Journal Club: Expansion Microscopy 06/19/15
Chen, Tillberg and Boyden, Expansion Microscopy
- presented by Yari
- label sample
- gelation in polyectrolye network
- proteolysis tissue / polymer sample
- expand in water
- 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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