Retrospective: Radiation damage and its associated “Information Limitations”

Glaeser, Robert M.

doi:10.1016/j.jsb.2008.06.001

Cited by 77 publications

(82 citation statements)

References 48 publications

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“…A new barrier emerged during the early 1970's with the realization that organic specimens could not be visualized directly in the electron microscope because either the electron beam destroyed the specimen or, if the image was recorded without damaging the specimen, it contained too few electrons to be interpreted directly (Glaeser, 1971;Glaeser, 2008). Negative stain had the useful property that, when exposed to high doses of electrons, specimens reached a relatively stable end point in which the protein would be obliterated but its outlines were still retained.…”

Section: Introductionmentioning

confidence: 99%

Retrospective on the early development of cryoelectron microscopy of macromolecules and a prospective on opportunities for the future

Taylor

Glaeser

2008

Journal of Structural Biology

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Methods for preserving specimen hydration in protein crystals were pursued in the early 1970s as a prerequisite for protein crystallography using an electron microscope. Three laboratories approached this question from very different directions. One built a differentially pumped hydration chamber that could maintain the crystal in a liquid water environment, a second maintained hydration by rapidly freezing the protein crystal and examining it in a cold stage, and the third replaced the water of hydration by using glucose in the same way as one had previously used "negative stains". Each of these early efforts succeeded in preserving the structures of protein crystals at high resolution within the vacuum of the electron microscope, as demonstrated by electron diffraction patterns. The next breakthrough came in the early 1980s when a technique was devised to preserve noncrystalline specimens by freezing them within vitreous ice. Since then, with the development of high stability cold stages and transfer mechanisms compatible with many instrument platforms, and by using commercially provided low-dose imaging techniques to avoid radiation damage, there has been an explosion of applications. These now include single particles, helical filaments, 2-D arrays and even whole cells, where the most exciting recent applications involve cryoelectron tomography. These achievements and possibilities generate a new set of research opportunities associated with increasing the reliability and throughput with which specimens can be studied by cryoEM.

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Section: Introductionmentioning

confidence: 99%

Retrospective on the early development of cryoelectron microscopy of macromolecules and a prospective on opportunities for the future

Taylor

Glaeser

2008

Journal of Structural Biology

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151

119

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“…The untoward effects of high-energy beam bombardment are exaggerated in the vacuum conditions typical to electron microscope systems (Stevens-Kalceff, 2004). Beam damage and beam-induced movement are fundamental restricts for resolution power of electron microscopy (Glaeser, 2008). Beam-induced heating is associated with thickening of the oxide layer on the material under study through electron beam-facilitated mass transport across the oxide layer within a defect-related circuit (Baker et al, 2010).…”

Section: Beam Damagementioning

confidence: 99%

“…The appropriate time and amount of electron exposure should be determined experimentally with the goal of sufficient exposures for obtaining useful signal-to-noise ratio and high-resolution images without image destruction (Baker et al, 2010). The relationship between inherent image contrast, applied exposure for image recording and image quality should be considered for getting a balanced state Glaeser, 2008). This is worth to note the fact that very high-quality images could nevertheless be obtained but averaging obtained images might be helpful in approximating the well-defined resolution limit (Glaeser, 2008).…”

Section: Beam Damagementioning

confidence: 99%

Six common errors cause dangerous mistakes in interpretation of electron micrographs

Moradi¹,

Behjati

2011

Microscopy Res & Technique

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The highly complex techniques of electron microscopy made it bound to the sensitive and critical micrograph analysis. The accurately interpreted micrographs are of paramount values in basic investigations. Interpretation skills and quality of the micrographs are the two fundamental keys in accomplishment of these goals but there are many mistakes and errors that can happen during the sample preparation, sectioning, EM operation, and photo publishing. The mentioned mistakes and errors effect directly in the final result which is a micrograph and can lead the analyzer who can be a pathologist to an interpretation followed by serious danger for patient. Artifacts caused by any given stimuli expected to be bothersome for investigators. Even best qualified equipments can be regarded as source of artifact generation. In this article, seven serious errors in electron micrographs which usually occur in transmission electron microscopy are addressed.

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“…Diese Schicht kann durch vorsichtige Bestrahlung mit einem Elektronenstrahl geringer Intensität nachgewiesen und entfernt werden (Abbildung 5 e-h); dabei ist darauf zu achten, dass die Integrität der darunter liegenden Probe nicht beschädigt wird. [24] Die schädigende Wirkung von Elektronenstrahlen kann durch Absenken der Probentemperatur verringert werden, beispielsweise um einen Faktor von 3-6 zwischen Raumtemperatur und der Temperatur flüssigen Stickstoffs. [25] Dennoch sind auch bei Kryo-TEM-Proben sowohl die eingebetteten Nanostrukturen als auch die glasartige Matrix (besonders bei organischen Lösungsmitteln) empfindlich gegen Elektronenstrahl-induzierte Veränderungen.…”

Section: Tem-abbildung Selbstorganisierter Strukturenunclassified

Abbildung selbstorganisierter Strukturen: Interpretation von TEM‐ und Kryo‐TEM‐Aufnahmen

2010

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In der Materialchemie werden in Lösung gezüchtete Nanostrukturen oft mithilfe der Transmissionselektronenmikroskopie (TEM) untersucht. Häufig werden die TEM‐Proben bei der Probenvorbereitung getrocknet und kontrastiert, und bei der Bildaufnahme selbst kommt es zur Wechselwirkung zwischen der Probe und dem Elektronenstrahl. Beide Prozesse rufen zur Vorsicht bei der Interpretation der erhaltenen elektronenmikroskopischen Aufnahmen auf. Alternativ dazu kann durch kryogene Probenpräparation und anschließende Abbildung mit kryogener Niederdosis‐TEM ein beinahe nativer solvatisierter Zustand konserviert werden. In unserem Kurzaufsatz geben wir eine kritische Analyse der Probenvorbereitung und, noch wichtiger, der Datenaufnahme und Interpretation von elektronenmikroskopischen Aufnahmen. Die Übersicht soll auch Leitlinien für die Anwendung von (Kryo‐)TEM als leistungsstarkes und zuverlässiges Hilfsmittel für die Analyse kolloidaler und selbstorganisierter Strukturen mit Abmessungen im Nanometerbereich bieten.

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Retrospective: Radiation damage and its associated “Information Limitations”

Cited by 77 publications

References 48 publications

Retrospective on the early development of cryoelectron microscopy of macromolecules and a prospective on opportunities for the future

Retrospective on the early development of cryoelectron microscopy of macromolecules and a prospective on opportunities for the future

Six common errors cause dangerous mistakes in interpretation of electron micrographs

Abbildung selbstorganisierter Strukturen: Interpretation von TEM‐ und Kryo‐TEM‐Aufnahmen

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