Trifunctional Linkers Enable Improved Visualization of Actin by Expansion Microscopy.
ACS Nano
; 17(20): 20589-20600, 2023 10 24.
Article
in En
| MEDLINE
| ID: mdl-37787755
ABSTRACT
Expansion microscopy (ExM) revolutionized the field of super-resolution microscopy by allowing for subdiffraction resolution fluorescence imaging on standard fluorescence microscopes. However, it has been found that it is hard to visualize actin filaments efficiently using ExM. To improve actin imaging, multifunctional molecules have been designed with moderate success. Here, we present optimized methods for phalloidin conjugate grafting that have a high efficiency for both cellular and tissue samples. Our optimized strategy improves anchoring and signal retention by â¼10 times. We demonstrate the potential of optimized trifunctional linkers (TRITON) for actin imaging in combination with immunolabeling using different ExM protocols. 10X ExM of actin labeled with optimized TRITON enabled us to visualize the periodicity of actin rings in cultured hippocampal neurons and brain slices by Airyscan confocal microscopy. Thus, TRITON linkers provide an efficient grafting method, especially in cases in which the concentration of target-bound monomers is insufficient for high-quality ExM.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Actin Cytoskeleton
/
Actins
Language:
En
Journal:
ACS Nano
Year:
2023
Document type:
Article