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4. If possible, the prearranged DIVA settings for Amplitude, Frequency, Drop 1 and Gap

should be used with Sweet Spot Monitoring to guarantee a stable stream.

5. An ideal stream will:

a. Be vertical and symmetrical with no slant or severe skew on droplets.

b. Have fewer than 6 satellite droplets.

c. Display fast-converging droplets on the 70um or 100um tip.

d. Stabilize with Sweet Spot on. Note that Amplitude and Frequency are locked

when Sweet Spot is on.

6. If the prearranged settings do not yield an ideal stream, begin with adjusting

Frequency. Frequency should be ~85-96kHz for the 70um and ~35-48kHz for the

100um. Use the highest value in that range that yields good droplet formation with a

low Amplitude value.

7. Adjust Amplitude (usually between 10-50) so there are at least 1-2 drops above and

below the break off point. Sweet Spot Monitoring tends to increase amplitude over

time, so starting with a lower value is best.

8. Set the gap target which yields the most distinct and cleanest side streams. Typically,

this is set to ~6. You can test different gap settings by adjusting amplitude.

9. Set the Drop 1 target based on the actual value displayed to the right. This serves as

a target for the Sweet Spot Monitoring, but also tracks drift over the course of the

experiment. If Drop 1 drifts +/- 10 from the original value, redo drop delay.

10. Activate Sweet Spot Monitoring. This will track drift in the Gap and Drop 1

measurements and automatically adjust Amplitude to compensate and return the

stream to your target values. It will also pause sorting if there is significant deviation

from the target Gap and Drop 1 until it is able to return the stream to normal. This

will not pause acquisition, so it's best to stop acquisition manually in the event of

major stream instability.

Optimizing the sort stream

1. Get the brightest/sharpest image of the waste and side streams by adjusting the

silver screw next to the sorting chamber.

2. In a test sort, set the two inner side streams a deflection of ~40 and the outer

streams to ~70. Then select the 2

drop so there is the least amount of fanning and the streams are distinct and clean.

3. Bring all side streams except the inner left back to zero, as you will need this for

drop delay.

Drop Delay

1. Open the Drop Delay experiment.

drop value in the stream window. Adjust 2