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Cell structure observations

Observations about microscopy videos for 16 of the 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. Our science: CellScapes
      2. Past foundational projects
      3. News feed
      4. About us
      5. Careers
  • Allen Cell Collection
      Order cells & plasmids
      1. Cell Catalog
      2. Disease Collection Cell Catalog
      3. Cell Catalog quickview
      4. Cell video shorts
      Lab methods
      1. Video protocols
      2. Written protocols
      3. Our methodology
      4. Support forum
      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
      General
      1. Tools & resources overview
      2. Download cell data (images, genomics, features)
      3. Code repositories & software
      Desktop tools
      1. Allen Cell & Structure Segmenter
      2. AGAVE 3D pathtrace image viewer
      Web tools
      1. BioFile Finder
      2. Cell Feature Explorer
      3. Integrated Mitotic Stem Cell
      4. └ Z-stack viewer
      5. └ 3D viewer
      Web tools (con't)
      1. Simularium viewer
      2. Timelapse Feature Explorer
      3. Visual Guide to Human Cells
      4. Vol-E (Web Volume Viewer)
      5. 3D Cell Viewer
  • Analysis & Modeling
      Allen Integrated Cell models
      1. Visual Guide to Human Cells
      2. Integrated Mitotic Stem Cell
      3. └ Z-stack viewer
      4. └ 3D viewer
      5. Allen Integrated Cell
      6. └ 3D Probabilistic Modeling
      7. └ Label-free Determination
      4D biology models
      1. Simularium viewer
      Methodologies
      1. Drug perturbation pilot study
      2. hiPS cells during mitosis
      3. Differentiation into cardiomyocytes
  • Publications
      Articles
      1. Publications
      2. Preprints
      Presentations
      1. Talks & posters
  • Education
      Educational resources
      1. All resources
      2. Teaching materials
      Online tools popular with teachers
      1. Visual Guide to Human Cells
      2. Integrated Mitotic Stem Cell
      3. 3D Cell Feature Explorer
      4. 3D Cell Viewer
      5. hiPS cell structure overview
  • Support
      Questions
      1. FAQs
      2. Forum
      Tutorials for digital tools
      1. Video tutorials
      2. Visual Guide tutorial
      3. AGAVE documentation
      Lab methods
      1. Video protocols
      2. Written protocols
      3. Our methodology
  • 🔍
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