertiary Butanol

Baskin, Tobias I.; Orr, Teri J.; Jercinovic, Michael J.; Yoshida, Masato

doi:10.1017/s1551929514000522

Cited by 12 publications

(14 citation statements)

References 10 publications

(13 reference statements)

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“…Our studies support the idea that freeze-drying with the t-butanol method is a valuable alternative method to the CPD methods. This observation was consistent with previous studies which suggested that t-butanol is equivalent or sometimes superior to CPD (Inoué and Osatake, 1988;Baskin et al, 2014). However, when using vacuum freeze-drying from t-butanol, the pump oil needs to be changed more frequently since t-butanol damages the pump oil.…”

Section: Discussionsupporting

confidence: 92%

“…The t-butanol freeze-drying method (Inoué and Osatake, 1988) is a conventional method that is considered to yield good samples. It was reported that t-butanol is equivalent or sometimes superior to CPD (Inoué and Osatake, 1988;Baskin et al, 2014). The t-butanol freeze-drying method is commonly used to prepare biological SEM samples in Japan (Kaneko et al, 1990;Tuji, 2000;Watanabe et al, 2015), China (Xin et al, 2012), and other regions (El Sharaby et al, 2012).…”

Section: Conventional Sem T-butanol Freeze-drying Methodsmentioning

confidence: 99%

“…Methanol is generally considered to be the first choice as an SEM fixative. The t-butanol freeze-drying method of Baskin et al (2014) also used methanol as the fixative. There remains a lack of studies on the effect of different fixatives on the leaf morphology and dimension when a final freeze-drying with t-butanol is used for SEM preparation.…”

Section: Conventional Sem T-butanol Freeze-drying Methodsmentioning

confidence: 99%

“…This metal-coating step is necessary to avoid the accumulation of electron charges on sample surfaces, which are non-conductive. Numerous studies have been reported over the years and methods have been modified to become simpler, faster, and produce better results (Neinhuis and Edelmann, 1996;Talbot and White, 2013a;Talbot and White, 2013b;Baskin et al, 2014). Although much progress has been made, the conventional sample preparation procedure is complicated and time consuming, and might induce the shrinkage and collapse of epidermis cells.…”

Section: Introductionmentioning

confidence: 99%

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Comparison of Sample Preparation Techniques for Inspection of Leaf Epidermises Using Light Microscopy and Scanning Electronic Microscopy

et al. 2020

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Section: Discussionsupporting

confidence: 92%

Section: Conventional Sem T-butanol Freeze-drying Methodsmentioning

confidence: 99%

Section: Conventional Sem T-butanol Freeze-drying Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Comparison of Sample Preparation Techniques for Inspection of Leaf Epidermises Using Light Microscopy and Scanning Electronic Microscopy

et al. 2020

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“…The next day, the fixed samples were warmed to room temperature and ethanol dehydration was continued. The samples were dried using the tertiary butanol method 56 . After drying, the specimens were mounted on aluminum stubs and sputter-coated with gold using a Polaron E5100 sputter coater.…”

Section: Methodsmentioning

confidence: 99%

The importance of filamentous cyanobacteria in the development of oxygenic photogranules

Milferstedt

Kuo-Dahab

Butler

et al. 2017

Sci Rep

183

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Microorganisms often respond to their environment by growing as densely packed communities in biofilms, flocs or granules. One major advantage of life in these aggregates is the retention of its community in an ecosystem despite flowing water. We describe here a novel type of granule dominated by filamentous and motile cyanobacteria of the order Oscillatoriales. These bacteria form a mat-like photoactive outer layer around an otherwise unconsolidated core. The spatial organization of the phototrophic layer resembles microbial mats growing on sediments but is spherical. We describe the production of these oxygenic photogranules under static batch conditions, as well as in turbulently mixed bioreactors. Photogranulation defies typically postulated requirements for granulation in biotechnology, i.e., the need for hydrodynamic shear and selective washout. Photogranulation as described here is a robust phenomenon with respect to inoculum characteristics and environmental parameters like carbon sources. A bioprocess using oxygenic photogranules is an attractive candidate for energy-positive wastewater treatment as it biologically couples CO2 and O2 fluxes. As a result, the external supply of oxygen may become obsolete and otherwise released CO2 is fixed by photosynthesis for the production of an organic-rich biofeedstock as a renewable energy source.

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Melanin Treatment Effect of Vacuoles-Zinc Oxide Nanoparticles Combined with Ascorbic Acid

et al. 2022

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Currently, ascorbic acid (AA) is widely used as a skin whitening material, but, AA, an unstable hydrophilic molecule, cannot penetrate the skin easily, due to the hydrophobic character of the stratum corneum. Therefore, we conjugated AA with hydrated zinc oxide—an inorganic matrix with positive surface charge, to improve the stability of AA. The metal-conjugated-ascorbic acid (ZnAA) was then combined with yeast vacuole through the vacuolar membrane proteins that relate to metal transportation to create an enhanced vacuole that contained ZnAA. The characteristics of vacuole with ZnAA (ZnAA_Vac) were next examined by various tests that included X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT–IR), Field emission scanning electron microscopy (FE–SEM), and energy-dispersive X-ray (EDX) analysis. Furthermore, the ability of ZnAA_Vac to degrade melanin was confirmed in both melanoma cell line B16F10, and the artificial human skin MelanoDerm. The results showed that ZnAA_Vac possessed a higher depigmenting effect than the wild-type vacuole or ascorbic acid by reducing 75% of melanin color. Interestingly, ZnAA_Vac was found to be harmless, and did not cause any cytotoxicity to the cells. Overall, ZnAA_Vac is expected to provide a robust, harmless, and effective whitening agent for the skin.

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Sample Preparation for Scanning Electron Microscopy: The Surprising Case of Freeze Drying from Tertiary Butanol

Cited by 12 publications

References 10 publications

Comparison of Sample Preparation Techniques for Inspection of Leaf Epidermises Using Light Microscopy and Scanning Electronic Microscopy

Comparison of Sample Preparation Techniques for Inspection of Leaf Epidermises Using Light Microscopy and Scanning Electronic Microscopy

The importance of filamentous cyanobacteria in the development of oxygenic photogranules

Melanin Treatment Effect of Vacuoles-Zinc Oxide Nanoparticles Combined with Ascorbic Acid

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