Six common errors cause dangerous mistakes in interpretation of electron micrographs

Moradi, Iman; Behjati, Mohaddeseh

doi:10.1002/jemt.21111

Cited by 2 publications

(1 citation statement)

References 19 publications

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“…Thus, fixation protocols, permeabilization, contrast agents and fluorescent labelling approaches and should be carefully chosen and optimized for the specific research objective so as to obtain clean, high-resolution images for processing. Sample preparation methods and imaging techniques are discussed in depth for scanning electron (Goldberg, 2008;Moradi and Behjati, 2012), atomic force (El Kirat et al, 2005), and fluorescent microscopy (Hanrahan et al, 2011).…”

Section: Sample Processingmentioning

confidence: 99%

An insight into morphometric descriptors of cell shape that pertain to regenerative medicine

Lobo

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Biggs

et al. 2015

J Tissue Eng Regen Med

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Cellular morphology has recently been indicated as a powerful indicator of cellular function. The analysis of cell shape has evolved from rudimentary forms of microscopic visual inspection to more advanced methodologies that utilize high-resolution microscopy coupled with sophisticated computer hardware and software for data analysis. Despite this progress, there is still a lack of standardization in quantification of morphometric parameters. In addition, uncertainty remains as to which methodologies and parameters of cell morphology will yield meaningful data, which methods should be utilized to categorize cell shape, and the extent of reliability of measurements and the interpretation of the resulting analysis. A large range of descriptors has been employed to objectively assess the cellular morphology in two-dimensional and three-dimensional domains. Intuitively, simple and applicable morphometric descriptors are preferable and standardized protocols for cell shape analysis can be achieved with the help of computerized tools. In this review, cellular morphology is discussed as a descriptor of cellular function and the current morphometric parameters that are used quantitatively in two- and three-dimensional environments are described. Furthermore, the current problems associated with these morphometric measurements are addressed. Copyright © 2015 John Wiley & Sons, Ltd.

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Section: Sample Processingmentioning

confidence: 99%

An insight into morphometric descriptors of cell shape that pertain to regenerative medicine

Lobo

See

Biggs

et al. 2015

J Tissue Eng Regen Med

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Semi-quantitative monitoring of confluence of adherent mesenchymal stromal cells on calcium-phosphate granules by using widefield microscopy images

Piccinini

Pierini

Lucarelli

et al. 2014

J Mater Sci: Mater Med

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The analysis of cell confluence and proliferation is essential to design biomaterials and scaffolds to use as bone substitutes in clinical applications. Accordingly, several approaches have been proposed in the literature to estimate the area of the scaffold covered by cells. Nevertheless, most of the approaches rely on sophisticated equipment not employed for routine analyses, while the rest of them usually do not provide significant statistics about the cell distribution. This research aims at studying confluence and proliferation of mesenchymal stromal cells (MSC) adherent on OSPROLIFE(®), a commercial biomaterial in the form of granules. In particular, we propose a Computer Vision approach that can routinely be employed to monitor the surface of the single granules covered by cells because only a standard widefield fluorescent microscope is required. In order to acquire significant statistics data, we analyse wide-area images built by using MicroMos v2.0, an updated version of a previously published software specific for stitching brightfield and phase-contrast images manually acquired via a widefield microscope. In particular, MicroMos v2.0 permits to build accurate "mosaics" of fluorescent images, after correcting vignetting and photo-bleaching effects, providing a consistent representation of a sample region containing numerous granules. Then, our method allows to make automatically a statistically significant estimate of the percentage of the area of the single granules covered by cells. Finally, by analysing hundreds of granules at different time intervals we also obtained reliable data regarding cell proliferation, confirming that not only MSC adhere onto the OSPROLIFE(®) granules, but even proliferate over time.

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Six common errors cause dangerous mistakes in interpretation of electron micrographs

Cited by 2 publications

References 19 publications

An insight into morphometric descriptors of cell shape that pertain to regenerative medicine

An insight into morphometric descriptors of cell shape that pertain to regenerative medicine

Semi-quantitative monitoring of confluence of adherent mesenchymal stromal cells on calcium-phosphate granules by using widefield microscopy images

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