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Activated plasmonic nanoaggregates for dark-field situ imaging for HER2 protein imaging on cells surface.

Dark-field microscopy (DFM) based on localized surface plasmon resonance (LSPR) was used for observation of experimental phenomena, which is a hopeful non-damaged and non-photo bleaching biological imaging technique. In this strategy, plasma nanoaggregates with stronger scattering efficiency were formed in the presence of the target, causing a "turn-on" phenomenon, when asymmetry modified AuNPs were introduced as probes with zero LSPR background. First, Au1-N3 probe and Au2-C≡C probe was designed for the cycloaddition between azide- and alkyne- to form AuNP dimers under the catalytic action by Cu+ which obtained from the reduction of Cu2+ by sodium ascorbate. The two kinds of probes were used for the detection of Cu2+ in rat serum successfully. Then, for applying this concept to protein on cells, DNA and antibody were modified on the probes. DNA1/Au1-N3 probe and anti-HER2/Au2-C≡C probe were proposed for HER2 protein DFM on cells. By designing aptamer sequence in primer, the rolling circle amplification (RCA) was introduced in HER2 DFM on cell, and the image signal was much brighter than no-RCA. The unique design made it was more easily to discriminate the target signal from background noise in cell DFM. This method might be used in the fields of molecular diagnostics and cell imaging.