A specimen holder for low‐temperature scanning electron microscopy

Bastacky, J.; Goodman, Christian; Hayes, T.

doi:10.1002/jemt.1060140113

Cited by 6 publications

(2 citation statements)

References 2 publications

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“…Using a mechanical device to mount a frozen specimen also has difficulties. In most cases (e.g., in Bastacky et al 1990), the frozen sample is merely attached by clamping into a vice-like device using screws. Unlike adhesives, this procedure is not temperature dependent.…”

Section: Sample Preparationmentioning

confidence: 99%

Cryo‐planing for cryo‐scanning electron microscopy

Nijsse

Aelst

1999

Scanning

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Summary:In the past decade, investigators of cryo-planing for low-temperature scanning electron microscopy (cryo-SEM) have developed techniques that enable observations of flat sample surfaces. This study reviews these sample preparation techniques, compares and contrasts their results, and introduces modifications that improve results from cryo-planing. A prerequisite for all successful cryo-planing required a stable attachment of the specimen to a holder. In most cases, clamping with a screw mechanism and using indium as space-filler sufficed. Once this problem was solved, any of three existing cryo-planing methods could be used to provide successful results: cryo-milling, microtomy in a cold room, and cryo-ultramicrotomy. This study introduces modifications to the cryo-planing technique that produces flat surfaces of any desired plane through a specimen. These flat surfaces of frozen, fully hydrated samples can be used to improve observations from cryo-SEM as well as to enhance results from x-ray microanalysis and (digital) image analysis. Cryo-planing results of chrysanthemum (Dendranthema x grandiflorum Tzvelev) stems, hazel (Corylus avelane L.) stems, and rapeseed (Brassica napus L.) pistils are presented to illustrate the use of the planing method on fibrous, hard, and delicate materials, respectively.

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Section: Sample Preparationmentioning

confidence: 99%

Cryo‐planing for cryo‐scanning electron microscopy

Nijsse

Aelst

1999

Scanning

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“…Small sample holders with a low thermal mass are required to achieve very rapid specimen freezing rates (Wergin & Erbe, 1989b;Walther et al, 1990). It is also possible to mount specimens on a stub after cryofixation either by mechanical means (Bastacky et al, 1987a(Bastacky et al, , 1990 or by using a cryoadhesive with a suitable low-temperature freezing point (e.g. silver paint or carbon cement ;Ryan et al, 1988).…”

Section: P R E P a R A T I O N P R O T O C O L Smentioning

confidence: 99%

Low‐temperature scanning electron microscopy in biology

Read

Jeffree

1991

Journal of Microscopy

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SUMMARY A review of low‐temperature scanning electron microscopy (LTSEM) with regard to preparation protocols, specimen preservation, experimental approaches, and high‐resolution studies, is provided. Preparative procedures are described and recent developments in methodologies highlighted. It is now well established that LTSEM, for most biological specimens, provides superior specimen preservation than does ambient‐temperature SEM. This is because frozen‐hydrated samples retain most or all of their water, are rapidly immobilized and stabilized by cryofixation, and are not exposed to chemical modification or solvent extraction. Nevertheless, artefacts in LTSEM are common and most arise because frozen‐hydrated specimens contain water. LTSEM can be used as a powerful experimental tool. Advantages of employing LTSEM for this purpose and ways in which it can be used for novel experimentation are discussed. The most exciting development in recent years has been high‐resolution LTSEM. The advantages, problems and requirements for this approach are defined.

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