High accuracy, fiducial marker-based image registration of correlative microscopy images

Fluorescence microscopy (FM) and electron microscopy (EM) are complementary techniques. FM affords examination of large fields of view and identifying regions of interest but has a low resolution. EM exhibits excellent resolution over a limited field of view. The combination of these two techniques,...

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Bibliographic Details
Published in:Scientific reports 2019-03, Vol.9 (1), p.3211-3211, Article 3211
Main Authors: Mohammadian, Sajjad, Fokkema, Jantina, Agronskaia, Alexandra V., Liv, Nalan, de Heus, Cecilia, van Donselaar, Elly, Blab, Gerhard A., Klumperman, Judith, Gerritsen, Hans C.
Format: Article
Language:English
Subjects:
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101/47
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Accuracy
Electron microscopy
Fluorescence microscopy
Humanities and Social Sciences
Microscopes
Microscopy
multidisciplinary
Nanoparticles
Science
Science (multidisciplinary)
Silica
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Summary:Fluorescence microscopy (FM) and electron microscopy (EM) are complementary techniques. FM affords examination of large fields of view and identifying regions of interest but has a low resolution. EM exhibits excellent resolution over a limited field of view. The combination of these two techniques, correlative microscopy, received considerable interest in the past years and has proven its potential in biology and material science. Accurate correlation of FM and EM images is, however, challenging due to the differences in contrast mechanism, size of field of view and resolution. We report an accurate, fast and robust method to correlate FM and EM images using low densities of fiducial markers. Here, 120 nm diameter fiducial markers consisting of fluorescently labelled silica coated gold nanoparticles are used. The method relies on recording FM, low magnification EM and high magnification EM images. Two linear transformation matrices are constructed, FM to low magnification EM and low magnification EM to high magnification EM. Combination of these matrices results in a high accuracy transformation of FM to high magnification EM coordinates. The method was tested using two different transmission electron microscopes and different Tokuyasu and Lowicryl sections. The overall accuracy of the correlation method is high, 5–30 nm.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-019-40098-4