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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- Text
- Journal contribution
- Biophysics
- Medicine
- Cell Biology
- Biotechnology
- Immunology
- Developmental Biology
- Infectious Diseases
- Virology
- Space Science
- Biological Sciences not elsewhere classified
- Chemical Sciences not elsewhere classified
- technique
- surface
- nanoparticle
- lanthanide bioprobes
- nontarget organisms
- method
- signal strength
- OSAM
- microscopy slides
- orthogonal scanning
- background noise