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(A) Schematic shows a pair of homologous chromosomes localize on opposite sides of the nuclear hemisphere for both autosomal, and sex chromosomes in neonatal human vein endothelial cells (HUVECs) (Hua and Mikawa 2018). (B) Pairing is defined as the distance between two homologous chromosomes to be at 0 µm and the vector angle between the two homologous chromosomes at 0°. Different scenarios of not paired (a,b) vs paired (c) are shown. (C) Top view of optical sections of a HUVEC at metaphase stained for chromosomes 17 (cyan), γ-tubulin (blue), and counterstained for <t>DNA</t> (grey). Inset: 3D reconstruction of the optical sections. (D) A 3D overlay of chromosome 17 (cyan/grey) of multiple HUVECs at metaphase, where the homolog closest to the x-axis is colored (cyan), while its respective partner is in grey (n=12 cells). (E) Relative positions for each homolog of chromosome 17 (cyan/grey circles) when mapped to an axial coordinate system. (F) The distance and angular orientation of homologous chromosome 17 pairs with an average distance of 2.97 ± 1.07 µm SD and angular orientation of 92.5° ± 49.1°. Note: Each numbered cell corresponds to the same cell number in the relative position normalized to the DNA <t>counterstain</t> (E). (G-J) As in C-F, but of chromosome 19 (n=35 cells). The distance and angular orientation of chromosome 19 homologs with an average distance of 2.90 ± 1.74 µm SD and angular orientation of 114.2° ± 29.7°. (K-N) As in C-F, but of adult human aortic endothelial cells (HAECs) (n=4 individual patients, n=82 cells) with an average distance of 1.66 ± 1.39 µm SD and angular orientation of 101.0° ± 51.2°. (O-R) As in G-J, but of HAECs. The distance and angular orientation of chromosome 19 homologs (n=4 individual patients, n=38 cells) with an average distance of 2.02 ± 1.79 µm SD and angular orientation of 93.8° ± 55.8° SD. (S) Pairing frequency graph showing the pairing of chromosomes 17 (cyan) and 19 (magenta) of HUVECs and HAECs. Scale bar: 2 µm.
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(A) Schematic shows a pair of homologous chromosomes localize on opposite sides of the nuclear hemisphere for both autosomal, and sex chromosomes in neonatal human vein endothelial cells (HUVECs) (Hua and Mikawa 2018). (B) Pairing is defined as the distance between two homologous chromosomes to be at 0 µm and the vector angle between the two homologous chromosomes at 0°. Different scenarios of not paired (a,b) vs paired (c) are shown. (C) Top view of optical sections of a HUVEC at metaphase stained for chromosomes 17 (cyan), γ-tubulin (blue), and counterstained for <t>DNA</t> (grey). Inset: 3D reconstruction of the optical sections. (D) A 3D overlay of chromosome 17 (cyan/grey) of multiple HUVECs at metaphase, where the homolog closest to the x-axis is colored (cyan), while its respective partner is in grey (n=12 cells). (E) Relative positions for each homolog of chromosome 17 (cyan/grey circles) when mapped to an axial coordinate system. (F) The distance and angular orientation of homologous chromosome 17 pairs with an average distance of 2.97 ± 1.07 µm SD and angular orientation of 92.5° ± 49.1°. Note: Each numbered cell corresponds to the same cell number in the relative position normalized to the DNA <t>counterstain</t> (E). (G-J) As in C-F, but of chromosome 19 (n=35 cells). The distance and angular orientation of chromosome 19 homologs with an average distance of 2.90 ± 1.74 µm SD and angular orientation of 114.2° ± 29.7°. (K-N) As in C-F, but of adult human aortic endothelial cells (HAECs) (n=4 individual patients, n=82 cells) with an average distance of 1.66 ± 1.39 µm SD and angular orientation of 101.0° ± 51.2°. (O-R) As in G-J, but of HAECs. The distance and angular orientation of chromosome 19 homologs (n=4 individual patients, n=38 cells) with an average distance of 2.02 ± 1.79 µm SD and angular orientation of 93.8° ± 55.8° SD. (S) Pairing frequency graph showing the pairing of chromosomes 17 (cyan) and 19 (magenta) of HUVECs and HAECs. Scale bar: 2 µm.
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(A) Schematic shows a pair of homologous chromosomes localize on opposite sides of the nuclear hemisphere for both autosomal, and sex chromosomes in neonatal human vein endothelial cells (HUVECs) (Hua and Mikawa 2018). (B) Pairing is defined as the distance between two homologous chromosomes to be at 0 µm and the vector angle between the two homologous chromosomes at 0°. Different scenarios of not paired (a,b) vs paired (c) are shown. (C) Top view of optical sections of a HUVEC at metaphase stained for chromosomes 17 (cyan), γ-tubulin (blue), and counterstained for <t>DNA</t> (grey). Inset: 3D reconstruction of the optical sections. (D) A 3D overlay of chromosome 17 (cyan/grey) of multiple HUVECs at metaphase, where the homolog closest to the x-axis is colored (cyan), while its respective partner is in grey (n=12 cells). (E) Relative positions for each homolog of chromosome 17 (cyan/grey circles) when mapped to an axial coordinate system. (F) The distance