title: Resolving low-expression cell surface antigens by time-gated orthogonal scanning automated microscopy.
abstract: We report a highly sensitive method for rapid identification and quantification of rare-event cells carrying low-abundance surface biomarkers. The method applies lanthanide bioprobes and time-gated detection to effectively eliminate both nontarget organisms and background noise and utilizes the europium containing nanoparticles to further amplify the signal strength by a factor of ∼20. Of interest is that these nanoparticles did not correspondingly enhance the intensity of nonspecific binding. Thus, the dramatically improved signal-to-background ratio enables the low-expression surface antigens on single cells to be quantified. Furthermore, we applied an orthogonal scanning automated microscopy (OSAM) technique to rapidly process a large population of target-only cells on microscopy slides, leading to quantitative statistical data with high certainty. Thus, the techniques together resolved nearly all false-negative events from the interfering crowd including many false-positive events.
authors.forename: Jie
authors.initials: J
authors.lastname: Lu
authors.forename: Jody
authors.initials: J
authors.lastname: Martin
authors.forename: Yiqing
authors.initials: Y
authors.lastname: Lu
authors.forename: Jiangbo
authors.initials: J
authors.lastname: Zhao
authors.forename: Jingli
authors.initials: J
authors.lastname: Yuan
authors.forename: Martin
authors.initials: M
authors.lastname: Ostrowski
authors.forename: Ian
authors.initials: I
authors.lastname: Paulsen
authors.forename: James A
authors.initials: JA
authors.lastname: Piper
authors.forename: Dayong
authors.initials: D
authors.lastname: Jin
mesh_terms: D018952:Antigens, CD34 / Q000378:metabolism*; D001331:Automation; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006801:Humans; D008853:Microscopy / Q000379:methods*; D013997:Time Factors
chemical_list: D018952:Antigens, CD34