Advanced Healthcare Materials recently published Phenotypic Assessment and Gene Expression Analysis from Live Cells in which addresses the Cells-to-cDNA on Chip by utilizing the I.DOT Liquid Handler enabling the precise handling of nanoliter volumes, facilitating advanced phenotypic assessments and gene expression analyses from live cells with unprecedented accuracy and efficiency (Fig. 1).
Figure 1. Screenshot of "Cells-to-cDNA on Chip: Phenotypic Assessment and Gene Expression Analysis from Live Cells in Nanoliter Volumes Using Droplet Microarrays," published in Advanced Healthcare Materials.
The researchers from the Karlsruhe Institute of Technology, Germany, discovered that integrating the I.DOT's non-contact technology into their workflow was a game changer. The precision and efficiency it provided were unparalleled, allowing them to achieve high-quality results quickly and with a minimal waste.
Additionally, the ability to handle live cells and reagents at such a small scale opened up new possibilities for their phenotypic assessments and gene expression analyses using the Droplet Microarrays (DMA). The authors are excited about the future applications of this technology in our customers' ongoing and future research projects using the I.DOT Non-Contact Dispenser.
The sample preparation workflow of cells-to-cDNA on chip would not have been efficiently or accurately carried out without the inclusion of the I.DOT Non-Contact Dispenser, which dispensed various reagents at different experimental steps in micro/nanoliter volumes onto the hydrophilic spots (for 1 mm DMA) (Fig. 2). These include a solution containing appropriate volumes of HeLa-CCL2 total RNA (DSMZ GmbH, Germany and BioCat GmbH, Germany), dNTPs and random hexamers/oligo dT primers and other reagents of reverse transcription (Table 1).
Figure 2. Workflow of “Cells-to-cDNA on Chip”. Briefly, 1. Cell culture and screening in micro/nanoliter droplets dispensed on DMA using the I.DOT Liquid Handler, 2. phenotypic analysis of cells by microscopy-based methods, 3. cell lysis and isolation of mRNA using poly-T magnetic beads, 4. mRNA to cDNA conversion within the same droplet, 5. Collection of cDNA for qualitative and quantitative gene expression analysis. Image modified from Chakraborty et al. (2022).
Table 1. Reagent volumes for cDNA synthesis on 1 mm DMA using the I.DOT Liquid Handler.
The "Cells-to-cDNA on Chip" workflow relies heavily on I.DOT for its efficiency and accuracy. The technology precisely dispenses various reagents in micro and nanoliter volumes throughout the process (Fig. 2). These include solutions containing RNA, primers, and other essential components for reverse transcription (Table 1).
The DISPENDIX team is enthusiastic about the potential of the I.DOT Non-Contact Dispenser. This technology has the power to revolutionize various fields:
The marriage of I.DOT technology with gene expression analysis marks a significant leap forward. By enabling the meticulous study of live cells, this innovation promises to:
Curious about how the I.DOT can elevate your research? Book a demo today and see this cutting-edge technology in action!
Chakraborty, S., Luchena, C., Elton, J. J., Schilling, M. P., Reischl, M., Roux, M., Levkin, P. A., & Popova, A. A. (2022). "Cells-to-cDNA on Chip": Phenotypic Assessment and Gene Expression Analysis from Live Cells in Nanoliter Volumes Using Droplet Microarrays. Advanced healthcare materials, 11(12), e2102493. https://doi.org/10.1002/adhm.202102493