What can cause poor resolution when observing cells under a microscope?
Please review the following possible causes and solutions:
- The microscope could be dirty from fingerprints or oils. We recommend cleaning your microscope prior to use and always after oil immersion. When using oil immersion, it is important to use the specific brand of oil designed and recommended for that particular microscope.
- An incorrect type of coverslip or coverglass can cause poor resolution. We recommend Nunc Lab-Tek Chambered Coverglass (made from 1.0 borosilicate glass) and Nunc Lab-Tek II Chambered Coverglass (made from 1.5 borosilicate glass) for the best resolution. Plastic coverslips may affect resolution at high magnification due to refractive index differences between the glass and plastic.
- An incorrect thickness for the coverglass can cause poor resolution. Please consult your microscope's manual for the best information.
- The coverslipping procedure may affect resolution. A clear seal is required, free of debris and particles. When performing oil immersion, bubbles may result in poor resolution under high magnification.
How does cell culturing on Nunc Lab-Tek Chambered Coverglass compare with Nunc Lab-Tek II Chambered Coverglass?
Lab-Tek and Lab-Tek II chambered coverglass products can be plated with cells, incubated, and handled in the same way. The two products differ in the thickness of the glass and the chamber size. Lab-Tek coverglass uses Borosilicate coverglass No. 1 that is 0.13 - 0.16 mm thick and Lab-Tek II coverglass uses Borosilicate coverglass No. 1.5 thath is 0.16 - 0.19 mm thick. Lab-Tek II coverglass chambers hold 10 - 50% more media than Lab-Tek coverglass chambers, depending on the format. Studies have shown that Lab-Tek II chambered coverglass works well with fastidious cells such as neuronal, as well as poorly adherent cell lines.
What stains and fixatives are compatible with Lab-Tek chambered slides and coverglasses?
Lab-Tek products are compatible with all common fixatives, including paraformaldehyde. Acetone at 100% and some mixtures of acetone and alcohols are not compatible with the polystyrene upper well structure and will result in clouding or other effects of chemical attack. We encourage careful testing within planned operating conditions before working with high-value samples.
What factors must I consider when working with Lab-Tek and Lab-Tek II chambered coverglass products?
Lab-Tek chambered coverglasses are not accessories to Lab-Tek chamber slides, but another complete product for cell culture. The chambers are simply adhered to a borosilicate coverglass (to serve as the culture surface) rather than a soda lime glass or Permanox microscope slide. Most customers select this format for compatibility with confocal imaging systems that require a #1.0, approx. 120 micron (Lab-Tek) or #1.5, approx. 180 micron (Lab-Tek II) substrate. Because the coverglass is very thin and fragile, the chambered coverglasses (unlike chamber slides) cannot be disassembled.
What are the different options available in Lab-Tek II chamber slides?
Lab-Tek II chambered glass slide systems are available in our standard high-purity water washed culture treatment or our CC2 surface modification. The latter is a proprietary passive treatment that resembles poly-D-lysine, often improving culture performance when working with fastidious cells such as neuronal cultures.