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Cytovision version 7

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Cytovision version 7.2 is a software platform for digital microscopy and imaging. It provides a comprehensive suite of tools for acquiring, analyzing, and managing digital images from a variety of microscopy techniques.

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Lab products found in correlation

Dfc365 fs ccd camera, by Leica (2 mentions) Alexa fluor 488 5 dutp, by Thermo Fisher Scientific (2 mentions) Af594, by Thermo Fisher Scientific (2 mentions) Na plan apo objective, by Leica (2 mentions) Gsl 120 automated slide scanner, by Leica (2 mentions) Leitz dmrbe, by Leica (1 mentions) Texas red 5 dutp, by PerkinElmer (1 mentions) Bx53 fluorescence microscope, by Leica (1 mentions) Cellulase r 10, by PhytoTechnology Laboratories (1 mentions) Pectolyase y 23, by Duchefa Biochemie (1 mentions)

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Cytovision (28 citations)

8 protocols using cytovision version 7

1

Cytogenetic Analysis via C-banding and AgNOR

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C-banding and AgNOR-staining were performed according to previously published protocols [15 (link)][16 (link)]. Images were captured and processed using the software CytoVision version 7.2 (Leica Microsystems, Gateshead, UK) and JAI CV-M4+CL 1.4 megapixel progressive scan monochrome camera (JAI, Glostrup Copenhagen, DK) mounted on a Leica Leitz DMRBE (Leica Microsystems) fluorescence microscope.
Trifonov V.A., Paoletti A., Caputo Barucchi V., Kalinina T., O’Brien P.C., Ferguson-Smith M.A, & Giovannotti M. (2015). Comparative Chromosome Painting and NOR Distribution Suggest a Complex Hybrid Origin of Triploid Lepidodactylus lugubris (Gekkonidae). PLoS ONE, 10(7), e0132380.
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2

Fluorescent In Situ Hybridization of Ginseng Chromosomes

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Root mitotic chromosome spreads were prepared from stratified seeds provided by the Korea P. ginseng Corporation Natural Resources Research Institute (Daejeon, Korea), according to the methods described by Waminal et al [24] (link). Pg167TR and B. oleracea 5S rDNA amplicons were labeled with Texas Red-5-dUTP (NEL417001EA; Perkin Elmer, Waltham, MA, USA), and a 9-kb fragment of 45S rDNA (18S–5.8S–25S) [30] (link) was labeled with Alexa Fluor 488-5-dUTP (C11397; Invitrogen, Waltham, MA, USA) through direct nick-translation labeling. FISH procedures were performed as described previously [24] (link). Briefly, slides were immediately used for FISH after fixation with 4% paraformaldehyde, without pepsin and RNase pretreatment. Images were captured with an Olympus BX53 fluorescence microscope equipped with a Leica DFC365 FS CCD camera, and processed using Cytovision version 7.2 (Leica Microsystems, Wetzlar, Germany). Further image enhancements and creation of the idiogram were performed in Adobe Photoshop CC.
Waminal N.E., Choi H.I., Kim N.H., Jang W., Lee J., Park J.Y., Kim H.H, & Yang T.J. (2016). A refined Panax ginseng karyotype based on an ultra-high copy 167-bp tandem repeat and ribosomal DNAs. Journal of Ginseng Research, 41(4), 469-476.
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Chromosome Spread Preparation and FISH Analysis of Panax Species

