Integrated fluorescence and transmission electron microscopy

Agronskaia, Alexandra V.; Valentijn, Jack A.; Driel, Linda F. van; Schneijdenberg, Chris T.W.M.; Humbel, Bruno M.; Henegouwen, Paul M.P. van Bergen en; Verkleij, Arie J.; Koster, Abraham J.; Gerritsen, Hans C.

doi:10.1016/j.jsb.2008.07.003

Cited by 159 publications

(125 citation statements)

References 27 publications

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“…CLEM combines the advantages of these two techniques to allow direct examination of fluorescence localization within the high-resolution landscape of EM images. Recent advances in probe development, sample preparation and instrumentation are making CLEM more widely accessible (Darcy et al, 2006;Agronskaia et al, 2008;Cortese et al, 2009;Plitzko et al, 2009;Briegel et al, 2010;Bos et al, 2011;Bushby et al, 2011).…”

Section: Correlative Light and Electron Microscopymentioning

confidence: 99%

Advances in high-resolution imaging – techniques for three-dimensional imaging of cellular structures

Lidke

2012

Journal of Cell Science

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SummaryA fundamental goal in biology is to determine how cellular organization is coupled to function. To achieve this goal, a better understanding of organelle composition and structure is needed. Although visualization of cellular organelles using fluorescence or electron microscopy (EM) has become a common tool for the cell biologist, recent advances are providing a clearer picture of the cell than ever before. In particular, advanced light-microscopy techniques are achieving resolutions below the diffraction limit and EM tomography provides highresolution three-dimensional (3D) images of cellular structures. The ability to perform both fluorescence and electron microscopy on the same sample (correlative light and electron microscopy, CLEM) makes it possible to identify where a fluorescently labeled protein is located with respect to organelle structures visualized by EM. Here, we review the current state of the art in 3D biological imaging techniques with a focus on recent advances in electron microscopy and fluorescence super-resolution techniques.

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Section: Correlative Light and Electron Microscopymentioning

confidence: 99%

Advances in high-resolution imaging – techniques for three-dimensional imaging of cellular structures

Lidke

2012

Journal of Cell Science

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“…Commercially available solutions are already available for integrated X-ray microtomography (micro-CT) and scanning electron microscopy. Then there is the elegant paper by Agronkaia and colleagues who demonstrated the first fully integrated imaging with fluorescence microscopy and TEM in a single column (Agronskaia et al 2008) and the recent report on a fully integrated light optical-and scanning electron microscopy platform (Nishiyama et al 2010). This does not mean, however, that all the efforts in the development of CM tools were a waste of time.…”

Section: Conclusion and Future Outlooksmentioning

confidence: 99%

Multi-dimensional correlative imaging of subcellular events: combining the strengths of light and electron microscopy

Nykanen

Jahn

et al. 2010

Biophys Rev

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To genuinely understand how complex biological structures function, we must integrate knowledge of their dynamic behavior and of their molecular machinery. The combined use of light or laser microscopy and electron microscopy has become increasingly important to our understanding of the structure and function of cells and tissues at the molecular level. Such a combination of two or more different microscopy techniques, preferably with different spatial-and temporal-resolution limits, is often referred to as 'correlative microscopy'. Correlative imaging allows researchers to gain additional novel structurefunction information, and such information provides a greater degree of confidence about the structures of interest because observations from one method can be compared to those from the other method(s). This is the strength of correlative (or 'combined') microscopy, especially when it is combined with combinatorial or non-combinatorial labeling approaches. In this topical review, we provide a brief historical perspective of correlative microscopy and an in-depth overview of correlative sample-preparation and imaging methods presently available, including future perspectives on the trend towards integrative microscopy and microanalysis.

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“…Tannic acid [69] and uranyl acetate [44,87] may be incorporated as secondary or tertiary fixatives to improve membrane contrast. However, infiltration of chemicals can be slow and limits sample size to approximately 1 mm 3 . Microwave-accelerated immobilisation [100] has been used to increase the penetration rate of chemicals into samples and improve preservation through volumes.…”

Section: Overviewmentioning

confidence: 99%

“…Direct imaging of fluorescent molecules in situ in an EM is the ultimate realisation of CLEM. Integrated laser electron microscopy has taken the first steps towards this by feeding a laser line into the goniometer of a TEM [3] to image UVinduced nuclear structures in apoptotic cells [53].…”

Section: Locating Areas Of Interestmentioning

confidence: 99%

Towards native-state imaging in biological context in the electron microscope

2009

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Modern cell biology is reliant on light and fluorescence microscopy for analysis of cells, tissues and protein localisation. However, these powerful techniques are ultimately limited in resolution by the wavelength of light. Electron microscopes offer much greater resolution due to the shorter effective wavelength of electrons, allowing direct imaging of sub-cellular architecture. The harsh environment of the electron microscope chamber and the properties of the electron beam have led to complex chemical and mechanical preparation techniques, which distance biological samples from their native state and complicate data interpretation. Here we describe recent advances in sample preparation and instrumentation, which push the boundaries of high-resolution imaging. Cryopreparation, cryoelectron microscopy and environmental scanning electron microscopy strive to image samples in near native state. Advances in correlative microscopy and markers enable high-resolution localisation of proteins. Innovation in microscope design has pushed the boundaries of resolution to atomic scale, whilst automatic acquisition of high-resolution electron microscopy data through large volumes is finally able to place ultrastructure in biological context.

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Integrated fluorescence and transmission electron microscopy

Cited by 159 publications

References 27 publications

Advances in high-resolution imaging – techniques for three-dimensional imaging of cellular structures

Advances in high-resolution imaging – techniques for three-dimensional imaging of cellular structures

Multi-dimensional correlative imaging of subcellular events: combining the strengths of light and electron microscopy

Towards native-state imaging in biological context in the electron microscope

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