Slides of microcell hybrids and ZFN-transfected clones were stained with quinacrine mustard and Hoechst 33258 to enumerate chromosomes. Images were captured using an AxioImagerZ2 fluorescence microscope (Carl Zeiss GmbH, Jena, Germany). FISH analyses were performed using fixed metaphases of microcell hybrids using digoxigenin-labelled (Roche, Basel, Switzerland) human Cot-1 DNA (Life Technologies) and biotin-labelled (Roche) pSTneo plasmid DNA essentially as described previously20 (link). Chromosomal DNA was counterstained with DAPI (Sigma-Aldrich). Images were captured using the NIS-Elements system (Nikon, Tokyo, Japan). mFISH analyses were performed in accordance with the manufacturer instructions (MetaSystems, Altlussheim, Germany). Human mFISH probes were purchased from MetaSystems GmbH. Metaphase images were captured digitally with a CoolCubeI CCD camera and the ISIS mFISH software program (MetaSystems).
Coolcubei ccd camera
Manufactured by MetaSystems
The CoolCubeI CCD camera is a high-performance imaging device designed for scientific and industrial applications. It features a charge-coupled device (CCD) sensor that captures images with high resolution and sensitivity. The camera is capable of low-noise operation and offers various adjustable parameters to optimize image quality. The CoolCubeI provides a robust and reliable solution for imaging applications that require precise data capture.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Merck Group (3 mentions)
Human mfish probes, by MetaSystems (2 mentions)
Isis mfish software program, by MetaSystems (2 mentions)
Axio imager z2, by Zeiss (2 mentions)
Digoxigenin labeled, by Roche (2 mentions)
Nis elements system, by Nikon (1 mentions)
Human cot 1 dna, by Thermo Fisher Scientific (1 mentions)
Ikaros software program, by MetaSystems (1 mentions)
Isis mfish software, by MetaSystems (1 mentions)
Ikaros software, by MetaSystems (1 mentions)
3 protocols using coolcubei ccd camera
1
Chromosome Enumeration and FISH Analysis of Microcell Hybrids
2
Fluorescence In Situ Hybridization of Microcell Hybrids
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FISH analyses were performed in fixed metaphases of microcell hybrids using digoxigenin-labeled (Roche, Basel, Switzerland) human Cot-1 DNA (Life Technologies, Carlsbad, CA, USA) and biotin-labeled EGFP loxP or VEGF loxP vector. The preparation of chromosomes, probe, hybridization, washing, and signal detection were performed according to our previous report22 (link). Briefly, chromosomal DNA was counterstained with DAPI (Sigma). Images were captured using an AxioImagerZ2 fluorescence microscope (Carl Zeiss GmbH, Jena, Germany) and analyzed with the Ikaros software program (MetaSystems, Altlussheim, Germany). Multi-color FISH (mFISH) analyses were performed in accordance with the manufacturer’s instructions (MetaSystems)42 (link). CHO mFISH probes were purchased from MetaSystems GmbH. Metaphase images were captured digitally with a CoolCubeI CCD camera and the ISIS mFISH software program (MetaSystems).
3
Chromosome Analysis of Microcell Hybrids
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Microcell hybrids on slides were stained with quinacrine mustard and Hoechst 33258 to enumerate chromosomes. Images were captured under an AxioImagerZ2 fluorescence microscope (Carl Zeiss, Jena, Germany) and analyzed with Ikaros software (MetaSystems, Altlussheim, Germany). mFISH analyses were performed in accordance with the manufacturer instructions (MetaSystems). Human mFISH probes were purchased from MetaSystems. Metaphase images were captured digitally with a CoolCubeI CCD camera and ISIS mFISH software (MetaSystems). FISH analyses were performed on fixed metaphases of microcell hybrids using digoxigenin-labeled (Roche, Basel, Switzerland) alphoid DNA probe p11-443 (link) and biotin-labeled BAC DNA (RP11-954B16, located in the dystrophin genomic region) to detect DYS-HAC, and digoxigenin-labeled p11-4 and a biotin-labeled plasmid containing KO1 (ΦC31 attB-NeoR-pUbc-KO1) to confirm gene loading on the HAC.44 (link) Chromosomal DNA was counterstained with 4’,6-diamidino-2-phenylindole (DAPI) (Sigma-Aldrich). Images were captured under the AxioImagerZ2 fluorescence microscope.
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