Correlative Light and Electron Microscopy Using Frozen Section Obtained Using Cryo-Ultramicrotomy

Kim, Hong-Lim; Riew, Tae-Ryong; Park, Jieun; Lee, Youngchun; Kim, In-Beom

doi:10.3390/ijms22084273

Open AccessArticle

Correlative Light and Electron Microscopy Using Frozen Section Obtained Using Cryo-Ultramicrotomy

by

Hong-Lim Kim

¹,

Tae-Ryong Riew

²,

Jieun Park

¹,

Youngchun Lee

¹ and

In-Beom Kim

^1,2,3,4,*

¹

Integrative Research Support Center, College of Medicine, The Catholic University of Korea, Seoul 06591, Korea

²

Department of Anatomy, College of Medicine, The Catholic University of Korea, Seoul 06591, Korea

³

Catholic Neuroscience Institute, College of Medicine, The Catholic University of Korea, Seoul 06591, Korea

⁴

Catholic Institute for Applied Anatomy, College of Medicine, The Catholic University of Korea, Seoul 06591, Korea

^*

Author to whom correspondence should be addressed.

Int. J. Mol. Sci. 2021, 22(8), 4273; https://doi.org/10.3390/ijms22084273

Submission received: 19 March 2021 / Revised: 13 April 2021 / Accepted: 17 April 2021 / Published: 20 April 2021 / Corrected: 9 June 2022

(This article belongs to the Special Issue Applications of Advanced Imaging and Analytical Microscopic Techniques on Molecular and Cell Biology)

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Abstract

Immuno-electron microscopy (Immuno-EM) is a powerful tool for identifying molecular targets with ultrastructural details in biological specimens. However, technical barriers, such as the loss of ultrastructural integrity, the decrease in antigenicity, or artifacts in the handling process, hinder the widespread use of the technique by biomedical researchers. We developed a method to overcome such challenges by combining light and electron microscopy with immunolabeling based on Tokuyasu’s method. Using cryo-sectioned biological specimens, target proteins with excellent antigenicity were first immunolabeled for confocal analysis, and then the same tissue sections were further processed for electron microscopy, which provided a well-preserved ultrastructure comparable to that obtained using conventional electron microscopy. Moreover, this method does not require specifically designed correlative light and electron microscopy (CLEM) devices but rather employs conventional confocal and electron microscopes; therefore, it can be easily applied in many biomedical studies.

Keywords: correlative light and electron microscopy; immuno-electron microscopy; FluoroNanogold; cryo-ultramicrotomy

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MDPI and ACS Style

Kim, H.-L.; Riew, T.-R.; Park, J.; Lee, Y.; Kim, I.-B. Correlative Light and Electron Microscopy Using Frozen Section Obtained Using Cryo-Ultramicrotomy. Int. J. Mol. Sci. 2021, 22, 4273. https://doi.org/10.3390/ijms22084273

AMA Style

Kim H-L, Riew T-R, Park J, Lee Y, Kim I-B. Correlative Light and Electron Microscopy Using Frozen Section Obtained Using Cryo-Ultramicrotomy. International Journal of Molecular Sciences. 2021; 22(8):4273. https://doi.org/10.3390/ijms22084273

Chicago/Turabian Style

Kim, Hong-Lim, Tae-Ryong Riew, Jieun Park, Youngchun Lee, and In-Beom Kim. 2021. "Correlative Light and Electron Microscopy Using Frozen Section Obtained Using Cryo-Ultramicrotomy" International Journal of Molecular Sciences 22, no. 8: 4273. https://doi.org/10.3390/ijms22084273

APA Style

Kim, H.-L., Riew, T.-R., Park, J., Lee, Y., & Kim, I.-B. (2021). Correlative Light and Electron Microscopy Using Frozen Section Obtained Using Cryo-Ultramicrotomy. International Journal of Molecular Sciences, 22(8), 4273. https://doi.org/10.3390/ijms22084273

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

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Correlative Light and Electron Microscopy Using Frozen Section Obtained Using Cryo-Ultramicrotomy

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