Polymerization-induced self-assembly (PISA) is used for the highly convenient and efficient preparation of ampholytic diblock copolymer nanoparticles directly in acidic aqueous solution. Cationic nanoparticles comprising a protonated polyamine stabilizer block and a hydrophobic polyacid core-forming block are formed at pH 2. Micelle inversion occurs at pH 10 to produce anionic nanoparticles with an ionized polyacid stabilizer block and a hydrophobic polyamine core-forming block. Macroscopic precipitation occurs at around pH 6-7, which lies close to the isoelectric point of this ampholytic diblock copolymer. Incorporation of fluorescein and rhodamine dye labels into the acid and amine blocks, respectively, leads to dual-color bifluorescent self-reporting pH-responsive nanoparticles.