Tips include why wash dishes should be cleaned with appropriate solvent and are free of fluorescence, what can be caused by droplets of buffer still present on slide after washing, and how to ensure there is a good mixing of the wash solutions over the surface of the slides.
Ensure wash dishes are cleaned with appropriate solvent and are free of fluorescence. Washing should be done after every array processing run and monitoring the background signal intensity from each run will give an indication if there is an issue.
Mike Ornelas
FAS Manager – North America
The correct volume of wash buffer 2 and wash dish 2 should be prewarmed at 37°C, ideally overnight.
Pete Gray
Array and NGS Field Application Specialist, Europe
Droplets of buffer still present on slide after washing will result in artefacts on tiff image.
Mike Ornelas
FAS Manager – North America
Control ozone levels by washing slides in an ozone-free cabinet and avoid unnecessary exposure of slide to environment.
Pete Gray
Array and NGS Field Application Specialist, Europe
Remove slides slowly from final wash buffer to allow surface to dry completely. Droplets of buffer left on the surface of the slide will cause scanning artefacts that can impact on data quality.
Mike Ornelas
FAS Manager – North America
Washing should be performed on a magnetic stirrer producing a vortex in the wash buffers. Setting the stirring platform so that the rotation is just starting to produce a vortex will ensure there is good mixing of the wash solutions over the surface of the slides. Also, make sure the mixing is not too vigorous otherwise there is a risk the slides will be disturbed or the stirrer bar will jump and hit the slide surface.
Pete Gray
Array and NGS Field Application Specialist, Europe