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Encryption Algorithm In Algorithms In Web Development}FISH-PALM: A FLUorescence IN Situ Hybridization-Phase-Contrast Microscopy System for Whole-Slide Imaging of Fixed, Formalin-Fixed, and Paraffin-Embedded Tissue Sections.
FISH-PALM is a new confocal fluorescence in situ hybridization (FISH) microscopy system that combines the ability to target, visualize, and quantitatively image multiple genes with the ability to simultaneously record and enhance the signal intensity of the counterstained phase-contrast images of fixed, formalin-fixed, and paraffin-embedded tissue sections. The combination of phase contrast microscopy (PCM) with FISH imaging of chromosome 3q amplification is illustrated using two breast cancer cell lines, T47D and SKBR3, showing that the addition of a fluorescent channel to a basic PCM system allows real-time detection of the hybridization patterns of one or more gene-specific probes without the need for time-consuming washing steps or post-hybridization incubation. In our hands, FISH-PALM provides superior detection limits (0.01-0.05 gene copies) compared with other FISH systems that use post-hybridization washing steps. In addition, the hybridization specificity is preserved, which is advantageous for distinguishing the loss of, or abnormal hybridization patterns of, some of the probes on specific chromosomes from the relative lack of hybridization to non-target chromosomes. Thus, FISH-PALM enables both the detection and visualization of multiple probes in a single image. We also describe the ability to extend the dynamic range of the FISH-PALM system by using multiple excitation lasers that individually and simultaneously illuminate the sections, thereby increasing the signal-to-noise ratio (SNR)
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