As　the　reliable　biomarkers　to　evaluate　the　diabetes　and　neurological　disease,　sensitive　and　accurate　detection　of　glucose　and　glutathione　(GSH)　in　biological　samples　is　necessary　for　early　precaution　and　diagnosis　of　related-diseases.　The　single　red　upconversion　nanoparticles　(UCNPs)　especially　with　core-shell　structure　can　penetrate　deeper　biological　tissues　and　cause　less　energy　loss　and　thus　have　higher　sensitivity　and　accuracy.　Additionally,　an　enzyme-controlled　cascade　signal　amplification　(ECSAm)　strategy　will　further　enhance　sensitivity.　Herein,　using　single　red　UCNPs　with　core-shell　structure　as　the　luminescent　material,　a　fluorescent　sensor　based　on　ECSAm　was　developed　for　the　highly　sensitive　and　accurate　detection　of　glucose　and　GSH.　Under　the　optimal　conditions,　the　limits　of　detection　for　glucose　and　GSH　by　fluorescent　method　were　0.03　μM　and　0.075　μM,　separately.　This　assay　was　used　to　analyze　the　content　of　glucose　and　GSH　in　serum　samples,　and　the　obtained　data　was　close　to　that　of　commercial　blood　glucose　and　GSH　detection　kit.　The　developed　sensor　platform　based　on　single　red　UCNPs　with　core-shell　structure　and　ECSAm　can　be　a　promising　method　for　the　accurate　and　sensitive　detection　of　glucose　and　GSH　in　biological　samples.　©　2024　Elsevier　B.V.