ZytoLight^® SPEC FGFR3/4p11 Dual Color Probe

The ZytoLight^® SPEC FGFR3/4p11 Dual Color Probe (PL41) is a direct-labeled FISH probe designed for the qualitative detection of amplifications involving the human FGFR3 gene and the detection of specific sequences on chromosome 4p11 in formalin-fixed, paraffin-embedded (FFPE) specimens. It is intended to be used in combination with the ZytoLight FISH-Tissue Implementation Kit.

Principle:

The fluorescence in situ hybridization (FISH) technique detects and visualizes specific nucleic acid sequences using fluorescently-labeled DNA fragments (FISH probes) that co-denature and hybridize with complementary target DNA strands. After stringent washing to remove unbound fragments, the hybridized probes are visualized with a fluorescence microscope equipped with specific excitation and emission filters for the fluorochromes.In this method, direct FISH probes co-denature with target DNA in formalin-fixed, paraffin-embedded (FFPE) sections and hybridize overnight at 37°C after initial denaturation at 75°C. DAPI counterstaining follows, and the resulting green (FGFR3) and orange (4p11) signals are visualized via specific filters. The ZytoLight^® system eliminates the need for antibody detection and identifies amplifications by enumerating the signals.

Components:

ZytoLight^® SPEC FGFR3/4p11 Dual Color Probe comprises of:

• ZyGreen labeled polynucleotides (excitation 503 nm/emission 528 nm), targeting sequences in the FGFR3 gene region (4p16.3).
• ZyOrange labeled polynucleotides (excitation 547 nm/emission 572 nm), targeting sequences in the 4p11 locus.
• Formamide-based hybridization buffer.

The probe is designed for qualitative detection of sequences in the FGFR3 gene region and chromosome 4p11 specific sequences in formalin-fixed, paraffin-embedded (FFPE) tissue specimens by FISH. It is used in combination with the ZytoLight^® FISH-Tissue Implementation Kit for specimen pretreatment, hybridization, and post-hybridization processing.

For the qualitative detection of human FGFR3 gene amplifications and chromosome 4p11 specific sequences by fluorescence in situ hybridization (FISH)

In a normal interphase nucleus, the expected results are two green signals (for FGFR3) and two orange signals (for the 4p11 locus). In cells with FGFR3 amplifications, there may be an increased number of green signals or clusters observed. Any unexpected signal patterns should be further investigated, as variations may indicate different genetic rearrangements.

ZytoLight is a registered trademark of ZytoVision GmbH in Germany