and angular orientation of homologous chromosome 17 pairs with an average distance of 2.97 ± 1.07 µm SD and angular orientation of 92.5° ± 49.1°. Note: Each numbered cell corresponds to the same cell number in the relative position normalized to the DNA <t>counterstain</t> (E). (G-J) As in C-F, but of chromosome 19 (n=35 cells). The distance and angular orientation of chromosome 19 homologs with an average distance of 2.90 ± 1.74 µm SD and angular orientation of 114.2° ± 29.7°. (K-N) As in C-F, but of adult human aortic endothelial cells (HAECs) (n=4 individual patients, n=82 cells) with an average distance of 1.66 ± 1.39 µm SD and angular orientation of 101.0° ± 51.2°. (O-R) As in G-J, but of HAECs. The distance and angular orientation of chromosome 19 homologs (n=4 individual patients, n=38 cells) with an average distance of 2.02 ± 1.79 µm SD and angular orientation of 93.8° ± 55.8° SD. (S) Pairing frequency graph showing the pairing of chromosomes 17 (cyan) and 19 (magenta) of HUVECs and HAECs. Scale bar: 2 µm.
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(A) Schematic shows a pair of homologous chromosomes localize on opposite sides of the nuclear hemisphere for both autosomal, and sex chromosomes in neonatal human vein endothelial cells (HUVECs) (Hua and Mikawa 2018). (B) Pairing is defined as the distance between two homologous chromosomes to be at 0 µm and the vector angle between the two homologous chromosomes at 0°. Different scenarios of not paired (a,b) vs paired (c) are shown. (C) Top view of optical sections of a HUVEC at metaphase stained for chromosomes 17 (cyan), γ-tubulin (blue), and counterstained for <t>DNA</t> (grey). Inset: 3D reconstruction of the optical sections. (D) A 3D overlay of chromosome 17 (cyan/grey) of multiple HUVECs at metaphase, where the homolog closest to the x-axis is colored (cyan), while its respective partner is in grey (n=12 cells). (E) Relative positions for each homolog of chromosome 17 (cyan/grey circles) when mapped to an axial coordinate system. (F) The distance and angular orientation of homologous chromosome 17 pairs with an average distance of 2.97 ± 1.07 µm SD and angular orientation of 92.5° ± 49.1°. Note: Each numbered cell corresponds to the same cell number in the relative position normalized to the DNA <t>counterstain</t> (E). (G-J) As in C-F, but of chromosome 19 (n=35 cells). The distance and angular orientation of chromosome 19 homologs with an average distance of 2.90 ± 1.74 µm SD and angular orientation of 114.2° ± 29.7°. (K-N) As in C-F, but of adult human aortic endothelial cells (HAECs) (n=4 individual patients, n=82 cells) with an average distance of 1.66 ± 1.39 µm SD and angular orientation of 101.0° ± 51.2°. (O-R) As in G-J, but of HAECs. The distance and angular orientation of chromosome 19 homologs (n=4 individual patients, n=38 cells) with an average distance of 2.02 ± 1.79 µm SD and angular orientation of 93.8° ± 55.8° SD. (S) Pairing frequency graph showing the pairing of chromosomes 17 (cyan) and 19 (magenta) of HUVECs and HAECs. Scale bar: 2 µm.
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(A) Schematic shows a pair of homologous chromosomes localize on opposite sides of the nuclear hemisphere for both autosomal, and sex chromosomes in neonatal human vein endothelial cells (HUVECs) (Hua and Mikawa 2018). (B) Pairing is defined as the distance between two homologous chromosomes to be at 0 µm and the vector angle between the two homologous chromosomes at 0°. Different scenarios of not paired (a,b) vs paired (c) are shown. (C) Top view of optical sections of a HUVEC at metaphase stained for chromosomes 17 (cyan), γ-tubulin (blue), and counterstained for DNA (grey). Inset: 3D reconstruction of the optical sections. (D) A 3D overlay of chromosome 17 (cyan/grey) of multiple HUVECs at metaphase, where the homolog closest to the x-axis is colored (cyan), while its respective partner is in grey (n=12 cells). (E) Relative positions for each homolog of chromosome 17 (cyan/grey circles) when mapped to an axial coordinate system. (F) The distance and angular orientation of homologous chromosome 17 pairs with an average distance of 2.97 ± 1.07 µm SD and angular orientation of 92.5° ± 49.1°. Note: Each numbered cell corresponds to the same cell number in the relative position normalized to the DNA counterstain (E). (G-J) As in C-F, but of chromosome 19 (n=35 cells). The distance and angular orientation of chromosome 19 homologs with an average distance of 2.90 ± 1.74 µm SD and angular orientation of 114.2° ± 29.7°. (K-N) As in C-F, but of adult human aortic endothelial cells (HAECs) (n=4 individual patients, n=82 cells) with an average distance of 1.66 ± 1.39 µm SD and angular orientation of 101.0° ± 51.2°. (O-R) As in G-J, but of HAECs. The distance and angular orientation of chromosome 19 homologs (n=4 individual patients, n=38 cells) with an average distance of 2.02 ± 1.79 µm SD and angular orientation of 93.8° ± 55.8° SD. (S) Pairing frequency graph showing the pairing of chromosomes 17 (cyan) and 19 (magenta) of HUVECs and HAECs. Scale bar: 2 µm.

