Product Support & Resources

Frequently Asked Questions

Aco-Dyes are best stored at 4°C, protected from light, for up to 6 weeks. Our 500-test kit format is supplied in multiple tubes to ensure long-term stability. We store them in ultra-pure sterile water for long-term storage to prevent contamination.

Can I freeze Aco-Dyes for long-term storage?

We do not recommend freeze-thaw cycles. Users have reported successful staining after 3 months of storage at 4C however we recommend using a stock solution up to 6 weeks in order to ensure optimal performance. 

What buffers can I use to reconstitute the Aco-Dye?

We recommend using Milli-Q water or standard formulations of PBS or TAE without additives like BSA or detergents.

How much Aco-Dye should I use for staining compared to the number of EVs?

We recommend performing a titration in your initial experiments to determine the optimal Aco-Dye concentrations for your EV samples. However, as a starting point: For 1e9 particles/mL, we suggest using a final Aco-Dye concentration ranging from 0.1 to 1 µM.

For 1e10 particles/mL, we recommend a concentration range of 1 to 10 µM.

Keep in mind that for every 10x increase in the concentration of particles, the concentration of dyes should also increase by 10X.

How long should I stain with Aco-Dye?

We recommend staining for 30 minutes to 1 hour at room temperature or 37°C. Staining at 4°C is suitable for longer incubations if desired.

Can I co-stain with antibodies or other dyes?

Yes, we recommend staining with Aco-Dye as the last in your protocol.

How can I remove excess Aco-Dye?

You can use an Amicon Ultrafiltration tube or dialysis; both methods work well with a 100 kDa MWCO.

Is sonication really necessary?

Sonication is highly recommended for nano-based experiments to ensure the Aco-Dye is fully solubilized. If you don’t have access to a sonicator, vortex the solution and briefly centrifuge, then repeat as needed.

Have the Aco-Dyes ever been used in in vivo models?

Yes, please check out our publications for further details.

What other solvents are the Aco-Dyes soluble in?

Aco-Dyes are soluble in DMSO, ethanol, and methanol.

Will Aco-490 work with a blue laser (488 nm)?

No, Aco-490 will not be excited at 488 nm. It is suited for use on a violet laser, 405 nm.

My staining seems uneven; how can I fix it?

It is crucial to add the EVs to the tube with Aco-Dye and pipet rapidly. Avoid adding the Aco-Dye to the tube of the EVs.  The initial staining interaction is very fast and adding the Aco-Dye to the EV solution can result in uneven staining.

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