Hi all, I've been using the brand: Costar, for a 96-well microtiter plate.
Used all columns and rows, and yes the wells of the plate near the edge have lesser volume compared to the wells going to the middle.
How can I prevent this?
1. I don't stack plates.
2. The incubator is not crowded.
3. I wrap up the edges of the plate with parafilm and wrap the whole plate with aluminum foil.
Thanks in advance for the comments.
Dear Jonathan Tordilla
You are doing a good job. But more needs to be done.
Allow the cells to settle in the microplate while the microplate is on the benchtop. Around 15 to 60 minutes of settling time before the microplate is placed in an incubator is reported to reduce the “edge effect”.
The room temperature pre-incubation reduces the “edge effect” by minimizing thermal gradient in the edge wells of the plate during the period between dispensing of the cell suspension into the wells and subsequent settling of cells to the bottom of the well and adhesion to the surface. Thermal gradient is greatest in the edge wells of the plate, and it results in uneven distribution of cells on the well bottoms.
Secondly, minimize entrance to incubator to help reduce fluctuations in the internal environment. Consistent temperature and humidity level are necessary to avoid evaporation and the formation of condensation. Local convection currents of heat and gas coupled with the location of microplate within the incubator are among the most influential factors which you should consider.
Some more helpful hints to manage the “edge effect” may be obtained from the links provided below.
https://iccb.med.harvard.edu/sites/g/files/omnuum5406/files/iccb/files/an_hintsedgeeffecthts.pdf
https://bookcafe.yuntsg.com/ueditor/jsp/upload/file/20211020/1634697295188080171.pdf
Best.
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