Epithelial Biology Center (EBC)

1. Coverslip placement: The coverslip should be placed such that it aligns as low (farthest from the header) on the slide as possible. The ideal location for a 24x50 mm coverslip is between 22 and 25 mm from the top of the slide (see example).
   
   Placement of the coverslip over the header will cause it to lay down at an angle. This will result in focus issues when doing microscopy and the pick-up arm of the Ariol to drop the slide. In addition, air will begin to infiltrate the mounting media at the site of offset with the header, shortening the lifespan of your slide.



2. Cleanliness: The cleaner your slide and coverslip, the cleaner the images you can collect from them. Simply put: garbage in, garbage out. Mounting media and fingerprints on the surface of the coverslip or the back surface of the slide can cause blurring, focus issues, and generally dirty appearance in your images. Slides should be cleaned prior to submission.
   
   For Immunohistochemistry, a clean, fresh razor blade should be used to remove any surface material from glass coverslips and the back of the slide. Isopropanol is used to remove figerprints and mounting media residue. For immunofluorescent slides, only clean water should be used to remover excess mounting media.

Improperly mounted slides must be hand-loaded for scanning (versus the automated robotic arm). They cannot be run as a batch, and thus other slides cannot be run in batch mode either. This slows the entire scanning process down for all slides and is an inconvenience for all users.

Due to this, slides that are improperly mounted may be rejected for scanning, or they may be scanned at the lowest priority and will take significantly longer to complete.

If you have any questions about mounting coverslips or cleaning slides, please contact me. If is best for everyone if we have these sort of things figured out before you mount or submit your slides.

Joseph Roland, Ph.D.
email (joseph.t.roland@vanderbilt.edu) or call (615) 322-0686