Troubleshooting

stage not connecting in nis-elements

Disconnect stage USB connection and power cycle and restart NIS-elements.

Whenever stage is power cycled - normally we leave the stage on - then follow these steps to initiate the stage.

pimag/dOPM refocussing actuator is not responding

Diagnose the problem:

1. remove dOPM enclosure lid and see if pimag is moving when controlling it in NIS-elements
2. run the Pikromove software and follow these steps
3. If Pikromove software cannot communicate with Pimag then power cycle the Pimag control box by pulling the cable out of the back and go back to step 2.

lasers not turning on

laser power seems to have dropped

nis-elements optical configurations not found

computer won't boot

strange background noise

alignment of dOPM optics not right

my prefind is not working

stage coordinates are not right

dOPM dual view bead data cannot be registered

my position list is not correct

Has the sample been displaced by contact with the microscope objective.

My sample is unhappy

• is my imaging media OK
• is the CO2 humidifier empty/topped up
• is the CO2 5% and stable - use meter
• is temperature stable
• is there an infection
• phototoxicity

A way to rule out dOPM is to run duplicate assays and keep in your incubator

Also beneficial to collect low res widefield images of wells to get an overview of typical growth