UniStar - a Universal In Situ Hybridization Assay designed for FISH and CISH on the Same Specimen
Chromogenic detection via bright field microscopy is the method of choice for many pathologists who do not have acess to a fluorescence microscope, or who simply do not see the need to perform FISH, in particular as recent developments have made colorimetric detection equivalent to FISH in terms of accuracy. In addition, by using a brightfield microscope, tissue morphology can be studied more easily, and signals can be archived for later analysis.
Upon mounting of the formalin-fixed paraffin-embedded tissue section onto a microscope slide, a conventional FISH assay is performed by pretreating the slide to enable efficient hybridization of a fluorescent DNA probe. Subsequently, the signals can be analyzed with a fluorescent microscope followed by conversion of the FISH signal into a chromogenic signal followed by imaging using a conventional bright field microscope (A). Alternatively, the hybridized probes can directly be analyzed with CISH (B).
The first UniStar assays introduced are designed for detecting Her2/Neu, EGFR, and the membrane bound receptor C-MET, respectively. Each of these kits include the corresponding DNA probes for copy number detection of the gene of interest, as well as a control probe for performing FISH in a dual-color assay. In addition, they include a specific detection module converting the signal of the critical probe into a chromogenic single-color signal via a colorimetric assay system.
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FISH
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UniStar CISH
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Breast cancer tissue hybridized with the Her-2Neu/ SE 7 POSEIDON fluorescent probe followed by conversion of red Her-2/Neu signal into a brownish chromogenic signal using the UniStar CISH Detection KIt.
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Glioblastoma specimen hybridized with the EGFR/ SE 7 POSEIDON fluorescent probe followed by conversion of the red EGFR signal into a colorimetric signal using the UniStar CISH Detection Kit.
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Colon carcinoma specimen showing a normal EGFR expression (non-amplified). The slide was hybridized with the EGFR/ SE 7 POSEIDON fluorescent probe followed by conversion into a colormetric signal using the UniStar CISH Detection Kit.
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Images Her-2 Neu kindly provided by Dr. M. Gosens and Dr. E. Moerland, Stichting PAMM, Eindhoven, The Netherlands.
Images EGFR kindly provided by Dr. K. Beiske, Oslo University Hospital, Norway.
UniStar - a Universal In Situ Hybridization Assay designed for FISH and CISH on the Same Specimen
Chromogenic detection via bright field microscopy is the method of choice for many pathologists who do not have acess to a fluorescence microscope, or who simply do not see the need to perform FISH, in particular as recent developments have made colorimetric detection equivalent to FISH in terms of accuracy. In addition, by using a brightfield microscope, tissue morphology can be studied more easily, and signals can be archived for later analysis.
Upon mounting of the formalin-fixed paraffin-embedded tissue section onto a microscope slide, a conventional FISH assay is performed by pretreating the slide to enable efficient hybridization of a fluorescent DNA probe. Subsequently, the signals can be analyzed with a fluorescent microscope followed by conversion of the FISH signal into a chromogenic signal followed by imaging using a conventional bright field microscope (A). Alternatively, the hybridized probes can directly be analyzed with CISH (B).
The first UniStar assays introduced are designed for detecting Her2/Neu, EGFR, and the membrane bound receptor C-MET, respectively. Each of these kits include the corresponding DNA probes for copy number detection of the gene of interest, as well as a control probe for performing FISH in a dual-color assay. In addition, they include a specific detection module converting the signal of the critical probe into a chromogenic single-color signal via a colorimetric assay system.
|
FISH
|
|
UniStar CISH
|
|
|
|
|
Breast cancer tissue hybridized with the Her-2Neu/ SE 7 POSEIDON fluorescent probe followed by conversion of red Her-2/Neu signal into a brownish chromogenic signal using the UniStar CISH Detection KIt.
|
|
|
|
|
Glioblastoma specimen hybridized with the EGFR/ SE 7 POSEIDON fluorescent probe followed by conversion of the red EGFR signal into a colorimetric signal using the UniStar CISH Detection Kit.
|
|
|
|
|
Colon carcinoma specimen showing a normal EGFR expression (non-amplified). The slide was hybridized with the EGFR/ SE 7 POSEIDON fluorescent probe followed by conversion into a colormetric signal using the UniStar CISH Detection Kit.
|
Images Her-2 Neu kindly provided by Dr. M. Gosens and Dr. E. Moerland, Stichting PAMM, Eindhoven, The Netherlands.
Images EGFR kindly provided by Dr. K. Beiske, Oslo University Hospital, Norway.