Multiplex cancer cell imaging with dna-paint (review article)

Nasrin Mohajeri,1 Mohammad pourhassan moghaddam,2 Abolfazl akbarzadeh,3 Nosratollah zarghami,4,*

1. Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences
2. Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences
3. Department of Medical Nanotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences
4. Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences

Abstract


Introduction

Super-resolution fluorescence microscopy is the main challenge in biological and biomedical enquiries. dna points accumulation for imaging in nanoscale topography (dna-paint) allows researchers to detect a large number of distinct target agents. dna-paint provides an excellent super-resolution imaging method that attains less than 10 nm spatial resolution in vitro and in situ using dna structure.

Methods

Dna origami formation was carried out in a one-pot reaction containing scaffold standard. dna paint docking strands need folding buffer with suitable ion concentration. for cell immunostaining used common cell line such as hela cell. to intracellular structure imaging should be used fluorescence microscopy with perfect focus system. transmission electron microscopy (tem) and atomic force microscopy were essential for supplementary studies. super-resolution data processing were reconstructed using fiducial-based, spot-finding and 2d-gaussian fitting algorithms.

Results

Researchers found that the spatial resolution of dna-paint depends on enhanced drift correction and advanced localization precision. multiplexed cellular imaging with dna-paint labeled protein targets using antibodies conjugated with dna docking strands. transient binding of dye-labeled dna strands to their complementary target sequence (docking site) attached to a molecule of interest. the transient binding of imager strands is detected as blinking.

Conclusion

Dna-paint provide a simple, easy and powerful method for highly super-resolution imaging. imaging groups researchers anticipate this method will become a standard and promising strategy for studying complex bimolecular systems. we can use this method in multiplex detection of the cancer cell with high resolution. although the only limitation is the orthogonality of dna-paint sequences.

Keywords

Dna-paint multiplex imaging