Immunolocalization of multiple membrane proteins on a carbon replica with STEM and EDX

A. Loukanov, N. Kamasawa, R. Danev, R. Shigemoto, K. Nagayama, Ultramicroscopy 110 (2010) 366–374.

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Abstract
We present a method for immunolabeling of multiple species of membrane proteins with high spatial resolution. It allows differentiation of equally sized very small markers with different chemical compositions, which leads to high labeling efficiency and reduces steric hindrance of closely spaced immunolabeled biomolecules. Markers such as CdSe/ZnS semiconductor quantum dots and colloidal gold particles are distinguished by differential contrast in high-angle annular detector dark-field STEM mode or by EDX microanalysis of their elemental contents. This method was tested by observation of labeled AMPA- and NMDA-type glutamate receptors on sodium-dodecyl-sulfate-digested replica prepared from rat hippocampus. To improve particle visibility and detectability, the replica films were made exclusively with carbon to avoid the high background of conventional platinum/carbon replica. Extension of the method is suggested by detection of 1.4 nm nanogold particles and its potential application in the biological imaging research.
Publishing Year
Date Published
2010-03-01
Journal Title
Ultramicroscopy
Volume
110
Issue
4
Page
366 - 374
IST-REx-ID

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Loukanov A, Kamasawa N, Danev R, Shigemoto R, Nagayama K. Immunolocalization of multiple membrane proteins on a carbon replica with STEM and EDX. Ultramicroscopy. 2010;110(4):366-374. doi:10.1016/j.ultramic.2010.01.016
Loukanov, A., Kamasawa, N., Danev, R., Shigemoto, R., & Nagayama, K. (2010). Immunolocalization of multiple membrane proteins on a carbon replica with STEM and EDX. Ultramicroscopy, 110(4), 366–374. https://doi.org/10.1016/j.ultramic.2010.01.016
Loukanov, Alexandre, Naomi Kamasawa, Radostin Danev, Ryuichi Shigemoto, and Kuniaki Nagayama. “Immunolocalization of Multiple Membrane Proteins on a Carbon Replica with STEM and EDX.” Ultramicroscopy 110, no. 4 (2010): 366–74. https://doi.org/10.1016/j.ultramic.2010.01.016.
A. Loukanov, N. Kamasawa, R. Danev, R. Shigemoto, and K. Nagayama, “Immunolocalization of multiple membrane proteins on a carbon replica with STEM and EDX,” Ultramicroscopy, vol. 110, no. 4, pp. 366–374, 2010.
Loukanov A, Kamasawa N, Danev R, Shigemoto R, Nagayama K. 2010. Immunolocalization of multiple membrane proteins on a carbon replica with STEM and EDX. Ultramicroscopy. 110(4), 366–374.
Loukanov, Alexandre, et al. “Immunolocalization of Multiple Membrane Proteins on a Carbon Replica with STEM and EDX.” Ultramicroscopy, vol. 110, no. 4, Elsevier, 2010, pp. 366–74, doi:10.1016/j.ultramic.2010.01.016.

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