Stop-Frame Filming and Discovery of Reactions at the Single-Molecule Level by Transmission Electron Microscopy

AbstractRecent advances in microscopy techniques enabled nanoscale discoveries in biology. In particular, electron microscopy reveals important cellular structures with nanometer resolution, yet it is hard, and sometimes impossible to resolve specific protein localizations. Super-resolution fluorescence microscopy techniques developed over the recent years allow for protein-specific localization with ~ 20 nm precision are overcoming this limitation, yet it remains challenging to place those in cells without a reference frame. Correlative light and electron microscopy (CLEM) approaches have been developed to place the fluorescence image in the context of a cellular structure. However, combining imaging methods such as super resolution microscopy and transmission electron microscopy necessitates a correlation using fiducial markers to locate the fluorescence on the structures visible in electron microscopy, with a measurable precision. Here, we investigated different fiducial markers for super-resolution CLEM (sCLEM) by evaluating their shape, intensity, stability and compatibility with photoactivatable fluorescent proteins as well as the electron density. We further carefully determined limitations of correlation accuracy. We found that spectrally-shifted FluoSpheres are well suited as fiducial markers for correlating single-molecule localization microscopy with transmission electron microscopy.

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Dual responsive specifically labelled carbogenic fluorescent nanodots for super resolution and electron microscopy

Nanoscale ◽

10.1039/c9nr00457b ◽

2019 ◽

Vol 11 (14) ◽

pp. 6561-6565 ◽

Cited By ~ 4

Author(s):

Navneet. C. Verma ◽

Chethana Rao ◽

Ashutosh Singh ◽

Neha Garg ◽

Chayan K. Nandi

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Single Molecule ◽

Super Resolution ◽

Dual Responsive ◽

Stochastic Optical Reconstruction Microscopy ◽

Transmission Electron ◽

Optical Reconstruction

We introduce an orange emissive fluorescent nanodot for successful single molecule stochastic optical reconstruction microscopy (STORM), super resolution radial fluctuation (SRRF) microscopy and transmission electron microscopy (TEM).

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Techniques for Transmission Electron Microscopy of Fiber-Matrix Interfaces in Aluminum Alloy-Boron Composites by Ion Erosio

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100065079 ◽

1971 ◽

Vol 29 ◽

pp. 204-205

Author(s):

G. G. Shaw

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Aluminum Alloy ◽

Electron Beam ◽

Pure Aluminum ◽

Ion Milling ◽

Transmission Electron ◽

Fiber Matrix ◽

Ion Erosion ◽

Row Of Fibers

The morphology and composition of the fiber-matrix interface can best be studied by transmission electron microscopy and electron diffraction. For some composites satisfactory samples can be prepared by electropolishing. For others such as aluminum alloy-boron composites ion erosion is necessary.When one wishes to examine a specimen with the electron beam perpendicular to the fiber, preparation is as follows: A 1/8 in. disk is cut from the sample with a cylindrical tool by spark machining. Thin slices, 5 mils thick, containing one row of fibers, are then, spark-machined from the disk. After spark machining, the slice is carefully polished with diamond paste until the row of fibers is exposed on each side, as shown in Figure 1.In the case where examination is desired with the electron beam parallel to the fiber, preparation is as follows: Experimental composites are usually 50 mils or less in thickness so an auxiliary holder is necessary during ion milling and for easy transfer to the electron microscope. This holder is pure aluminum sheet, 3 mils thick.

