====================== Attune troubleshooting ====================== The following troubleshooting tips and explanations are a result of empirical observations and discussions with our support engineers. Additional troubleshooting is available in the `Attune maintenance manual <../../_static/files/attune_maintenance_guide.pdf>`__. Delay before events start running --------------------------------- If you notice that there is a delay before events appear, but the events appear normal (e.g. there isn't a clog), check that the thumbscrew and screw at the top of the main sample syringe are fully tight. You can add about a quarter-turn if they are loose. **Explanation:** The main sample syringe is responsible for pushing liquid, including your sample, through the fluidics system. If the two screws that attach the servo motor to the syringe are loose, then when the servo motor initially starts moving, that motion instead goes towards the slop in the system and the main syringe doesn't start moving until after a delay. This delay actually affects any procedure that starts from the fully-down syringe position, but is most noticeable when capturing events. The delay is not harmful in any way. Fluid leak from the autosampler SIP onto a running plate -------------------------------------------------------- First, take the plate out and clean up the spilled liquid. Carefully, **while the Attune is idle or off**, you can open the autosampler door and clean up any spilled liquid with a paper towel. Then, ensure that the two waste bottle connections are fully connected. Make sure you re-seat the autosampler waste bottle connection unti you hear **two clicks**. **Explanation:** The autosampler fluidics are relatively simple. The autosampler syringe directly pulls focusing fluid from the autosampler focusing fluid bottle to use as sheath fluid. After aspirating a sample, the sample syringe is actually *pushing* fluid through the system to move your sample into position, with the excess fluid "in front" of your sample redirected into the waste bottle. If the autosampler waste bottle is not fully connected, you instead end up with the syringe trying to push against a closed connection, which leads to high backpressure. The fluid goes through the easiest escape route, which is back out of the SIP onto your plate :( Wells skipping and the autosampler fails to draw focusing fluid --------------------------------------------------------------- If you directly observe that fluid is not successfully being taken up by the autosampler syringe, or you observe that no events happen or wells get entirely skipped, the autosampler is likely not taking up focusing fluid correctly. Check that the autosampler focusing fluid bottle connection is fully seated. Also check that the autosampler waste bottle connection is fully seated (two clicks on insertion). **Explanation:** The syringe has to be able to hold suction. A leak on the focusing fluid bottle side will prevent the syringe from getting fluid in (unless you elevate the focusing fluid bottle). A not fully connected waste line will cause pressure to build up in the system and prevent it from working. Power cycling the Attune ------------------------ If the machine is behaving oddly in a way not described above, or if the software prompts you to do so, power cycle the machine. To turn the Attune off, switch both the autosampler and the main machine off, and let them sit for 30 seconds. Then, to turn the machine on, **first turn on the autosampler** (switch is labeled on top with a 1) then turn on the main machine (switch is labeled with a 2). **Explanation:** The autosampler should be turned on first so that the main machine is able to sense that it is connected and initiate the correct settings.