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Applications - General

1.1 Will my cells work with your systems?

In most cases, yes, your cells will work with the system. Whether a cell is compatible or not is dependent on many things such as the size and fragility of the cells. Please contact our applications team for a detailed list of cells that have been trialed in our Nadia product family or µEncapsulator system.

1.2 What size cells can be used in your systems?

The droplet junction size of the chip tends to dictate the size cells that can enter the system. We advise that cells are half the size of the junction, to avoid any sheering of cells and blockages. On the Nadia Instrument the maximum are cells of 50µm and on the µEncapsulator system a maximum of 25µm.

1.3 Can I use a very small sample of cells?

Yes. The Nadia Instrument and uEncapsulator system support volumes as small as 100µl.

1.4 What concentration of cells/beads do you need to run the scRNA-Seq application on the Nadia instrument?

Cells and beads are advised to be used at a concentration of 300 and 600 per µl, respectively.

 

1.5 Do you have a microfluidics chip capable of sorting droplets or large cells based on fluorescence?

Currently the most practical way of doing this is making small double emulsion droplets (an aqueous droplet in an oil shell, with an aqueous continuous phase), that can be sorted on a conventional FACS (https://www.dolomite-bio.com/applications/droplets-facs-sorting-libraries/).

1.6 What happens if you get gelled agarose in the chip?

To get this out, place the droplet chip on a hot plate with the gasket facing away from the metal surface at 90˚C for 5min, afterwards place the chip back into the µEncapsulator and continue with the cleaning protocol.

1.7 Are there any publications that show your products in use?

Yes, please see https://www.dolomite-bio.com/support/publications/ for current publications.

1.8 Do you sell beads/buffers/reagent kits?

At present, we do not sell beads.

1.9 Do you sell fluorinated oil and fluorinated surfactants?

Yes, we sell the Picosurf range of products.

1.10 What oil should I be using?
  • Pico-Surf™ 1 (In Novec7500)- Great for PCR. Stable, chemically inert droplets for PCR.
  • BioRad Droplet Generation Oil For EvaGreen – Suitable for RNA-Seq applications
1.11 Do you have a list of cell types that your system and Pico-Surf have been used with
  • Pico-Surf 1 5% in Novec 7500 has been successfully used with:
  • Mouse hybridoma
  • CHO
  • Human and Mouse B-cells
  • Human T-cells
  • Jurkat cells
  • HEK
  • 3T3
  • Endometrium
  • Annelid worm cells
  • Yeast
  • Plant cells such as corn
  • Bacterial cells such as E.coli and streptomycetes
  • Nuclei
  • Genomic DNA
1.12 Why are there different chips for RNA-seq applications on the Nadia instrument compared to µEncapsulator (T/B cell studies)?

This is because different types of beads are used in each application. RNA-Seq requires beads which are barcoded and at present are a size of 20-40µm. These beads therefore only run in a droplet chip with a 100µm junction as available for the Nadia Instrument and the scRNA-seq modular system. B and T cell studies require beads which don’t have a barcode and therefore are a size of 1 - 10 µm, which are able to run in the uEncapsulator 30/50µm chip.

1.13 What range of droplet sizes can you make on one chip?

Typically, 20% larger or smaller than the junction size (this is achieved by varying the pump pressure/flow rate).

1.14 What range of droplet sizes can you make on one chip?

Typically 20% larger or smaller than the junction size (this is achieved by varying the pump pressure/flow rate).

1.15 How long are droplets stable for?

Around a week at room temperature or two weeks at 4˚C.

1.16 Will the Dolomite Bio systems carry out downstream processes such as PCR/Sequencing?

No, the Dolomite Bio systems provide a solution for encapsulating single cells into droplets with/without beads. The protocol ends with a set of beads with mRNA attached, ready for downstream processing. If you are interested in data downstream of the encapsulation process please contact support@dolomite-bio.com.

1.17 Can you change buffers and reagents that are used in the system?

Yes, the Nadia Instrument when combined with Nadia Innovate and the µEncapsulator system are fully flexible allowing the use of own reagents and the development of applications.

1.18 Are our systems compatible with Illumina sequencers?

Yes

1.19 What is the maximum frame rate that you can capture per second?

4000

1.20 What is the minimum exposure time on the microscope?

0.05ms

Applications – RNA-Seq

2.1 What is the recommended concentrations of beads/cells in the RNA-Seq system?

We recommended 300 cells/ µl and 600 beads /µl, this reduces the chance of doublets of either a cells or beads. The system can be manipulated to higher concentrations to increase the chance of 1 bead and cell per droplet.

