Factors influencing the precision of quantitative scanning transmission electron microscopy

Müller, Shirley A.; Goldie, Kenneth N.; Bürki, Roland; Haring, Robert E.; Engel, Andréas

doi:10.1016/0304-3991(92)90022-c

Cited by 156 publications

(133 citation statements)

References 39 publications

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“…(b) Mass--per--length histogram of correspondingly written TMV. After scaling according to the mass standard, the determined average mass--per--length, 135 ± 6 kDa/nm, is within 1.5% of the expected value, 133 kDa/nm (Müller et al, 1992). Further, the standard deviation of the data set is comparable to that of the mass standard, ± 4 kDa/nm, which was prepared in the conventional way (see Materials and Methods).…”

Section: Discussionmentioning

confidence: 57%

“…All samples were prepared on thin carbon films spanning gold sputtered, carbon--coated, 200--mesh--per--inch, gold--plated copper grids (STEM microscopy grids) (Müller et al, 1992). The aim was to determine whether samples leaving the microfluidics setup were clean and suitable for mass evaluation.…”

Section: Scanning Transmission Electron Microscopymentioning

confidence: 99%

“…The MPL data from the test grid were binned into a histogram and described by a Gaussian curve. The average MPL and standard deviation (SD) were also calculated and compared to the expected value of 133 kDa/nm 2 (Müller et al, 1992), and to the SD of the control grid results, respectively.…”

Section: Scanning Transmission Electron Microscopymentioning

confidence: 99%

“…4b). Mass measurements require the removal of all non--volatile buffer components (Müller et al, 1992). The classic way to do this specific desalting is to wash the grid several times with quartz--ddH2O or a volatile buffer, e.g., ammonium acetate or bicarbonate, directly after sample adsorption.…”

Section: Fig 5)mentioning

confidence: 99%

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Connecting μ-fluidics to electron microscopy

Kemmerling

Ziegler

Schweighauser

et al. 2012

Journal of Structural Biology

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A versatile methodology for electron microscopy (EM) grid preparation enabling total content sample analysis is presented. A microfluidic-dialysis conditioning module to desalt or mix samples with negative stain solution is used, combined with a robotic writing table to micro-pattern the EM grids. The method allows heterogeneous samples of minute volumes to be processed at physiological pH for structure and mass analysis, and allows the preparation characteristics to be finely tuned.

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Section: Discussionmentioning

confidence: 57%

Section: Scanning Transmission Electron Microscopymentioning

confidence: 99%

Section: Scanning Transmission Electron Microscopymentioning

confidence: 99%

Section: Fig 5)mentioning

confidence: 99%

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Connecting μ-fluidics to electron microscopy

Kemmerling

Ziegler

Schweighauser

et al. 2012

Journal of Structural Biology

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“…However, the microtubule binding properties of dimeric motor constructs are more complicated to investigate than the ones from monomers. Recent cryo-EM and scanning transmission electron microscopy (STEM)-based mass determinations (Müller et al, 1992) demonstrated the complexity of dimeric kinesin interaction with microtubules Hoenger et al 2000). While there is agreement on the arrangement of dimeric ncd (a retrograde kinesin-like motor) on the microtubule surface (Sosa et al, 1997;Hirose et al, 1998), divergent interpretations have been given for the interaction of dimeric kinesin with microtubules (Arnal et al, 1996;Hirose et al, 1996Hirose et al, , 1999Hoenger et al, 1998Hoenger et al, , 2000.…”

Section: Introductionmentioning

confidence: 99%

Surface Topography of Microtubule Walls Decorated with Monomeric and Dimeric Kinesin Constructs

Hoenger

Doerhoefer²,

Woehlke³

et al. 2000

Biological Chemistry

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The surface topography of opened-up microtubule walls (sheets) decorated with monomeric and dimeric kinesin motor domains was investigated by freezedrying and unidirectional metal shadowing. Electron microscopy of surface-shadowed specimens produces images with a high signal/noise ratio, which enable a direct observation of surface features below 2 nm detail. Here we investigate the inner and outer surface of microtubules and tubulin sheets with and without decoration by kinesin motor domains. Tubulin sheets are flattened walls of microtubules, keeping lateral protofilament contacts intact. Surface shadowing reveals the following features: (i) when the microtubule outside is exposed the surface relief is dominated by the bound motor domains. Monomeric motor constructs generate a strong 8 nm periodicity, corresponding to the binding of one motor domain per ␣-␤-tubulin heterodimer. This surface periodicity largely disappears when dimeric kinesin motor domains are used for decoration, even though it is still visible in negatively stained or frozen hydrated specimens. This could be explained by disorder in the binding of the second (loosely tethered) kinesin head, and/or disorder in the coiled-coil tail. (ii) Both surfaces of undecorated sheets or microtubules, as well as the inner surface of decorated sheets, reveal a strong 4 nm repeat (due to the periodicity of tubulin monomers) and a weak 8 nm repeat (due to slight differences between ␣-and ␤-tubulin). The differences between ␣-and ␤-tubulin on the inner surface are stronger than expected from cryo-electron microscopy of unstained microtubules, indicating the existence of tubulin subdomain-specific surface properties that reflect the surface corrugation and hence metal deposition during evaporation. The 16 nm periodicity visible in some negatively stained specimens (caused by the pairing of cooperatively bound kinesin dimers) is not detected by surface shadowing.

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Microfibrillar elements of the dermal matrix

Kielty¹,

Shuttleworth²

1997

Microsc. Res. Tech.

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Connective tissue microfibrils are key structural elements of the dermal matrix which play major roles in establishing and maintaining the structural and mechanical integrity of this complex tissue. Type VI collagen microfibrils form extensive microfibrillar networks which intercalate between the major collagen fibrils and are juxtaposed to cellular basement membranes, blood vessels and other interstitial structures. Fibrillin microfibrils define the continuous elastic network of skin, and are present in dermis as microfibril bundles devoid of measureable elastin extending from the dermal‐epithelial junction and as components of the thick elastic fibres present in the deep reticular dermis. Electron microscopic analyses have revealed both classes of microfibrils to have complex ultrastructures. The ability to isolate intact native microfibrils from skin has enabled a combination of high resolution and biochemical techniques to be applied to elucidate their structure:function relationships. These approaches have generated new information about their molecular organisation and physiological interactions in health and disease. Microsc. Res. Tech. 38:413–427, 1997. © 1997 Wiley‐Liss, Inc.

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Factors influencing the precision of quantitative scanning transmission electron microscopy

Cited by 156 publications

References 39 publications

Connecting μ-fluidics to electron microscopy

Connecting μ-fluidics to electron microscopy

Surface Topography of Microtubule Walls Decorated with Monomeric and Dimeric Kinesin Constructs

Microfibrillar elements of the dermal matrix

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