ThelocalCa^2+releasefromtheheterogeneouslydistributedendoplasmicreticulum(ER)calciumstorehasacriticalroleincalciumhomeostasisandcellularfunction.However;singlefluorescentproteinbasedERcalciumprobesexperiencechallengesinquantifyingtheERcalciumstoreindifferinglivecells,andintensity-basedmeasurementsmakeitdifficulttodetectlocalcalciummicrodomainsintheER.Here,wedevelopedageneticallyencodedratiometricERcalciumindicator[GCEPIA1-SNAPer]thatcandetectthereal-timeERcalciumstoreandlocalcalciummicrodomainsinlivecells.GCEPIA1-SNAPerwaslocatedinthelumenoftheERandshowedalinear;reversibleandrapidresponsetochangesintheERcalciumstore.TheGCEPIA1-SNAPerprobeeffectivelymonitoredthedepletionoftheERcalciumstorebyTGorstarvationtreatment,andthrough让suseweidentifiedheterogeneouslydistributedcalciummicrodomainsintheERwhichwerecorrelatedw让hthedistributionofSTIM1clustersuponERcalciumstoredepletion.Lastly,GCEPIA1-SNAPercanbeusedtodetecttheERcalciumstorebyhigh-throughputflowcytometryandconferstheabilitytostudythefunctionofcalciummicrodomainsoftheER.
