We report on the fabrication of multifunctional ratiometric probes for glucose and temperatures based on thermoresponsive poly(N-isopropylacrylamide) (PNIPAM) microgels covalently incorporated with glucose-recognizing moieties, N-acryloyl-3-aminophenylboronic acid (APBA), fluorescence resonance energy transfer (FRET) donor dyes, 4-(2-acryloyloxyethylamino)-7-nitro-2,1,3-benzoxadiazole (NBDAE), and rhodamine B-based FRET acceptors (RhBEA). P(NIPAM-APBA-NBDAE-RhBEA) microgels containing FRET pairs and APBA were synthesized via free radical emulsion copolymerization. The spatial proximity of FRET donors and acceptors within microgels can be tuned via thermo-induced microgel collapse or glucose-induced microgel swelling at appropriate pH and temperatures, leading to the facile modulation of FRET efficiencies. APBA moieties within P(NIPAM-APBA-NBDAE-RhBEA) microgels can bind with glucose at appropriate pH to form cyclic boronate moieties, which can decrease the pKa of APBA residues and increase the volume phas...