structured illumination microscopy (sim) imaging Search Results


structured illumination microscopy (sim) imaging  (SAS institute)
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SAS institute structured illumination microscopy (sim) imaging
Centriole overduplication in breast cancer cells that have CA. (A) Nonamplified and amplified centrioles have a full complement of representative centriolar proteins. SAS-6 (left, grayscale), CPAP (middle, grayscale) and CEP135 (right, grayscale) are labeled relative to CEP192 (red) and centrin (green) and imaged using <t>structured</t> <t>illumination</t> <t>microscopy</t> <t>(SIM).</t> (B) New centriole assembly at nonamplified and amplified centrioles. Left panel, centrioles labeled for SAS-6 (grayscale), CEP192 (red), and centrin (green). Right panel, schematic of G1-, early and late S phase, and amplified centrioles. Arrows denote multiple new, SAS-6–positive procentrioles. (C) Percentage of the total cell population that have more than two new (SAS-6–positive) daughter centrioles. (D) Number of new centrioles (SAS-6 positive) assembled in cells that have amplified centrosomes. (E) Multiple and single daughter centrioles form from mother centrioles. Top panel, schematic of multiple and single daughter centrioles forming from mother centrioles. Bottom panel, representative images of centrioles labeled for SAS-6 (grayscale), CEP192 (red), and centrin (green) in S-phase MDA-231 cells that have overduplicated centrioles. Arrows denote multiple daughter centriole assembly events at a single mother centriole. (F) Relative frequency of duplication of multiple daughter centrioles compared with a single daughter centriole from a single mother centriole. (C, D) Statistical tests compare to MCF10A cells. Mean ± SEM. Fisher’s exact test and Mann–Whitney U test. ***, p < 0.005 and ****, p < 0.0005. Scale bars, 1 μm.
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Centriole overduplication in breast cancer cells that have CA. (A) Nonamplified and amplified centrioles have a full complement of representative centriolar proteins. SAS-6 (left, grayscale), CPAP (middle, grayscale) and CEP135 (right, grayscale) are labeled relative to CEP192 (red) and centrin (green) and imaged using structured illumination microscopy (SIM). (B) New centriole assembly at nonamplified and amplified centrioles. Left panel, centrioles labeled for SAS-6 (grayscale), CEP192 (red), and centrin (green). Right panel, schematic of G1-, early and late S phase, and amplified centrioles. Arrows denote multiple new, SAS-6–positive procentrioles. (C) Percentage of the total cell population that have more than two new (SAS-6–positive) daughter centrioles. (D) Number of new centrioles (SAS-6 positive) assembled in cells that have amplified centrosomes. (E) Multiple and single daughter centrioles form from mother centrioles. Top panel, schematic of multiple and single daughter centrioles forming from mother centrioles. Bottom panel, representative images of centrioles labeled for SAS-6 (grayscale), CEP192 (red), and centrin (green) in S-phase MDA-231 cells that have overduplicated centrioles. Arrows denote multiple daughter centriole assembly events at a single mother centriole. (F) Relative frequency of duplication of multiple daughter centrioles compared with a single daughter centriole from a single mother centriole. (C, D) Statistical tests compare to MCF10A cells. Mean ± SEM. Fisher’s exact test and Mann–Whitney U test. ***, p < 0.005 and ****, p < 0.0005. Scale bars, 1 μm.

Journal: Molecular Biology of the Cell

Article Title: CEP135 isoform dysregulation promotes centrosome amplification in breast cancer cells

doi: 10.1091/mbc.E18-10-0674

Figure Lengend Snippet: Centriole overduplication in breast cancer cells that have CA. (A) Nonamplified and amplified centrioles have a full complement of representative centriolar proteins. SAS-6 (left, grayscale), CPAP (middle, grayscale) and CEP135 (right, grayscale) are labeled relative to CEP192 (red) and centrin (green) and imaged using structured illumination microscopy (SIM). (B) New centriole assembly at nonamplified and amplified centrioles. Left panel, centrioles labeled for SAS-6 (grayscale), CEP192 (red), and centrin (green). Right panel, schematic of G1-, early and late S phase, and amplified centrioles. Arrows denote multiple new, SAS-6–positive procentrioles. (C) Percentage of the total cell population that have more than two new (SAS-6–positive) daughter centrioles. (D) Number of new centrioles (SAS-6 positive) assembled in cells that have amplified centrosomes. (E) Multiple and single daughter centrioles form from mother centrioles. Top panel, schematic of multiple and single daughter centrioles forming from mother centrioles. Bottom panel, representative images of centrioles labeled for SAS-6 (grayscale), CEP192 (red), and centrin (green) in S-phase MDA-231 cells that have overduplicated centrioles. Arrows denote multiple daughter centriole assembly events at a single mother centriole. (F) Relative frequency of duplication of multiple daughter centrioles compared with a single daughter centriole from a single mother centriole. (C, D) Statistical tests compare to MCF10A cells. Mean ± SEM. Fisher’s exact test and Mann–Whitney U test. ***, p < 0.005 and ****, p < 0.0005. Scale bars, 1 μm.

Article Snippet: The number of SAS-6 foci at each mother centriole was quantified using structured illumination microscopy (SIM) imaging of MDA-231 cells that exhibit greater than two procentrioles (SAS-6 positive) per cell.

Techniques: Amplification, Labeling, Microscopy, MANN-WHITNEY