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Cell Structure Observations

Observations about microscopy videos for each of the 16 cell lines available in our Cell Catalog & 3D Cell Viewer.

Endoplasmic reticulum & nuclear envelope via Sec61β/Lamin B1

2/20/2019

 
Movie. High magnification Z-stack of a live hiPS cell colony expressing mEGFP-tagged Sec61 beta and mTagRFP-T-tagged lamin B1. Panels show individual channels for Sec61 beta (left) and lamin B1 (middle) and the overlay of the two (right). Cells were imaged in 3D on a spinning-disk confocal microscope. Movie starts at the bottom of the cells and ends at the top. Scale bar, 5µm.

Movie. Time-lapse in high magnification movie of a live hiPS cell colony expressing mEGFP-tagged Sec61 beta and mTagRFP-T-tagged lamin B1. Panels show individual channels for Sec61 beta (left) and lamin B1 (middle) and the overlay of the two (right). A single, mid-level plane was imaged on a spinning-disk confocal microscope every 5 min. Movie plays at 3000x real time. Scale bar, 5 µm.

Movie. Time-lapse in low magnification movie of a live hiPS cell colony expressing mEGFP-tagged Sec61 beta and mTagRFP-T-tagged lamin B1. Panels show individual channels for Sec61 beta (left) and lamin B1 (middle) and the overlay of the two (right). Cells were imaged in 3D on a spinning-disk confocal microscope every 5 min. A single mid-level z-section is shown. Movie plays at 1200x real time. Scale bar, 20 µm.

Observations
  • We previously tagged Sec61 beta and Lamin B1 with mEGFP in two separate cell lines (AICS-0010 and AICS-0013, respectively). The localization of mEGFP-tagged Sec61 beta and mTagRFP-T-tagged lamin B1 in our dual-edited line does not differ from the localization of mEGFP-tagged Sec61 beta and mEGFP-tagged lamin B1 in our previously validated lines.
  • In interphase, the two tagged proteins co-localize to the nuclear envelope, and only Sec61 beta is found in the ER sheets and tubules throughout the cytoplasm. During cell division, both proteins co-localize to a dim extended wavy morphology. At the end of division, Sec61 beta and lamin B1 localize around the re-forming nucleus. Lamin B1 appears as bright segments along the ER near the re-forming nucleus; the segments disappear by early G1. Just after division, lamin B1 and Sec61 beta co-localize within many invaginations within the nuclear envelope, which decrease in number with time. 

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  • About
      Institute
      1. News feed
      2. What we do
      3. Publications
      4. Allen Institute | allenInstitute.org
      5. Careers
      Site
      1. Home page
      2. Site updates
      3. Archived content
  • Allen Cell Collection
      Order cells & plasmids
      1. Cell Catalog
      2. Cell Catalog quickview
      3. Cell Shorts (documentaries on labs using our cells)
      4. Support forum
      Lab methods
      1. Instructional videos for success in the lab
      2. Standard operating procedures (written methods)
      3. Illustrated overviews
      About our hiPS cells
      1. hiPS Cell Structure Overview
      2. Visual Guide to Human Cells
      3. Cell structure observations
      4. Why endogenous tagging?
      5. Differentiation into cardiomyocytes
      6. Genomics
      7. Download cell data (images, genomics, features)
  • Data & Digital Tools
      Online image analysis
      1. Cell Feature Explorer (plotting & 3D viewer)
      2. 3D cell viewer (pre2018)
      3. Deep cell zoom (216,016 cells)
      Online modeling viewers
      1. Visual Guide to Human Cells
      2. Simularium (4D visual analysis)
      3. Integrated Mitotic Stem Cell
      4. └ Z-stack viewer
      5. └ 3D viewer
      6. Allen Integrated Cell viewer
      7. Label-free examples viewer
      8. 3D probabilistic model viewer
      Desktop tools
      1. Allen Cell & Structure Segmenter
      2. AGAVE 3D pathtrace image viewer
      Data & code
      1. Download cell data (images, genomics, features)
      2. Code repositories & software
  • Analysis & Modeling
      Allen Integrated Cell models
      1. Overview
      2. Integrated Mitotic Stem Cell
      3. └ Z-stack viewer
      4. └ 3D viewer
      5. Label-free Determination
      6. └ 3D viewer
      7. 3D Probabilistic Modeling
      8. └ 3D viewer
      9. Visual Guide to Human Cells
      4D biology models
      1. Simularium (online 4D viewer)
      Methodologies
      1. Drug perturbation pilot study
      2. hiPS cells during mitosis
      3. Differentiation into cardiomyocytes
  • Publications
      Articles
      1. All journal publications
      2. Preprints (biorxiv, arxiv)
      Posters
      1. Select posters
  • Education
      Education resources
      1. All Resources
      2. Teaching materials
      Online tools popular with teachers
      1. Visual Guide to Human Cells
      2. Integrated Mitotic Stem Cell
      3. Cell Feature Explorer (interactive plotting & 3D viewer)
      4. 3D cell viewer (pre2018 data)
      5. hiPS cell structure overview
  • Support
      Questions
      1. FAQs
      2. Forum
      Tutorials for digital tools
      1. Digital tool tutorials with videos
      2. Visual Guide tutorial
      3. AGAVE user guide
      Lab methods
      1. Instructional videos for success in the lab
      2. Standard operating procedures (written methods)
      3. Illustrated overviews
  • 🔍
      SEARCHBAR