Journal Club: Expansion Microscopy 06/19/15

Chen, Tillberg and Boyden, Expansion Microscopy

  • presented by Yari
  • approach
    • label sample
    • gelation in polyectrolye network
    • proteolysis tissue / polymer sample
    • expand in water
    • image
  • super absorbant polymers (used in diapers)
    • sodium acrolyate + acrylomide
  • special probes required. Can’t just use labeled antibodies (they’ll get eaten)
    • add a single-stranded oligo-nucleotide to antibody. Corresponding oligo with dye and polymer attaching group (methacryloyl) hybridized to this. Thus it sticks to the gel.
  • primarily getting linear expansion factor of 4. Test up to factor of 9 (possibly at cost of stability).
  • initial polymer is highly crumpled because its in 2M salt. Expansion is pure water.
  • rescaled expanded sample, looks same before and after expansion. Nice.
  • compare SIM images before and after. claim ~60 nm resolution
  • see some non-uniform distoration. On order of distoration of PSF of confocal. Mostly a problem only at very large distances. (could correct for this with a few embedded fiducials)
  • clathrin pits look good too.
  • tissue is extremely clear. (detergent)
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