Journal: Nucleic Acids Research

Article Title: A large field CCD system for quantitative imaging of microarrays

doi: 10.1093/nar/gkl160

Figure Lengend Snippet: Detection sensitivity. ( a ) Glycerol-PBS solutions containing known concentrations of Cy3 and Cy5 were loaded into rectangular fused silica capillaries, placed on a chromium-coated microscope slide, surrounded by glycerol and a glass cover slip applied. This mounting closely matched the indices of refraction of capillaries (fused silica ∼1.459, glycerol ∼1.473) so that very little light scatter was generated at the capillary surfaces. Five capillaries were mounted in an image field, each containing a mixture of Cy3 and Cy5 at concentrations of 0, 1, 10, 100 and 1000 molecules/µm 2 , and Cy3 and Cy5 images acquired. Concentrations of stock solutions were determined using absorbance measurements and known extinction coefficients. ( b ) Fluorescence intensities Cy3 (red curve) and Cy5 (black curve) increased linearly with fluorochrome density. The inset shows the portion of the curves between 0 and 10 fluorochromes/µm 2 , indicting the ability to detect concentrations below 1 fluorochrome/µm 2 .

Article Snippet: Solutions containing various concentrations of Cy3 and Cy5 in the glycerol/10% PBS medium we use for slide mounting were loaded into rectangular fused silica capillaries of interior dimensions 20 μm × 200 μm (Vitrotubes #5002S-050; VitroCom, Mountain Lakes NJ), and the capillaries were mounted on a chromium array substrate, as shown in .

Techniques: Microscopy, Generated, Fluorescence