1. What type of cells can be used with the system?
You can use the QIAscout with a variety of eukaryotic cell types, including adherent or suspension cells, primary cells or cell lines, and fluorescent cells.
The QIAscout array is comparable to a standard cell culture dish – meaning you can plate and cultivate your cells in the same manner as a standard cell culture dish. You can seed both adherent and non-adherent cells, but since all cells need to actively adhere to the microraft, you should first coat the array with an adhesive solution if using suspension cells. Adherent cells, on the other hand, will stick to the array without the need for coating.
- 2. Will the QIAscout also work for circulating tumor cells (CTCs)?
Yes, if you are working with CTCs you can use the QIAscout for selection and isolation. We recommend first enriching the CTCs using the AdnaTest, and then coating the array to allow the CTCs to adhere – this is critical due to the fact that CTCs are often semi- or non-adherent. We recommend using between 4,000–6,000 cells on the array, so an enrichment step is a prerequisite for CTCs.
3. How long does it take to isolate a single cell?
Prior to using the QIAscout, cells need to be plated and then adhered to the array. The adherence step will vary depending on the cell line, but a period of 6–10 hours is generally adequate. Since the QIAscout can also be used for clonal expansion, you can cultivate cells for several days before isolating clonal colonies.
Once cells are attached and the QIAscout array is installed on your inverted microscope, isolating and transferring your cell of interest takes only about 1 minute per cell.
4. At what concentration should cells be applied to the array?
The goal is to maximize the number of microrafts with a single cell and minimize the number of microrafts with no cell or multiple cells. To do this, we suggest plating cells at a cell-to-microraft ratio of 1:2 or 1:3, as the cell distribution on the array follows a Poisson distribution. Since there are 12,000 individual microrafts, it is recommended to plate 4,000–6,000 cells. This will result in about 24–30% of the microrafts containing a single cell. A higher or lower number of cells will also influence the number of rafts having more than one or no cell.
5. What type of downstream applications can I use the single cells for after isolating with the QIAscout?
Once you have isolated single cells using the QIAscout, they can be used for a variety of applications, including single cell DNAseq, RNAseq, miRNA expression profiling, and whole genome amplification or whole transcriptome amplification for microarray, aCGH, PCR or NGS analysis. Since single cells are viable, they can also be used for further cultivation by transferring to a cell culture dish.
6. Is there any data available on whether QIAscout-isolated cells are pure and viable?
Yes, we recently published an application note on the viability and purity of single cells isolated using the QIAscout. Having live cells that are free of any contamination, such as from cell-free nucleic acids, is essential to be confident in your downstream DNA or RNA analyses. We found that cells isolated using the QIAscout system were both viable and showed no traces of contamination, making them ideal for further analysis.
7. How do I remove the cell from the raft when I’m ready for single cell analysis?
You don’t. The microraft can remain in the tube or well and will not affect downstream analysis.
8. Which microscopes and objectives are compatible with the QIAscout?
You will need an inverted microscope to use the QIAscout. The system has been verified to be compatible with Zeiss® microscope models Primovert, Axio Vert.A1 and Axio Observer, as well as selected models from Leica® and Nikon®, and works with most 4x, 5x and 10x objectives. You can find more detailed information on the compatible microscopes and objective dimensions in the QIAscout User Manual, “Appendix E – List of Microscopes and Objectives.”
9. What consumables do I need to order with the QIAscout?
The main consumable for the QIAscout will be the arrays, which contain 12,000 microrafts. Each array can only be plated once; however, it can be pierced as many times as you like, depending on how many single cells you want to isolate.
10. If the needle is somehow damaged, can it be replaced or would a new unit need to be purchased?
If the needle is damaged, it can easily be replaced by three screws. Two replacement needles are included with the QIAscout, and any additional needles can be ordered as a spare part/consumable. Please contact QIAGEN Technical Support or QIAGEN Customer Care to order spare parts.
11. Where do I learn more about QIAscout components and installation?
If you’d like to see what the QIAscout system constitutes and learn how it works, please watch a virtual demo here.
You can also watch a quick installation video on how to set up the QIAscout in your lab.
12. Is the QIAscout available in my country and how much does it cost?
Since this will vary by region, you can find more information on pricing for the QIAscout here.
Have any additional questions about the QIAscout? Comment below or contact us!