Sequential super-resolution imaging using DNA strand displacement.
Sequential labeling and imaging in fluorescence microscopy allows the imaging of multiple structures in the same cell using a single fluorophore species. In super-resolution applications, the optimal dye suited to the method can be chosen, the optical setup can be simpler and there are no chromatic...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2018-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC6118358?pdf=render |
| _version_ | 1811209009083973632 |
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| author | Sandeep Pallikkuth Cheyenne Martin Farzin Farzam Jeremy S Edwards Matthew R Lakin Diane S Lidke Keith A Lidke |
| author_facet | Sandeep Pallikkuth Cheyenne Martin Farzin Farzam Jeremy S Edwards Matthew R Lakin Diane S Lidke Keith A Lidke |
| author_sort | Sandeep Pallikkuth |
| collection | DOAJ |
| description | Sequential labeling and imaging in fluorescence microscopy allows the imaging of multiple structures in the same cell using a single fluorophore species. In super-resolution applications, the optimal dye suited to the method can be chosen, the optical setup can be simpler and there are no chromatic aberrations between images of different structures. We describe a method based on DNA strand displacement that can be used to quickly and easily perform the labeling and removal of the fluorophores during each sequence. Site-specific tags are conjugated with unique and orthogonal single stranded DNA. Labeling for a particular structure is achieved by hybridization of antibody-bound DNA with a complimentary dye-labeled strand. After imaging, the dye is removed using toehold-mediated strand displacement, in which an invader strand competes off the dye-labeled strand than can be subsequently washed away. Labeling and removal of each DNA-species requires only a few minutes. We demonstrate the concept using sequential dSTORM super-resolution for multiplex imaging of subcellular structures. |
| first_indexed | 2024-04-12T04:32:35Z |
| format | Article |
| id | doaj.art-9a90de041cce44a58fe82f4156bb06a3 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-12T04:32:35Z |
| publishDate | 2018-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-9a90de041cce44a58fe82f4156bb06a32022-12-22T03:47:54ZengPublic Library of Science (PLoS)PLoS ONE1932-62032018-01-01138e020329110.1371/journal.pone.0203291Sequential super-resolution imaging using DNA strand displacement.Sandeep PallikkuthCheyenne MartinFarzin FarzamJeremy S EdwardsMatthew R LakinDiane S LidkeKeith A LidkeSequential labeling and imaging in fluorescence microscopy allows the imaging of multiple structures in the same cell using a single fluorophore species. In super-resolution applications, the optimal dye suited to the method can be chosen, the optical setup can be simpler and there are no chromatic aberrations between images of different structures. We describe a method based on DNA strand displacement that can be used to quickly and easily perform the labeling and removal of the fluorophores during each sequence. Site-specific tags are conjugated with unique and orthogonal single stranded DNA. Labeling for a particular structure is achieved by hybridization of antibody-bound DNA with a complimentary dye-labeled strand. After imaging, the dye is removed using toehold-mediated strand displacement, in which an invader strand competes off the dye-labeled strand than can be subsequently washed away. Labeling and removal of each DNA-species requires only a few minutes. We demonstrate the concept using sequential dSTORM super-resolution for multiplex imaging of subcellular structures.http://europepmc.org/articles/PMC6118358?pdf=render |
| spellingShingle | Sandeep Pallikkuth Cheyenne Martin Farzin Farzam Jeremy S Edwards Matthew R Lakin Diane S Lidke Keith A Lidke Sequential super-resolution imaging using DNA strand displacement. PLoS ONE |
| title | Sequential super-resolution imaging using DNA strand displacement. |
| title_full | Sequential super-resolution imaging using DNA strand displacement. |
| title_fullStr | Sequential super-resolution imaging using DNA strand displacement. |
| title_full_unstemmed | Sequential super-resolution imaging using DNA strand displacement. |
| title_short | Sequential super-resolution imaging using DNA strand displacement. |
| title_sort | sequential super resolution imaging using dna strand displacement |
| url | http://europepmc.org/articles/PMC6118358?pdf=render |
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