Laminin Poly-D-lysine coating coverslips
Neuvitro Corporation owns the leading technology to provide professional coating with the broadest coverage of substrates, such as PDL, PLL, PLO, Collagen, Gelatin, Laminin, Fibronectin, PDL+Laminin, PDL + Fibronectin, PLO+Laminin, PLO+Fibronectin, PDL+Laminin+Fibronectin, etc. The majority of coated German coverslips available from Electron Microscopy Science, ThermoFisher Scientific, and VWR is actually manufactured by Neuvitro Corporation. Free samples are available.
Laminin in cell culture:
As one of the major components of extracellular matrix, ECM, laminin have been used to enhance mammalian cell culture, especially in the case of culturing stem cells, as well as some primary cell cultures, which can be difficult to propagate on other substrates. Two types of naturally-sourced laminins are commercially available. Laminin extracted from mouse sarcomas is one popular laminin type, another is laminin mixtures from human placenta.
Problem of coating laminin over glass surface:
Unfortunately, laminin does not like to stay on surface of glass coverslips. Laminin molecules comes off quickly during coating procedure.
Poly-D-lysine (PDL) helps the link between laminin and glass and solve the problem.
Now Laminin poly-D-lysine coated coverslips are available.
How much laminin needed to coat my dish / plates / coverslips?
Minimum required amount of laminin is 0.05ug/cm² surface area of your culture dish or coverslips. For example, to coat 100 pieces of 18mm diameter coverslips, the total surface area of 100 pieces of coverslips is 0.9 x 0.9 x 3.1416 x 2 x100 = 510cm². The minimum required laminin is 25.5ug.
How to know if my laminin coating is good or not?
It has been well known that:
The data quality of cell based assays depends on the quality of cultured cells.
The quality of adherent cells in culture depends on the substrate cells attached to because 50% cell surfaces are occupied by the substrate. Cells interact each other through coated extracellular matrix (ECM). The extracellular matrix not only connects cells together, but also relay environmental signals to cells and guides cell movement and migration.
Therefore, you should pay detailed attention to your coating protocol and your selection of optimized coating molecule.
The following simple procedure will help you to determine the compatibility between your coating molecule and your cells:
Obtain a free package of Cell Culture Control Standard from Neuvitro Corporation. The Neuvitro control standard is digitally manufactured and tested, showing extremely high consistency, repeatability, and culture quality.
Have a multi-well culture plate and place both of your own coverslips and different kinds of Neuvitro control standards in comparison pairs into different wells.
Seed cells at reduced cell density into wells and reduced serum to 1%.
Culture the cells overnight and compare their growth in different wells. From this simple test, you will know:
If you have used the correct coating molecule.
If you have made good coating.
If your coating is not good, how much better you should anticipate during your future improvement.
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