한빛사논문
- 조회346
Sheldon J. J. Kwok 1,*, Sarah Forward 1, Marissa D. Fahlberg 1, Emane Rose Assita 1, Sean Cosgriff 1, Seung Hyung Lee 1, Geoffrey R. Abbott 1, Han Zhu 1, Nicolas H. Minasian 1, A. Sean Vote 1, Nicola Martino 2 & Seok-Hyun Yun 2,*
1LASE Innovation Inc., Woburn, MA, USA.
2Harvard Medical School and Wellman Center for Photomedicine, Massachusetts General Hospital, Cambridge, MA, USA.
*Corresponding authors: correspondence to Sheldon J. J. Kwok or Seok-Hyun Yun
Abstract
Advances in immunology, immuno-oncology, drug discovery and vaccine development demand improvements in the capabilities of flow cytometry to allow it to measure more protein markers per cell at multiple timepoints. However, the size of panels of fluorophore markers is limited by overlaps in fluorescence-emission spectra, and flow cytometers typically perform cell measurements at one timepoint. Here we describe multi-pass high-dimensional flow cytometry, a method leveraging cellular barcoding via microparticles emitting near-infrared laser light to track and repeatedly measure each cell using more markers and fewer colours. By using live human peripheral blood mononuclear cells, we show that the method enables the time-resolved characterization of the same cells before and after stimulation, their analysis via a 10-marker panel with minimal compensation for spectral spillover and their deep immunophenotyping via a 32-marker panel, where the same cells are analysed in 3 back-to-back cycles with 10–13 markers per cycle, reducing overall spillover and simplifying marker-panel design. Cellular barcoding in flow cytometry extends the utility of the technique for high-dimensional multi-pass single-cell analyses.
논문정보
- Research article
- 2023년 11월 (BRIC 등록일 2024-01-16)
- 국외 연구진
댓글 작성 로그인
팝업 닫기관련 링크
