For optimal performance the plate should be read within 60 minutes after the completion of the conjugate incubation as outlined in the package insert.

If the AtheNA Multi-Lyte^® batch name and the AtheNA work list name do not match exactly when entered, the analyze button will not turn green. It is critical that every time a batch name is entered into the AtheNA software, that the exact same name is also used in the AtheNA software. (Please refer to ZEUS Scientific Tech Tip 19)

Dilutions should be utilized immediately after processing. Failure to do so can lead to sample dilution evaporation which can skew the dilution concentration, which in turn affect results.

In order for results to be valid, the dd temp in the AtheNA Multi-Lyte^® software must be within the optimal range. This is indicated on the AtheNA "Run Batch Screen" as the area that is between the two yellow arrows and is highlighted in green. The dd temp is determined by the room or instrument temperature the last time the AtheNA was calibrated and if it deviates +/- 2 degrees while reading it will flag as out. Note that this will not be documented on the AtheNA results print out, therefore it is important to view the AtheNA screen while the run is in process.

In order to adjust probe height, the same filter plate type that is used for performing testing must be used with two large, flat, round disks in it.

The AtheNA Multi-Lyte^® instrument should be calibrated once a week, as part of the instrument's weekly maintenance procedure. Additionally, any time the dd temp goes out of range or the instrument is shut off or moved, the instrument should be re-calibrated.

If calibration fails, repeat again. Things to try are:

1. Check to see if your AtheNA Multi-Lyte^® probe is clogged,
2. Verify that the probe is set at the appropriate height,
3. Verify that the calibration beads were properly vortexed and sonicated,
4. Verify that you have placed sufficient volume of the beads in each well,
5. Be sure the correct bead values have been input into the correct locations,
6. Perform a weekly maintenance procedure on the instrument as this may be a sign that the instrument is not being properly maintained.

If after all these points, the calibration still fails, then call your AtheNA Multi-Lyte^® distributor to have an engineer come to assess your instrument.

All sonicators have different optimal water levels. To determine the optimal water level, water can be continuously added and removed until there is a visible area in the center of the sonicator where the water appears to splash up and be agitated. When a bottle of bead suspension is held in this "sweet spot", there should be an audible difference in sound intensity. Also, when the bottle is in this area, the laboratory technician should feel that it is the area of highest intensity. Only after verifying all these points can the water level in the sonicator be considered optimal.

Note: This optimal level may not be at the line inside the sonicator that has been painted there by the manufacturer. In internal testing at ZEUS Scientific, the optimal water level is normally well below this line.

While washing a filter plate, any unused wells should be covered by either parafilm, tape, or the technician's gloved hand. The vacuum pressure should not be turned off until wells have been completely evacuated of all liquid.

Note: the filter paper in the wells may still appear slightly wet, but any actual liquid must be evacuated. After the three washes, the filter plate should be gently blotted on a stack of paper towels until there is no more liquid being blotted out onto the towels. The pate should then be left on a non-absorbent surface for 3-5 minutes to air dry before proceeding to the conjugate step.

An ELISA plate should only be used if the desiccant inside the plate pouch is blue. A pink desiccant indicates that moisture has somehow entered the pate pouch, and this could affect results. To properly store an opened plate pouch to minimize this, follow the instructions listed in the packing insert listed under "Storage Conditions."