The Poly-L-Lysine coating expert

    neuVitro     

 Poly-Lysine coating cell culture 

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         neuVitro        

 The poly-lysine coating expert

 

Should I dissolve polylysine in dd-water for coating?

  • Poly lysine coating in dd-water is simple but the coating efficiency is low. The pH of dd-water is acidic, usually lower than pH6. After dissolving polylysine, the ph value is further reduced to 3, which is not optimized for poly lysine coating. 

  • Solution: Use high pH polylysine coating, such as pH 8.5, in borate buffer, which enhances poly lysine adsorption to cell culture surface. 

  • Critical: The borate buffer residue must be rinsed away thoroughly from coverslips before cell culture.

How much poly lysine needed to coat my dish / plates / coverslips?

  • High concentration polylysine is recommended. Minimum required amount of poly lysine is 3ug/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 required poly lysine is 510x3ug = 1530ug.

How to know if my polylysine 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 package of Cell Culture Control 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 a good coating protocol.

    • If your coating is not good, how much better you should anticipate during your future improvement.

       Table of polylysine needed for coating dishes, plates, and coverslips

Cell culture devices

Total coating area per plate

Polylysine needed

6 well culture plate 57cm² 427.5ug polylysine at 50ug/ml
12 well culture plate 45.6cm² 342ug polylysine at 50ug/ml
24 well culture plate 45.6cm² 342ug polylysine at 50ug/ml
48 well culture plate 45.6cm² 342ug polylysine at 50ug/ml
96 well culture plate 30.7cm² 230ug polylysine at 50ug/ml
35mm dish 9cm² 67.5ug polylysine at 50ug/ml
60mm dish 21cm² 157.5ug polylysine at 50ug/ml
100mm dish 55cm² 412.5ug polylysine at 75ug/ml
150mm dish 152cm² 1140ug polylysine at 75ug/ml
245mm dish 500cm² 3750ug polylysine at 100ug/ml
12 mm coverslips round 2.3cm² 17.25ug polylysine at 100ug/ml
15 mm coverslips round 3.5cm² 26.25ug polylysine at 100ug/ml
18 mm coverslips round 5cm² 37.50ug polylysine at 100ug/ml
22 mm coverslips round 7.6cm² 57ug polylysine at 100ug/ml
25 mm coverslips round 9.8cm² 73.50ug polylysine at 100ug/ml

 

 

Neuvitro Corporation

The coverslip coating expert  

 

 

with G3 Digital Coverslip Coating System

 

 

Poly-D-lysine coating

Pre-coated coverslips

Ready to use German coverslip with poly-D-lysine (PDL) coating on both sides for cell culture, starting $55/box

 

Rat tail Collagen

Pre-coated coverslips

Ready to use German coverslip with Millipore rat tail type I Collagen coating on both sides for cell culture, starting $65/box

 

Fibronectin coating

Pre-coated coverslips

Ready to use German coverslip with fibronectin coating on both sides for cell culture, starting $89/box

 

Laminin coating

Pre-coated coverslips

Ready to use German coverslip with human Laminin coating on both sides for cell culture and neuron culture, starting $94/box

 

Gelatin coating

Pre-coated coverslips

Ready to use German coverslip with porcine gelatin coating on both sides for cell culture, starting $89/box

 

German Coverslips 

Cell culture tested

CE certified, tested for primary cell and primary neuron cultures, starting $12/box

 

Liberty 120

High-speed gel system

See you PCR and genotyping results in 6 min with straight banding technology...

 

FAQ107