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Re-embedding in neutral hydrogel

OM Ons M’Saad
JB Joerg Bewersdorf
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Expanded hydrogels were incubated in a fresh re-embedding neutral gel solution (10% (w/v) AAm + 0.05% (w/v) DHEBA + 0.05% (v/v) TEMED + 0.05% (w/v) APS in 1× PBS) three times for 20 min each on a rocking platform at RT. Immediately after, residual gel solution was removed by extensive but gentle pressing with Kimwipes. The gels were then sandwiched between two pieces of no. 1.5 coverslips and incubated at 37 °C for 1.5 h in a nitrogen-filled humidified chamber. Next, the gels were detached from the coverslip and washed three times with 1× PBS for 30 min each on a rocking platform at RT. Gels were incubated in post-fix solution (0.7% FA + 1% (w/v) AAm in 1× PBS) for 15 min at RT and then for 6–9 h at 37 °C. The gels were subsequently washed three times with 1× PBS for 30 min each on a rocking platform at RT.

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