2.2 What is the capture efficiency of cells?

This is dependent on the concentration of the beads you are using, but when using our advised concentrations, the capture rate for cells is 10%. Meaning that 10% of the input cells will be captured, e.g. if you encapsulate 75,000 cells 7,500 will be captured for sequencing.

2.3 How many samples can I run in parallel

You can run either 1, 2, 4 or up to 8 samples in parallel in a 20min run.

2.4 Can I change droplets size or buffer compositions on the Nadia Instrument?

Run parameters such as pressures, stirrer speeds and temperatures can be altered when combing the Nadia Instrument with Nadia Innovate. This unlocks the Nadia Instrument for different applications or the use of different buffer compositions for scRNA-seq.

2.5 Why can't I run the RNA-Seq protocol in the µEncapsulator?

This is due to the junction size of the µEncapsulator chips and the properties of the barcoded mRNA capture beads. The Nadia instrument uses a 100µm chip, and the beads mentioned in the Macosko et al paper. These beads vary in size between 20-40 µm, so are compatible with the Nadia chips as their size is able to flow through the junction. However, the µEncapsulator has a junction size of either 15µm, 30µm or 50µm. This means that the beads are the size of the largest junction size, and wouldn't flow through, without blocking.

 

2.6 How much oil/cells/beads do you consume in one standard scRNA-Seq run?

250µl beads, 250µl cells and 3ml oil (for 20 min run and highest recommended flow rates).

2.7 How many single cell libraries can you make per hour?

Up to 48,000 single cell libraries in 20 minutes when using an 8-way cartridge on the Nadia instrument.

 

Product demonstration/workshops

3.1 How can I see your products?

If you are looking to see the products live and running, we regularly run workshops in many countries around the world.

3.2 When is the next workshop in my area?

Keep up-to-date with workshops being run near you by visiting the Workshops page.

Purchasing, installation and post-sale support

4.1 Do you provide installation and training?

Yes, installation and training can be (and typically for all system purchases, will be) included in the quotation. Please check your quotation.

4.2 Why do I need/what is installation and training?

Installation and training involves a specialist from the Dolomite Bio team visiting your institute to firstly install (set up and test performance of) the system. It also includes dedicated training of you and your colleagues, where we can help you understand the system, how to set it up and give guidance on your application. This often proves essential for users who have not used a microfluidic platform or are unfamiliar with the application and its set up.

4.3 How do I place an order?

If you already have a quote, please email your purchase order to orders@blacktrace.com (Dolomite Bio is a brand of Blacktrace). If you would like a quote, please visit Request a Quote.

4.4 What if I have a problem?

If you run into any problems with the system, please contact our support team at support@dolomite-bio.com

4.5 How do I get software updates?

The latest version of FCC is downloadable - Please contact support@dolomite-bio.com

 

Products

5.1 Are the chips consumables?

Nadia instrument and Nadia Innovate chips are consumables that are discarded after a run. The µencapsulator system uses glass chips, that when taken care of by using the cleaning protocol after every use, the chips can be reused many times.

5.2 How do I clean my Nadia instrument/Innovate?

The Nadia product family does not have wetted parts and therefore doesn't require cleaning. Chips are one time use and are therefore disposed of after every experiment.

5.3 How do I clean my µEncapsulator system/chips?

Normally the chips and system are cleaned by flushing through with 1% detergent, 100% isopropanol and then air (to dry the system/chip). Contact support@Dolomite-bio.com for a detailed cleaning protocol.

5.4 How do I know what products/consumables/accessories I need?

We are able to advise you and produce system quotes on products if you are unsure of what you need to run your application. Please contact us.

Dolomite Bio

6.1 What is the difference between Dolomite and Dolomite Bio?

Dolomite provides microfluidic components, such as pumps, chips and other accessories, which can be used for various different applications or function as microfluidic system solutions.

Dolomite Bio is focused on developing and selling products, typically complete systems and consumables to biologists. Many of these products are focussed on specific biological technologies such as Single Cell RNA Seq. Some of these products contain microfluidics but this is not the focus of Dolomite Bio.

6.2 Do you have any job vacancies at Dolomite Bio?

Yes, vacancies are updated regularly on our Careers