Posts in NanoSurface Plates
What protocols do you have for the use of the dishes. In the literature, several different seeding densities were used. What do you recommend?

As with any cultureware, optimal seeding density on ANFS can vary according to cell type and the purpose of the experiment. However, we recommend researchers use exactly the same cell density that they use for normal 35mm dishes, 6-well dishes and so on. Protocol information is provided on the ANFS product pages.

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Is there any type of pre-wash that must be performed prior to seeding the dishes?

No, ANFS is already gamma sterilized and plasma treated to promoted cellular adhesion. However, researchers may choose to deposit additional surface treatments depending on their exact cell type. Fibronectin, collagen, and PLL are all common surface treatment choices.

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Can I grow cells directly on NanoSurface Plates or do I need to do ECM protein coating?

We recommend coating the surface with the extracellular matrix protein you typically use in your cell culture workflow (collagen, fibronectin, gelatin, Matrigel, etc.). We also have a suggested concentration in our manual for protein coating of our substrates using some of these proteins. Before starting your experiments, make sure to check our manuals where you can find the details of the protocols for each protein coating.

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The coverglass comes in two different topography. Which one should I use for my application?

Our coverglasses come in two different formats: (1) flat and (2) NanoSurface substrates. Generally speaking, both formats have a polymer substrate that is softer compared to conventional coverglass. As a result, the softer coverglasses are more representative of the microenvironment of tissue with respect to the mechanical properties of the substrate. The NanoSurface substrate is featured with submicron grooves with the feature size comparable to that of collagen microenvironment in body. The pattern is also optimized to be in the size of the focal adhesion of cells and facilitate the alignment of adherent cells. Our customers usually use flat substrates either for their improved mechanical properties (softer substrate) or as a control sample for nanopatterned topography to show the differences in the functionality of aligned and non-aligned tissue. The patterned feature is specifically very useful for mimicking the microenvironment of many cell lines (such as pluripotent stem cells, cardiomyocytes, neurons, vascular muscle, epithelial, endothelial, mesenchymal stem cells, cancer cells and many other cell types) in vitro.

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What is the substrate thickness on NanoSurface plates and is the substrate transparent?

The substrate thickness (glass plus polymer coating) is specified at 160 - 190 microns (0.16 - 0.19 mm). This is the same thickness specification as a No. 1.5 coverglass. The polymer has an index of refraction comparable to that of glass and thus, is optically clear.

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