Filters digital microscopy

The application of in situ hybridization (ISH) has advanced from short lived, non-specific isotopic methods, to very specific, long lived, multiple color fluorescent-ISH probe assays (FISH). Improvements in the optics, filter technology, microscopes, cameras, and data handling by software, have allowed for a cost effective FISH setup to be within reach of most researchers. The application of mFISH (multiplex-FISH), coupled to the advances in digital imaging microscopy, have vastly improved the capabilities for non-isotopic detection and analysis of multiple nucleic acid sequences in chromosomes and genes (1). 

Specimens for microscopy were obtained from the bonded specimens and from the samples after the D 905 tests. For transverse sections, samples were water-soaked for 2-24 h prior to microtoming. The sections were analyzed using a scanning electron microscope (JEOL 840 after gold plating the samples) or a Leitz Orthoplan epi-fluorescence microscope with a 150-W mercury lamp light source, an A2 UV filter cube, and a Nikon DS-5M digital camera, or a Carl Zeiss Axioskop epi-fluorescence microscope with a 100-W mercury lamp, a UV filter set, and CCD camera. Fluorescence microscopy was used to examine the failure surface. 

A Nanoscope IIIA Multimode (Digital Instruments, Santa Barbara, CA) was used for the atomic force microscopy measurements. The AFM was operated in the force mode, with a scan rate of 1 Hz and a z-piezo total displacement of 500 nm. Both approach/extension and retraction force curves of the cantilever were recorded. The experiments were performed in liquid, using an open fluid cell and letting the system equilibrate for 30—60 min. All buffers were filtered through a 0.22 fj,m pore size filter. 

---