TY - JOUR
T1 - Correction to
T2 - Evaluation of serum extracellular vesicle isolation methods for profiling miRNAs by next-generation sequencing (Journal of Extracellular Vesicles, (2018), 7, 1, (1481321), 10.1080/20013078.2018.1481321)
AU - Buschmann, Dominik
AU - Kirchner, Benedikt
AU - Hermann, Stefanie
AU - Märte, Melanie
AU - Wurmser, Christine
AU - Brandes, Florian
AU - Kotschote, Stefan
AU - Bonin, Michael
AU - Steinlein, Ortrud K.
AU - Pfaffl, Michael W.
AU - Schelling, Gustav
AU - Reithmair, Marlene
N1 - Publisher Copyright:
© 2019, © 2019 Informa UK Limited, trading as Taylor & Francis Group.
PY - 2019/1/1
Y1 - 2019/1/1
N2 - Following the publication of this paper on 04 Jun 2018, it has come to the authors’ attention that the images (Figure 7) acquired in transmission electron microscopy (TEM) might not have convincingly displayed extracellular vesicles (EVs). During consultations with editors of the Journal of Extracellular Vesicles as well as experts in the field, it became clear that the initial protocol for TEM sample preparation lacked a methyl cellulose embedding step that is essential to maintain the structural integrity of EVs [1]. (Figure presented.) The authors have added this step for the improved protocol detailed below and herein present a revised Figure 7, which now convincingly depicts EVs. The morphology of EVs from each isolation method can be observed more clearly, and differences in their size, shape and levels of background contamination are more apparent. The structures observed in exoRNeasy isolates look distinctly different from those isolated by all other methods and reflect the increased particle diameters detected in nanoparticle tracking analysis (Figure 6). The published online version has been corrected to display a revised Figure 7. A correction has also been made to the Materials and methods section, paragraph Transmission electron microscopy: EVs were adsorbed onto formvar/carbon-coated 200-mesh nickel grids (Electron Microscopy Sciences) for 20 min at RT. Next, grids were fixed with 2% paraformaldehyde for 20 min, washed with PBS three times and fixed with 1 % glutaraldehyde for 5 min. After six washing steps with distilled water, grids were negative stained with 4 % uranyl acetate for 5 min. Finally, grids were embedded in 0.4 % uranyl acetate/0.2 % methyl cellulose for 10 min on ice in the dark and air-dried overnight. Images were acquired on a Zeiss EM900 (Carl Zeiss Microscopy GmbH) with a wide-angle dual-speed 2K-CCD camera at 80 kV. The authors apologize for any inconvenience caused by this discrepancy.
AB - Following the publication of this paper on 04 Jun 2018, it has come to the authors’ attention that the images (Figure 7) acquired in transmission electron microscopy (TEM) might not have convincingly displayed extracellular vesicles (EVs). During consultations with editors of the Journal of Extracellular Vesicles as well as experts in the field, it became clear that the initial protocol for TEM sample preparation lacked a methyl cellulose embedding step that is essential to maintain the structural integrity of EVs [1]. (Figure presented.) The authors have added this step for the improved protocol detailed below and herein present a revised Figure 7, which now convincingly depicts EVs. The morphology of EVs from each isolation method can be observed more clearly, and differences in their size, shape and levels of background contamination are more apparent. The structures observed in exoRNeasy isolates look distinctly different from those isolated by all other methods and reflect the increased particle diameters detected in nanoparticle tracking analysis (Figure 6). The published online version has been corrected to display a revised Figure 7. A correction has also been made to the Materials and methods section, paragraph Transmission electron microscopy: EVs were adsorbed onto formvar/carbon-coated 200-mesh nickel grids (Electron Microscopy Sciences) for 20 min at RT. Next, grids were fixed with 2% paraformaldehyde for 20 min, washed with PBS three times and fixed with 1 % glutaraldehyde for 5 min. After six washing steps with distilled water, grids were negative stained with 4 % uranyl acetate for 5 min. Finally, grids were embedded in 0.4 % uranyl acetate/0.2 % methyl cellulose for 10 min on ice in the dark and air-dried overnight. Images were acquired on a Zeiss EM900 (Carl Zeiss Microscopy GmbH) with a wide-angle dual-speed 2K-CCD camera at 80 kV. The authors apologize for any inconvenience caused by this discrepancy.
UR - http://www.scopus.com/inward/record.url?scp=85062636363&partnerID=8YFLogxK
U2 - 10.1080/20013078.2019.1581487
DO - 10.1080/20013078.2019.1581487
M3 - Comment/debate
AN - SCOPUS:85062636363
SN - 2001-3078
VL - 8
JO - Journal of Extracellular Vesicles
JF - Journal of Extracellular Vesicles
IS - 1
M1 - 1581487
ER -