TY - GEN
T1 - Quality control of highly multiplexed proteomic immunostaining with quantum dots
T2 - 31st Annual International Conference of the IEEE Engineering in Medicine and Biology Society: Engineering the Future of Biomedicine, EMBC 2009
AU - Moffitt, Richard A.
AU - Caldwell, Matthew L.
AU - Liu, Tao
AU - Liu, Jian
AU - Nie, Shuming
AU - Wang, May D.
PY - 2009
Y1 - 2009
N2 - The process of developing molecular assays for disease diagnosis and prognosis requires cross-disciplinary research which monitors quality and reproducibility at all levels. This paper discusses challenges in the quality control of highly multiplexed Quantum Dot (QD) staining and provides a method for improving accuracy of QD quantification in two phases. Phase one is the estimation of unintended crosstalk between multiplexed QD-antibody reporters, and phase two is digital correction of this crosstalk. Results show that crosstalk varies among tissues and reagents, and in some cases it can be on the same order of magnitude as the original intended signal. In cases where target protein expression is assumed to be independent, crosstalk can be empirically estimated from imaging data and corrected for. This work is expected to improve the overall reproducibility and quantification of multiplexed QD staining.
AB - The process of developing molecular assays for disease diagnosis and prognosis requires cross-disciplinary research which monitors quality and reproducibility at all levels. This paper discusses challenges in the quality control of highly multiplexed Quantum Dot (QD) staining and provides a method for improving accuracy of QD quantification in two phases. Phase one is the estimation of unintended crosstalk between multiplexed QD-antibody reporters, and phase two is digital correction of this crosstalk. Results show that crosstalk varies among tissues and reagents, and in some cases it can be on the same order of magnitude as the original intended signal. In cases where target protein expression is assumed to be independent, crosstalk can be empirically estimated from imaging data and corrected for. This work is expected to improve the overall reproducibility and quantification of multiplexed QD staining.
UR - http://www.scopus.com/inward/record.url?scp=77951014588&partnerID=8YFLogxK
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U2 - 10.1109/IEMBS.2009.5332857
DO - 10.1109/IEMBS.2009.5332857
M3 - Conference contribution
C2 - 19963937
AN - SCOPUS:77951014588
SN - 9781424432967
T3 - Proceedings of the 31st Annual International Conference of the IEEE Engineering in Medicine and Biology Society: Engineering the Future of Biomedicine, EMBC 2009
SP - 6739
EP - 6742
BT - Proceedings of the 31st Annual International Conference of the IEEE Engineering in Medicine and Biology Society
PB - IEEE Computer Society
Y2 - 2 September 2009 through 6 September 2009
ER -