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Direct Observation of Heat Exchanger Deposits by Cross Sectioning

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100061835 ◽

1967 ◽

Vol 25 ◽

pp. 364-365

Author(s):

R. W. Anderson ◽

D. L. Senecal

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Heat Exchanger ◽

Direct Observation ◽

Cross Sections ◽

Preparation Method ◽

Specimen Preparation ◽

Flowing Water ◽

Transmission Electron ◽

Deposit Layer

A problem was presented to observe the packing densities of deposits of sub-micron corrosion product particles. The deposits were 5-100 mils thick and had formed on the inside surfaces of 3/8 inch diameter Zircaloy-2 heat exchanger tubes. The particles were iron oxides deposited from flowing water and consequently were only weakly bonded. Particular care was required during handling to preserve the original formations of the deposits. The specimen preparation method described below allowed direct observation of cross sections of the deposit layers by transmission electron microscopy.The specimens were short sections of the tubes (about 3 inches long) that were carefully cut from the systems. The insides of the tube sections were first coated with a thin layer of a fluid epoxy resin by dipping. This coating served to impregnate the deposit layer as well as to protect the layer if subsequent handling were required.

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Phase Transformations in Ti-7w/oMo-3w/oMe Alloy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100052584 ◽

1974 ◽

Vol 32 ◽

pp. 514-515

Author(s):

S. Fujishiro

Keyword(s):

Electron Microscopy ◽

Mechanical Properties ◽

Transmission Electron Microscopy ◽

Tensile Strength ◽

Mechanical Processing ◽

Ray Method ◽

X Ray ◽

Transmission Electron ◽

Water Quench ◽

Alpha Beta

The mechanical properties of three titanium alloys (Ti-7Mo-3Al, Ti-7Mo- 3Cu and Ti-7Mo-3Ta) were evaluated as function of: 1) Solutionizing in the beta field and aging, 2) Thermal Mechanical Processing in the beta field and aging, 3) Solutionizing in the alpha + beta field and aging. The samples were isothermally aged in the temperature range 300° to 700*C for 4 to 24 hours, followed by a water quench. Transmission electron microscopy and X-ray method were used to identify the phase formed. All three alloys solutionized at 1050°C (beta field) transformed to martensitic alpha (alpha prime) upon being water quenched. Despite this heavily strained alpha prime, which is characterized by microtwins the tensile strength of the as-quenched alloys is relatively low and the elongation is as high as 30%.

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Two- or three-way observations of the identical cell elements within the same sections by LM, TEM and/or SEM

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100081279 ◽

1978 ◽

Vol 36 (2) ◽

pp. 82-83 ◽

Cited By ~ 1

Author(s):

Nakazo Watari ◽

Yasuaki Hotta ◽

Yoshio Mabuchi

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Transmission Electron Microscopy ◽

Fine Structure ◽

Light Microscopy ◽

Thin Sections ◽

Transmission Electron ◽

Identical Cell ◽

Electron Microscopes ◽

Scanning Electron

It is very useful if we can observe the identical cell elements within the same sections by light microscopy (LM), transmission electron microscopy (TEM) and/or scanning electron microscopy (SEM) sequentially, because, the cell fine structure can not be indicated by LM, while the color is; on the other hand, the cell fine structure can be very easily observed by EM, although its color properties may not. However, there is one problem in that LM requires thick sections of over 1 μm, while EM needs very thin sections of under 100 nm. Recently, we have developed a new method to observe the same cell elements within the same plastic sections using both light and transmission (conventional or high-voltage) electron microscopes.In this paper, we have developed two new observation methods for the identical cell elements within the same sections, both plastic-embedded and paraffin-embedded, using light microscopy, transmission electron microscopy and/or scanning electron microscopy (Fig. 1).

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Comparative Microstructures of Ion Milled and Electrochemically Thinned Composite Materials

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100052742 ◽

1974 ◽

Vol 32 ◽

pp. 546-547

Author(s):

R.R. Russell

Keyword(s):

Electron Microscopy ◽

Transmission Electron Microscopy ◽

Composite Materials ◽

Great Difficulty ◽

Ion Milling ◽

Transmission Electron ◽

Ion Damage ◽

Intermetallic Composite

Transmission electron microscopy of metallic/intermetallic composite materials is most challenging since the microscopist typically has great difficulty preparing specimens with uniform electron thin areas in adjacent phases. The application of ion milling for thinning foils from such materials has been quite effective. Although composite specimens prepared by ion milling have yielded much microstructural information, this technique has some inherent drawbacks such as the possible generation of ion damage near sample surfaces.

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