dSTORM Imaging Buffer Preparation

1. Thaw one vial of dSTORM Imaging Buffer Part A at room temperature for 10 minutes
2. Spin the reagent down using a tabletop centrifuge for 10 seconds
3. Carefully mix the solution with a pipette, making sure not to introduce bubbles

Note: Never flick or vortex dSTORM Imaging Buffer. Introducing oxygen will affect its performance

4. For each well to be imaged, add 99 µL dSTORM Imaging Buffer Part A to a new microtube

Note: We recommend imaging one well at a time, unless imaging time is estimated to be shorter than one hour

5. Remove dSTORM Imaging Buffer Part B from -20°C and let it come to room temperature. Ensure you re-seal the plastic bag it came out of
6. Add 25 µL of Part B Resuspension Buffer to the vial. Slowly add a single drop to the center of the pellet, and apply the rest to the sides of the vial to ensure complete dissolution of the pellet.
7. Put the rehydrated vial on ice for 5 minutes.
8. Gently mix the re-hydrated pellet by pipetting up and down. The vial can be removed from the ice bucket for this process. Do not introduce bubbles or air. Do not shake or vortex the vial.
9. Take 1 µL of the resuspended dSTORM Imaging Buffer Part B and add it to the 99 µL of dSTORM Imaging Buffer Part A in the fresh microtube. Once the required volume is removed, put the remainder at -20°C and use within 3 months
10. Remove Wash Buffer from wells
11. Add 100 µL of dSTORM Imaging Buffer mix to the well you are about to acquire. Note that we recommend adding dSTORM Imaging Buffer mix and imaging one well at a time, unless imaging times are under 1 hour
12. When preparing dSTORM Imaging Buffer sequentially, dSTORM Imaging Buffer Part A can be kept on ice during this time. dSTORM Imaging Buffer Part B should always be kept at -20°C and only taken out when needed