1. I find some areas within wells where there are no cells attached. How can I avoid this?
Use of a manual pipettor can sometimes result in the formation of air bubbles which will prevent cells from attaching to the microplate surface. Use an automated pipettor or liquid handler to reduce the chances of pipetting difficulties. You can also pre-wet the wells with small volumes of medium (25 µL per well for 96-well half area (A/2) microplates or 10 µL per well for 384-well microplates) and quickly centrifuge to ensure media reaches the bottom of all the wells. Cells can be manually seeded more easily immediately following this.
2. I noticed an uneven distribution of cells in the outer wells of my microplate. How do I reduce this edge effect?
After seeding the microplates, let them sit for 30 to 45 minutes in order for cells to settle prior to moving to the incubator. This will negate any effect from temperature differences from the edge of the microplate compared to the center of the microplate that occurs upon initial incubation.
3. Why are the wells round?
The round well geometry prevents wicking of liquid within the well and prevents cells from aggregating into square well corners.
4. What are the main dimensions of the microplate that I need in order to use Corning high content screening microplates with my in-house imager?
5. Why do the Corning high content screening microplates have smaller wells than other manufactures?
Corning 96-well half area (A/2) and 384-well microplates have smaller wells to allow for less use of reagents and cells, which may result in a cost savings. The 96- and 384-well bottom surface area is sufficient to capture quality data for high content imaging. Using a high content screening imager, we determined the number of fields of view that could be obtained without capturing the edge of the well. For most users, these fields of view are acceptable to capture more than 1,000 nuclei per well.
6. I seem to have greater evaporation along the edges of my microplate, and I worry that it could affect my assay. How can I reduce this type of edge effect?
Make sure there is sufficient media volume per well. Volumes of 40 µL per well in 384-well microplate and 100 µL per well in 96-well half area (A/2) microplate should show limited evaporation for a 24- to 48-hour culture period. For longer culture periods, breathable tape can significantly reduce evaporation without inhibiting gas exchange.