Fig. 1: Development of BOOST workflow.
From: BOOST: a robust ten-fold expansion method on hour-scale
A Optimized BOOST microwave-assisted protocol for three sample types. Reagents and processing times are listed for each step. B Comparison of processing times for selected ExM techniques for three types of samples. C Microwave-assisted BOOST hydrogel chemistry uses free-radical initiated polymerization of acrylamide (AA) and N,N’-Methylenebis(acrylamide) (bis), followed by alkaline hydrolysis to convert amide groups into charged carboxyl groups, transforming the gel into a swellable polyelectrolyte hydrogel. D Microwave-assisted anchoring with the reactions between the amine groups of the biomolecules and aldehyde of PFA to produce methylol groups, followed by reactions with AA to be anchored to the hydrogel. E A schematic diagram of biomolecules undergoing microwave-assisted gelation, hydrolysis, denaturation, and expansion; green dots and purple sticks depict the anchorable gel-reactive groups on biomolecules. F Optimization of concentrations of AA and bis for maximizing the expansion factor (EF) under 20-minute microwave-assisted hydrolysis (mean ± SD, n = 3 from three independent experiments). G Gel mechanical performance measurement showing the deformation indexes of BOOST gel formulations and the TREx gel. Squares in the background indicate 5 mm × 5 mm areas. Additional note: graphical icons in (1A, 1C–E) are created using Biorender; source data are provided as a Source Data file.
