Glass Slide Microarray Technique Video
Hi researchers! This video is an overview of the best practices for handling RayBio glass-slide antibody and protein arrays. After removing your array kit from the freezer, let the slide equilibriate to room temperature for 20-30 minutes before opening the bag. After unwrapping, the slide should be completely air-dried before use, which usually takes 1-2 hours. This step is very important, as residual moisture can result in smearing of the antibody spots. Each slide comes attached to a removable chamber assembly and silicone gasket. Each well on the slide contains an identical array of antibody or protein spots. If you are using more than one slide, you should mark the slides to differentiate them. It’s best to make a very small marking along the top or bottom edge using a fine-point permanent marker. In our experience, red or black ink can interfere with the array signals. So only use blue or green ink on the slide. Never write on the actual well area, as this will interfere with scanning. When you are ready to being the experiment, be sure to orient the slide and record which sample or standard is being added to which well. Check you manual for suggested configuration of standards. To avoid smearing the array spots, pipette slowly and never touch the pipette tip to the glass. Be sure the liquid fully covers the array surface, with no air bubbles. Take care not to overfill the wells. This will not happen if you added the recommended volumes of liquid. All incubation steps, including washes, should be performed with gentle rocking or rotation. We recommend a half-cycle per second. Avoid vigorous shaking of the slide, which can cause splashing. We recommend incubation at room temperature. However, the sample incubation step may be performed overnight. Overnight incubation should be done at 4 degrees Celsius, with sealed wells avoid evaporation . The following day, allow the slide to warm up to room temperature for 1 hour before proceeding with the next step. Much like running an ELISA, good washing technique is critical. When aspirating the wells, tilt a slide and place the pipette tip in the lower right corner to ensure complete removal of the liquid without touching the array spots. To remove the chamber assembly, simply push the chips outward and carefully remove the incubation chamber and gasket. Take care to handle the slide only by the edges. Never touch the array surface and wear gloves to avoid fingerprints and dirt. After taking the slide apart, the final washes are performed by completely submerging the slide in wash buffer and gently shaking the tube. After the final wash, dry the slide by removing as many droplets as you can. The easiest way is using a pipette tip with gentle suction. Again, take care not to touch the antibody spots. After taking the slide apart, you’ll see a green marking on the lower right corner. This indicates the front side of the slide, or the array slide. For most array scanners the array side should be facing the laser. Always handle the naked slide by the edges only. If you wish to mark the slide at this point, use blue or green permanent ink. And keep the marking very small. Never ever write or touch the arrayed well areas on either side, as this will interfere with laser transmission. We also do not recommend using adhesive labels or tape on the slides. After scanning your slides, store them at room temperature, always away from light and dust. If you do this, the array signals will remain bright for over a year.