Journal: bioRxiv

Article Title: Higher frequency of homologous chromosome pairing in human adult aortic endothelial cells

doi: 10.1101/2025.03.15.643486

Figure Lengend Snippet: (A) Schematic shows a pair of homologous chromosomes localize on opposite sides of the nuclear hemisphere for both autosomal, and sex chromosomes in neonatal human vein endothelial cells (HUVECs) (Hua and Mikawa 2018). (B) Pairing is defined as the distance between two homologous chromosomes to be at 0 µm and the vector angle between the two homologous chromosomes at 0°. Different scenarios of not paired (a,b) vs paired (c) are shown. (C) Top view of optical sections of a HUVEC at metaphase stained for chromosomes 17 (cyan), γ-tubulin (blue), and counterstained for DNA (grey). Inset: 3D reconstruction of the optical sections. (D) A 3D overlay of chromosome 17 (cyan/grey) of multiple HUVECs at metaphase, where the homolog closest to the x-axis is colored (cyan), while its respective partner is in grey (n=12 cells). (E) Relative positions for each homolog of chromosome 17 (cyan/grey circles) when mapped to an axial coordinate system. (F) The distance and angular orientation of homologous chromosome 17 pairs with an average distance of 2.97 ± 1.07 µm SD and angular orientation of 92.5° ± 49.1°. Note: Each numbered cell corresponds to the same cell number in the relative position normalized to the DNA counterstain (E). (G-J) As in C-F, but of chromosome 19 (n=35 cells). The distance and angular orientation of chromosome 19 homologs with an average distance of 2.90 ± 1.74 µm SD and angular orientation of 114.2° ± 29.7°. (K-N) As in C-F, but of adult human aortic endothelial cells (HAECs) (n=4 individual patients, n=82 cells) with an average distance of 1.66 ± 1.39 µm SD and angular orientation of 101.0° ± 51.2°. (O-R) As in G-J, but of HAECs. The distance and angular orientation of chromosome 19 homologs (n=4 individual patients, n=38 cells) with an average distance of 2.02 ± 1.79 µm SD and angular orientation of 93.8° ± 55.8° SD. (S) Pairing frequency graph showing the pairing of chromosomes 17 (cyan) and 19 (magenta) of HUVECs and HAECs. Scale bar: 2 µm.

Article Snippet: Slides were then washed twice with 1x PBS, and processed with DNA counterstain DAPI (Invitrogen: D1306) for immunofluorescence.

Techniques: Plasmid Preparation, Staining