FAQ - General questions

Why do you recommend that the Slide H is stored at -20°C? Is this just a precautionary measure or is it based on experiments?

This recommendation is based on long term stability / storage experiments. We have found that there is a slow hydrolysis of the NHS-esters over time, so to achieve a full 12-month shelf life, we recommend (long term) storage at -20°C. However, NEXTERION® Slide H can be shipped or stored at ambient temperatures for short periods of time (less than one month) without any significant hydrolysis.

What is the thickness and micro-roughness of the SCHOTT NEXTERION®
Slide H coating?

The coating on NEXTERION® Slide H has been measured using atomic force microscopy (AFM) and ellipsometry. As the coating swells in the presence of water it has been measured in both the dehydrated and hydrated state.
The dehydrated NEXTERION® Slide H coating is approximately 10 - 20 nm thick with a peak-to-trough roughness of less than 1 nm (rms roughness = 0.29 nm), similar to the surface of the underlying glass.
After 30 minutes hydration, the film thickness stabilizes at 50 - 100 nm, and has a final peak-to-trough roughness of approximately 10 - 20 nm (rms roughness = 3.20 nm).

What solvents can be used in my spotting buffer for printing on NEXTERION® Slide H?

Spotting buffers based on water are preferred. DMSO is not recommended as co-spotting reagent, but might work if the concentration is not too high. Glycerol is generally not recommended.

If I want to store my NEXTERION® Slide H slides after printing but before hybridization, what are the recommended conditions for protein and DNA arrays?

The optimal conditions would be to store the printed slides in the original 25-slide mailer boxes sealed in a pouch filled with an inert gas (e.g. argon). However, as most users don't have access to a sealing device, an alternative method would be to store the slides inside the 25-slide mailer boxes in a desiccator.

How can I make the deactivation solution for NEXTERION® Slide H?

There are two alternative deactivation solutions that could be used:
Solution 1: Solution 2:
100 mM boric acid
25 mM ethanolamine
0.01 % Tween 20
pH = 8.5

Preparation of 400 ml:
100 ml 400 mM boric acid pH 4.2
50 ml 200 mM ethanolamine
4 ml 1 % Tween 20
Add diH2O up to 400 ml
50 mM ethanolamine in PBST (0.01% Tween 20)
(see below)

Preparation of 1000 ml:
100 ml 10x PBS
10 ml 1% Tween 20
3 ml ethanolamine
Add diH2O up to 1000 ml

10x PBS
Preparation of 1000 ml:
80 g NaCl
15.795 g Na2HPO4
2.433 g Na2HPO4 x 2 H2O
2 g KCl
Dissolve in 500 ml diH2O
Add diH2O up to 1000 ml

NEXTERION® Slide H has a three dimensional coating. Are there any issues that I need to know about if I scan these slides with a (MDS Axon) GenePix® scanner that has a user-selectable focus position?

The coating of NEXTERION® Slide H stabilizes at around 50 nm when fully hydrated.
We recommend for optimal detection, that the "Focus position" (or offset) in the "Hardware settings" is set to +45 um This applies to GenePix® models 4000B, 4200A, 4300A, and 4400A.

I want to print on to NEXTERION® Slide H - Can you recommend a method of preparing the print buffer (150mM sodium phosphate buffer at pH 8.5)?

Prepare the following two solutions:

Solution #1 (e.g. 10 ml): 150 mM NaH2PO4 in DDH2O, pH should be approximately 4 (acidic component)
Solution #2 (e.g. 100 ml): 150 mM Na2HPO4 in DDH2O, pH should be approximately 9 (alkaline component)

The print buffer should be prepared immediately before printing:
Titrate solution 2 with solution 1 until the pH of solution 2 is 8.5. You will only need a few ml of solution 1. Afterwards you can add Tween® 20 and Sarkosyl e.g. from a 10% stock solution, to create 0.001 % Tween® 20 in the final solution.

Solution 1 and 2 can be stored for 6 months if kept sterile.

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