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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.

Golgi visualized via sialyltransferase 1 (also known as ß-galactoside α-2,6-sialyltransferase1)

10/5/2017

 
A. Z-stack 1-copy edit
B. Z-stack 2-copy edit
C. Z-stack 2-copy edit
Figure 1. Z-stack movies of Golgi. A) Z-stack of live hiPS cells expressing one copy (monoallelic) of mEGFP-tagged sialyltransferase 1 (ST6GAL1) imaged on a spinning-disk confocal microscope. Movie starts at the bottom of the cells and ends at the top. B) Z-stack of live hiPS cells expressing two copies (biallelic) of mEGFP-tagged sialyltransferase 1 (ST6GAL1) imaged on a spinning-disk confocal microscope. Movie starts at the bottom of the cells and ends at the top. Contrast settings for these two movies are identical to highlight the increased signal in the biallelic edited line (right). C) Z-stack of live hiPS cells expressing two copies (biallelic) of mEGFP-tagged sialyltransferase 1 (ST6GAL1; white) stained for DNA (NucBlue Live; cyan) and plasma membrane (CellMask Deep Red; magenta). Movie starts at the bottom of the cells and ends at the top.
A. Timelapse 6 hours
B. Timelapse 100 minutes
Figure 2. Timelapse movies of Golgi. A) Timelapse movie of hiPS cells expressing two copies (biallelic) of mEGFP-tagged sialyltransferase 1 (ST6GAL1). Images were taken in 3D every 3 min for 6 hrs on a spinning-disk confocal microscope. Images are maximum intensity projections with the ‘despeckle’ ImageJ filter applied. Playback speed is 1800x real time. B) Timelapse movie of hiPS cells expressing two copies (biallelic) of mEGFP-tagged sialyltransferase 1 (ST6GAL1). Images were taken in 3D every 5 min for 100 min on a spinning-disk confocal microscope. Images are maximum intensity projections with the ‘despeckle’ ImageJ filter applied. Playback speed is 900x real time.
Observations
  • Sialyltransferase 1 is a transmembrane protein that resides within the Golgi apparatus. It catalyzes the transfer of sialic acid from an activated donor molecule to galactose.
  • In hiPS cells, the Golgi comprises a set of lamellar structures localized near the top of the cell, often partially covering the nucleus (see 3D reconstruction). The structure of the Golgi becomes more tubular as it descends downward into the cell.
  • During cell division, the Golgi fragments into numerous small foci that are dispersed throughout the cell, and excluded from the mitotic spindle. These Golgi fragments reassemble in daughter cells after division.
  • Golgi morphology is difficult to visualize in immunostained cells as its lamellar structure easily collapses into a tubular morphology in response to fixation.

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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
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      Tutorials for digital tools
      1. Digital tool tutorials with videos
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      3. AGAVE user guide
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