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DNA Origami Barcodes for Immunostaining
Praneetha Sundar Prakash1, Foram M Joshi2, Eric Vogelsberg2
1Department of Physics, 103 Smith Hall, Kent State University, Kent, Ohio 44242, United States.
Insights
This study introduces DNA origami barcodes for multiplexed immuno-gold labeling in electron microscopy. This novel approach enhances the ability to visualize multiple antigens simultaneously in biological samples.
Area of Science:
- Histology and Nanotechnology
- Biomedical Imaging
- Molecular Biology
Background:
- Immunostaining is crucial for studying cellular processes in histology.
- Immuno-gold labeling offers nanometer spatial resolution but struggles with multiplexing.
- Simultaneous visualization of multiple antigens remains a significant challenge in electron microscopy.
Purpose of the Study:
- To develop a DNA nanotechnology-based method for multiplexed antigen labeling in transmission electron microscopy.
- To create high-contrast patterns for visualizing multiple antigens simultaneously.
- To enhance the multiplexing capabilities of immunostaining in a modular fashion.
Main Methods:
- Utilized DNA origami structures as visual barcodes functionalized with gold nanoparticles.
- Developed DNA-modified antibodies designed to hybridize with complementary DNA strands on the barcodes.
- Applied the technique to ultrathin tissue resin sections for transmission electron microscopy imaging.
Main Results:
- Demonstrated a novel DNA nanotechnology approach for high-contrast antigen labeling.
- Successfully visualized multiple antigens in parallel using DNA origami barcodes.
- Showcased the modularity and expanded multiplexing potential of the developed technique.
Conclusions:
- The DNA nanotechnology-based approach offers a powerful solution for multiplexed immuno-gold labeling.
- This method significantly advances the ability to study complex cellular processes by enabling simultaneous visualization of multiple antigens.
- The modular design promises broad applicability in various fields of biological research requiring high-resolution imaging.
Abstract:
In histology, immunostaining of biological samples is a gold standard for studying cellular processes, such as the expression of cell surface markers or the cellular uptake of proteins and drug molecules. Immuno-gold labeling is a commonly used technique to achieve nanometer spatial resolution, but simultaneous visualization of multiple antigens in parallel is an unresolved challenge. Herein, we demonstrate a DNA nanotechnology-based approach to label antigens in transmission electron microscopy images of tissue sections with high contrast patterns. For this, we attached gold nanoparticles to designated binding positions on DNA origami structures that act as visual "barcodes." These barcodes are then hybridized to complementary strands of DNA-modified antibodies that are bound to their respective antigens on ultrathin tissue resin sections. As a proof of concept, we demonstrate several types of barcodes and two different antibody labeling techniques that will expand the multiplexing abilities of immunostaining in a highly modular way.
