Conventional chromosome G-band karyotyping analysis of the cultures of peripheral blood and amniotic fluid blood cells was performed as previously described [12 (link)]. Leica CytoVision (Leica, USA) was used to capture images of mitotic metaphase chromosomes. The karyotyping results were identified and described upon agreement of the two examiners, with reference to the International System for Human Cytogenetic Nomenclature (ISCN 2016) [13 ].
Cytovision
Manufactured by Leica
Sourced in United States, Germany, United Kingdom
Cytovision is a digital imaging and analysis system designed for cytogenetic applications. It provides high-quality digital imaging and automated analysis capabilities for metaphase chromosome preparations and other cytogenetic samples.
Automatically generated - may contain errors
Lab products found in correlation
Dxz1 dyz3, by Abbott (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (2 mentions)
Thermobrite, by Abbott (2 mentions)
Dm600b microscope, by Leica (1 mentions)
Axio imager, by Zeiss (1 mentions)
Giemsa solution, by Merck Group (1 mentions)
Agilent scanner, by Agilent Technologies (1 mentions)
Karyoarray 3, by Agilent Technologies (1 mentions)
Colcemid, by Irvine Scientific (1 mentions)
Dm5500 system, by Leica (1 mentions)
Cas block, by Thermo Fisher Scientific (1 mentions)
Giemsa stain solution, by Solarbio (1 mentions)
Colcemide, by Thermo Fisher Scientific (1 mentions)
Cell recovery solution, by Corning (1 mentions)
Dxz1 alpha satellite spectrumorange, by Abbott (1 mentions)
Dyz1 satellite 3 spectrumgreen, by Abbott (1 mentions)
Karyostudio software, by Illumina (1 mentions)
Karyomax colcemid solution, by Thermo Fisher Scientific (1 mentions)
Spectrum green dutp, by Abbott (1 mentions)
Spectrum orange dutp, by Abbott (1 mentions)
28 protocols using cytovision
1
Conventional Chromosome G-Band Karyotyping
2
Multicolor FISH Analysis of Hewga-CCS Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Metaphase chromosome spreads from Hewga-CCS cells were prepared according to standard procedures. Hewga-CCS cells were treated with 20 μg/ml of colcemide overnight and harvested. After treatment of 0.075 M KCl for 20 min at 37°C, cells were fixed 3 times with methanol and acetic acid (3:1) and fixed cells were spread on slides. Multicolor fluorescence in situ hybridization (M-FISH) was performed using commercially available M-FISH kits (MetaSystems, Altlussheim, Baden-Württemberg, Germany) according to the manufacturer’s protocol. Briefly metaphase spreads were hardened 70°C for 2 h. After applying M-FISH probes on the metaphase spreads, co-denaturation of target DNA with probe DNA was performed at 70°C for 5 min, followed by 72 h incubation at 37°C to allow hybridization of the probes. The slides were then washed twice with 50% formamide/2 × standard saline citrate (SSC) solution for 20 min at 37°C, 2 × SSC for 10 min at room temperature and 1 × SSC for 10 min. The slides were then counterstained with 4′,6-diamidino-2-phenylindole (DAPI) and mounted. Separate fluorochrome images were captured using a Leica DC 350FX cooled CCD camera (Leica Microsystems, Wetzlar, Hesse, Germany) mounted on a Leica DM600 B microscope using Leica DM600 B software. The images were analyzed using Leica CytoVision (Leica). The chromosomal analyses were examined at passage 110 and 111.
3
Conventional Chromosome G-Banding Karyotyping
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Conventional chromosome G-band karyotyping analysis of the cultures of peripheral blood and amniotic uid blood cells was performed as previously described [12] . Leica CytoVision (Leica, USA) was used to capture images of mitotic metaphase chromosomes. The karyotyping results were identi ed and described upon agreement of the two examiners, with reference to the International System for Human Cytogenetic Nomenclature (ISCN 2016) [13] .
4
MAML2 Gene Rearrangement Assessment
5
FISH Assay for TERC Amplification in NSCLC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
FISH assay was carried out on 4 μm (+ 1 μm) thick sections from formalin-fixed, paraffin-embedded tissue blocks from surgically resected tumor specimens of Non Small Cell Lung Cancer patients. The color TERC FISH probe was prepared with LSI TERC Spectrum Gold reagent (Abbott Molecular, Abbott Park, Illinois, U.S.A) according to the protocol previously described [33 (link)]. Analysis was performed on fluorescence microscope (Zeiss Axio Imager, Carl Zeiss S.p.A., Milan, Italy). For documentation, images were captured using a charge-coupled device camera (CoolSnap, Photometrics, Tucson, AR, USA) and merged using dedicated software (CytoVision, Leica Microsystems s.r.l., Milan, Italy). The scoring was carried out in 100 non-overlapping tumor cell nuclei per patient from four representative tumor areas. According to the Colorado criteria for epidermal growth factor receptor (EGFR) [33 (link)], the gene copy number for TERC was classified as increased (FISH-positive) when displaying gene amplification [>10% of tumor cells with >15 copies of the signals or gene clusters (>4 gene copies per cluster) or innumerable tight gene clusters] and high polysomy (≥40% of cells displaying ≥4 copies of the specific gene signal).
