Multichannel Systems MEA Manual - Manual (Page 40)

MEA Manual
40
Procedure
1. Incubate the MEA with polyornithine solution at RT for 2 – 3 hours or overnight at 4 °C.
2. Aspirate the polyornithine solution and rinse the MEA 3 x with distilled water before direct use or before
the following coating with laminin. MEAs coated with polyornithine can be stored at 4 C for several
weeks.
3. Incubate pre-coated MEA with laminin solution for at least 1 h.
4. Aspirate the laminin solution and directly plate cells.
Literature
Cellular Neurobiology, A practical approach, ed. By Chad and Wheal, IRL Press, Oxford
5.4.4 Coating with Poly-D-Lysine (plus Laminin)
Poly-D-lysine has been used by several groups. Results seem to be equivalent to a coating with
polyornithine. Some users complained about cell clumping and resulting cell death when using
poly-D-lysine and had better results when using polyethylenimine (PEI).
Materials
 Poly-D-lysine 5 mg / 10 mL (= 0.05 % w/v) stock solution (Sigma-Aldrich, Inc., P7280)
 Laminin solution 1 mg/ml (Sigma-Aldrich, Inc., L2020)
Laminin solution
 20 μg/ml laminin in plating medium or PBS (phosphate buffered saline).
Procedure
1. Incubate the MEA with poly-D-lysine solution and incubate at 4 °C over night.
2. Rinse MEA with sterile distilled water 3x to remove toxic unbound lysine and let the MEAs air dry under
sterile conditions (laminar flow) before plating the cells, or before the following coating with laminin.
MEAs can be stored at 4 °C for up to two weeks.
3. Incubate pre-coated MEA with laminin solution at 4 °C over night.
4. Aspirate the laminin solution and directly plate the cells.
Literature
Goslin et al., 1988, Nature 336, 672-674
Maeda et al., 1995, J.Neurosci. 15, 6834-6845
Gross et al., 1997, Biosensors & Bioelectronics 12, 373-393