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Preparation of P. notoginseng, P. ginseng, and P. quinquefolius chromosome spreads and FISH procedures were performed according to dual-color FISH analysis protocols49 (link). Briefly, root tips were treated with 2 mM 8-hyroxyquinoline, fixed with Carnoy’s solution, and were enzymatically digested with pectolytic enzyme solution (2% Cellulase R-10 (C224, Phytotechnology Laboratories) and 1% Pectolyase Y-23 (P8004.0001, Duchefa)) in 100 mM citrate buffer) for 1 h. Root tips were then squashed onto slides pre-cleaned with 70% ethanol. Air-dried slides were fixed in 2% formaldehyde for 5 min and dehydrated with a series of ethanol treatments (70%, 90%, and 100%)50 (link). PgDel1 and PgDel2 probes were obtained by PCR amplification using P. ginseng genomic DNA and primers detailed in our previous study27 (link). PgDel1 was labeled with Cy5-dUTP (Jena Bioscence), whereas PgDel2 was labeled with Diethyl amino coumarin-5-dUTP (NEL455001EA, Perkin Elmer) or Alexa Fluor 488-5-dUTP (C11397, Life Technologies). Images were captured using an Olympus BX53 epifluorescence microscope equipped with a Leica DFC365 FS CCD camera, and processed using Cytovision version 7.2 (Leica Microsystems, Germany). Further image enhancements were performed using Adobe Photoshop CS6.
Lee J., Waminal N.E., Choi H.I., Perumal S., Lee S.C., Nguyen V.B., Jang W., Kim N.H., Gao L.Z, & Yang T.J. (2017). Rapid amplification of four retrotransposon families promoted speciation and genome size expansion in the genus Panax. Scientific Reports, 7, 9045.
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Fluorescence In Situ Hybridization for chr2 Abnormalities

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For detection of chr2 abnormalities, two BAC clones were used as probes, one located distal to the BCL11A locus (2p21) as the telomeric marker and a clone from 2q11.2 as the centromeric marker. The telomeric BAC DNA (hg19 chr2:47612794 – 47782780) was labeled with a red-dUTP (AF594, Molecular Probes) by nick translation and the centromeric BAC DNA (hg19 chr2:99969552 – 100200667) was labeled with a green-dUTP (AF488, Molecular Probes). Both labeled probes were combined with sheared human DNA and hybridized in a solution containing 50 % formamide, 10 % dextran sulfate, and 2X SSC. The cells were then stained with 4,6-diamidino-2-phenylindole (DAPI) and imaged using a Nikon Eclipse 80i with a 100x/1.40 NA Plan Apo objective and Cytovision version 7.7 (Leica Biosystems).
Leibowitz M.L., Papathanasiou S., Doerfler P.A., Blaine L.J., Sun L., Yao Y., Zhang C.Z., Weiss M.J, & Pellman D. (2021). Chromothripsis as an on-target consequence of CRISPR-Cas9 genome editing. Nature genetics, 53(6), 895-905.
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5

Fluorescence In Situ Hybridization for chr2 Abnormalities

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For detection of chr2 abnormalities, two BAC clones were used as probes, one located distal to the BCL11A locus (2p21) as the telomeric marker and a clone from 2q11.2 as the centromeric marker. The telomeric BAC DNA (hg19 chr2:47612794 – 47782780) was labeled with a red-dUTP (AF594, Molecular Probes) by nick translation and the centromeric BAC DNA (hg19 chr2:99969552 – 100200667) was labeled with a green-dUTP (AF488, Molecular Probes). Both labeled probes were combined with sheared human DNA and hybridized in a solution containing 50 % formamide, 10 % dextran sulfate, and 2X SSC. The cells were then stained with 4,6-diamidino-2-phenylindole (DAPI) and imaged using a Nikon Eclipse 80i with a 100x/1.40 NA Plan Apo objective and Cytovision version 7.7 (Leica Biosystems).
Leibowitz M.L., Papathanasiou S., Doerfler P.A., Blaine L.J., Sun L., Yao Y., Zhang C.Z., Weiss M.J, & Pellman D. (2021). Chromothripsis as an on-target consequence of CRISPR-Cas9 genome editing. Nature genetics, 53(6), 895-905.
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6

