Although we believe that the cell surface sialic acids (Sias) are playing an important role in cell-cell interactions and related tumor metastasis processes, acquisition of their quantitative information has yet been a challenge to date. Here, we reported the construction of a new analytical platform for Sias-specific imaging and quantification. We used N-azidoacetyl-mannosamine tetraacylated as a metabolic sugar substrate to bioassemble azido-Sias on the surface of cells via the metabolic pathway of Sias de novo synthesis. These azido-Sias allow us to perform a duplex Sias-specific analysis with various fluorescent and elemental reporters such as DIBO-Alexa Fluor 647, DBCO-DOTA-Eu, and DBCO-PEG-BODIPY, which can be easily labeled and/or tagged through an effective copper-free bioorthogonal click reaction. Compared to the previous reported strategies, we quantified the cell surface Sias with the LODs (3σ) down to 8.9 fmol and 0.24 pmol using Eu- andB-species unspecific isotope dilution ICPMS, in addition to their red- and green-CLSM profiling. Such a platform enables us to evaluate Sias regulation under the administration of paclitaxel, finding that 1 μM paclitaxel induced a significant Sias decrease of 67% on the surface of hepatic tumor cell SMMC-7721, while had no obvious adverse effect to that of para-carcinomatous liver cell LO2. Besides Sias, we believe that this metabolism-based click-mediated platform will provide opportunities to study other monosaccharides and their corresponding biological roles when more corresponding chemically modified sugar substrates and specific bioorthogonal reactions are developed.

Although rare cancerous cells are considered as more objective indications for a precise early diagnosis of cancers, accurate counting of them still is a spirited challenge. We reported a signal multiplication strategy by constructing element-tagged viruslike nanoparticles (VLNPs) with a precise number of atoms for a membrane biomarker mediated higher sensitive cell counting using inductively coupled plasma mass spectrometry (ICPMS). Typical bacteriophage MS2 was exemplified to demonstrate the effectiveness of the element-tagged VLNPs as signal multipliers. Dibenzylcyclooctyne-poly­(ethylene glycol)-folate (DBCO-PEG-FA) and DOTA-Eu complex tag modified (FA-PEG)69-MS2-(DOTA-Eu)965 targeted the folate receptor (FR) on KB cells as low as subzeptomole FRs could be quantified by 153Eu-species unspecific isotope dilution ICPMS, allowing us to be able to count at least 5 KB cells. While more than 2197 KB cells were needed to give a significant ICPMS signal using FA-PEG-DOTA-Eu, demonstrating more than 2 orders of magnitude signal multiplication and resulting in total 4.0 × 108 times signal amplification relative to one KB cell. We believe that such a signal multiplication strategy can be expanded to quantify and count other membrane biomarkers and their host cells using various VLNPs modified with different kinds and precise numbers of elements and guiding groups. In this way, prescribed multiples of signal amplification can be realized for a more accurate ICPMS-based quantitative bioanalysis because targeted molecules/cells in a complicated biological system might exist in orders of magnitude wide concentration range.