Multiplexed bio-imaging using cadmium telluride quantum dots synthesized by mathematically derived process parameters in a continuous flow active microreactor

Quantum dots (QDs) are semiconductor nanocrystals with unique size-tunable emissions. To obtain a precise emission spectrum, monodispersity in size is imperative, which is achieved by controlling the reaction kinetics in a continuous flow of active microreactors. Further, a multivariate approach (dimensional analysis) is employed to impose stringent control on the reaction process resulting in monodispersed preparation of cadmium telluride (CdTe) quantum dots. Dimensional analysis knits multiple variables into a dimensionless mathematical form which not only predicts parameters precisely to obtain narrow size tunability but also guarantees reproducibility in synthesis. Analytical, structural, and optical characterization of the microreactor synthesized polydimethylsiloxane (PDMS) coated CdTe QDs reveal quantum efficient (61.5%), photostable (44%), and biocompatible nanocrystals of 5-15 nm. Further, PDMS-coated QDs (P-QDs) are conjugated with organelle-specific antibodies/biomarkers for in-vitro imaging in NIH 3T3 cells. Likewise, proliferating cell nuclear antigen (PCNA) and anti-myosin (MF20), cardiomyocytes antibodies are conjugated with P-QDs (red and green, respectively) to image the zebrafish's cardiac tissue. Antibodies tagged with quantum dots are imaged simultaneously using confocal microscopy. Thus, multiplexed bio-imaging of in-vitro and zebrafish tissue is demonstrated successfully. The results indicate the suitability of continuous flow active microreactor in conjunction with the mathematical prediction of process parameters to synthesize reproducibly monodispersed and quantum efficient QDs.