Metaphase Chromosome and Interphase FISH Analysis

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Metaphases and nuclei were isolated in hypotonic buffer (0.25% KCl, 1% Na3C6H5O7), fixed with methanol:acetic acid (3:1), and dropped onto slides. Metaphase chromosomes were visualized with DAPI staining. Interphase FISH used chromosome enumeration probes labeled in Spectrum orange/green/aqua (Abbot Molecular) prepared in CEP hybridization buffer (Abbot Molecular). Slides were scanned on the GSL-120 automated slide scanner and analyzed using Cytovision version 7.3.1 (Leica Biosystems).
Martin C.A., Murray J.E., Carroll P., Leitch A., Mackenzie K.J., Halachev M., Fetit A.E., Keith C., Bicknell L.S., Fluteau A., Gautier P., Hall E.A., Joss S., Soares G., Silva J., Bober M.B., Duker A., Wise C.A., Quigley A.J., Phadke S.R., Wood A.J., Vagnarelli P, & Jackson A.P. (2016). Mutations in genes encoding condensin complex proteins cause microcephaly through decatenation failure at mitosis. Genes & Development, 30(19), 2158-2172.
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7

Chromosome Enumeration by FISH Analysis

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Nuclei cell suspensions were made by standard methods from patient fibroblasts incubated in hypotonic buffer (0.25% KCl) prior to fixation with 3:1 v/v methanol:acetic acid. FISH was performed using chromosome 3, 4, 7, 10, 17 and 18 centromere chromosome enumeration (CEP) probes (Abbot Molecular) labelled in Spectrum orange/green/aqua in CEP hybridization buffer. Slides were scanned on the GSL-120 Automated Slide Scanner at 60x magnification and analysed using Cytovision version 7.3.1 (Leica Biosystems).
Martin C.A., Ahmad I., Klingseisen A., Hussain M.S., Bicknell L.S., Leitch A., Nürnberg G., Toliat M.R., Murray J.E., Hunt D., Khan F., Ali Z., Tinschert S., Ding J., Keith C., Harley M.E., Heyn P., Müller R., Hoffmann I., Cormier-Daire V., Dollfus H., Dupuis L., Bashamboo A., McElreavey K., Kariminejad A., Mendoza-Londono R., Moore A.T., Saggar A., Schlechter C., Weleber R., Thiele H., Altmüller J., Höhne W., Hurles M.E., Noegel A.A., Baig S.M., Nürnberg P, & Jackson A.P. (2014). Mutations in PLK4, encoding a master regulator of centriole biogenesis, cause microcephaly, growth failure and retinopathy. Nature genetics, 46(12), 1283-1292.
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8

Chromosome Enumeration by FISH Analysis

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Nuclei cell suspensions were made by standard methods from patient fibroblasts incubated in hypotonic buffer (0.25% KCl) prior to fixation with 3:1 v/v methanol:acetic acid. FISH was performed using chromosome 3, 4, 7, 10, 17 and 18 centromere chromosome enumeration (CEP) probes (Abbot Molecular) labelled in Spectrum orange/green/aqua in CEP hybridization buffer. Slides were scanned on the GSL-120 Automated Slide Scanner at 60x magnification and analysed using Cytovision version 7.3.1 (Leica Biosystems).
Martin C.A., Ahmad I., Klingseisen A., Hussain M.S., Bicknell L.S., Leitch A., Nürnberg G., Toliat M.R., Murray J.E., Hunt D., Khan F., Ali Z., Tinschert S., Ding J., Keith C., Harley M.E., Heyn P., Müller R., Hoffmann I., Cormier-Daire V., Dollfus H., Dupuis L., Bashamboo A., McElreavey K., Kariminejad A., Mendoza-Londono R., Moore A.T., Saggar A., Schlechter C., Weleber R., Thiele H., Altmüller J., Höhne W., Hurles M.E., Noegel A.A., Baig S.M., Nürnberg P, & Jackson A.P. (2014). Mutations in PLK4, encoding a master regulator of centriole biogenesis, cause microcephaly, growth failure and retinopathy. Nature genetics, 46(12), 1283-1292.
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