6
Mosaic 46,XY/47,XXY Karyotype and FISH Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Low-grade 46,XY/47,XXY mosaicism was present in two study patients (fraternal twins) and was confirmed by karyotype and FISH analysis at our institution. Standard protocols were used for karyotype analysis; 20 metaphase cells from Twin 1 and 50 metaphase cells from Twin 2 were evaluated. FISH was performed using spectrum orange labeled CEP X and spectrum green labeled LSI Yq12 probes (Abbot Molecular Inc, Des Plaines, IL, USA). Following overnight cohybridization, cells were washed, counterstained with diamidino-2-phenylindole and imaged using Leica Microsystems Cytovision (Buffalo Grove, IL, USA). Two hundred interphase nuclei from Twin 1 and 500 interphase nuclei plus 50 metaphase cells from Twin 2 were evaluated. The degree of mosaicism in the two 47,XXY/46,XY study patients was compared with karyotype and FISH results.
7
FISH Analysis of MYC and MYCN Amplification in FFPE Tumors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Unstained microtome sections of the FFPE tumour samples were analysed for C-MYC and MYCN amplification by FISH. The procedure was carried out at Camelia Botnar Laboratory, Great Ormond Street Hospital. FFPE sections of 4 μm thickness on glass slides were dried and de-waxed by heating to 60 °C in a dry oven, then washed in Xylene. The xylene was removed by passing the slides through a series of ethanol washes. The slides were then placed in a saline sodium citrate (SSC 2X) solution at 70 °C for 1 h. After washing in distilled water the slides were then digested using a 4 mg Pepsin solution in 0.2 N hydrochloric acid solution at 37 °C for 20 min. The slides were then washed in distilled water and dehydrated by passing through a series of alcohol washes. FISH probe sets for MYC/CEP8 and MYC/CEP8/IGH (Abbott, USA) and MYCN/AFF3 (Leica, Germany) were prepared and hybridised according to the manufacturers’ instructions. Cell images were captured using Olympus BX61 (Olympus, Japan) and Zeiss Axioskop Imager 1 (Zeiss, Germany) microscopes; image analysis was performed using Cytovision (Leica, UK), Isis (Metasystems, Germany) and SmartCapture (Digital Scientific, UK) software. For analysis, a target number of 100 representative informative cells were examined from each hybridisation, with 50 cells being scored independently by two analysts.
8
Cytogenetic Evaluation of hBM-MSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cytogenetic evaluation by G-banding method was conducted on hBM-MSCs at passages 3-4 and 6-7. Colchicine was added into each culture at a final concentration of 0.2 μg/ml for 4 hours at 37°C. MSCs were harvested using trypsin and resuspended in a hypotonic 0.075 mM KCl solution for 30 min at 37°C. After centrifugation the cells were fixed with methanol:acetic acid (3:1) solution. After dropping the cell suspension onto microscope slides, these were trypsinized and stained with Giemsa solution (Sigma-Aldrich, Germany). Slides were scanned, metaphases we captured and analyzed with Leica CytoVision® (USA) platform. 7 to 17 metaphase spreads were analyzed for chromosome number and structure abnormalities at each established passage. Karyotypes were described following the recommendations of the International System for Human Cytogenetic Nomenclature 2013 [97 ].
9
Karyotype and Array CGH Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The classical cytogenetics G-banding technique was performed after the patient had given birth to do a blood karyotype analysis. The karyotype analysis was performed on cells in metaphase using a microscope and the CytoVision (Leica) karyotype software system. After extracting DNA from the patient’s peripheral blood cells, 1 μg was used for array comparative genomic hybridization (a-CGH) utilizing KaryoArray 3.0 (8 × 60K; Agilent Technologies, Santa Clara, CA, United States) and was marked by fluorescence to compare the patient’s DNA with the control sample. DNA hybridization was done with a human genomic microarray of 860 K oligonucleotides using commercially available Agilent-based arrays, which were analyzed using an Agilent scanner with Feature Extraction 9.1 software. The aberration detection method 2 algorithm was used to determine any statistically significant aberrations. This was defined as the minimum number of oligonucleotides to consider an alteration. Subsequently, the medium resolution of the array was one oligonucleotide per 9 Kb in the regions of maximum interest, such as microdeletions and microduplications, centromeres, and telomeres. The algorithm in CGH Analytics version 3.5 software (Agilent Technologies) was used (Blanco-Kelly et al., 2017 (link)), as is described previously in the literature (Candelo et al., 2019 (link)).
10
Chromosome Preparation and Imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were treated with 20 ng/mL colcemid (Irvine Scientific, USA) for 5 h, and then harvested. After treatment with 0.075 M KCl for 25 min at 37°C, cells were fixed by exposure to MeOH:acetic acid (3:1) (repeated three times), and the fixed cells were spread on slides. Chromosome images were captured using a Cytovision (Leica, Wetzlar, Germany).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!