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Ultrafast immunostaining of organ-scale tissues for scalable proteomic phenotyping

Dae Hee Yun, Young-Gyun Park, Jae Hun Cho, Lee Kamentsky, Nicholas B. Evans, Alex Albanese, Katherine Xie, Justin Swaney, Chang Ho Sohn, Yuxuan Tian, Qiangge Zhang, Gabi Drummond, Webster Guan, Nicholas DiNapoli, Heejin Choi, Hae-Yoon Jung, Luzdary Ruelas, Guoping Feng, View ORCID ProfileKwanghun Chung
doi: https://doi.org/10.1101/660373
Dae Hee Yun
1Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
2Picower Institute for Learning and Memory, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
3Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
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Young-Gyun Park
1Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
2Picower Institute for Learning and Memory, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
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Jae Hun Cho
4Department of Chemical Engineering, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
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Lee Kamentsky
1Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
2Picower Institute for Learning and Memory, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
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Nicholas B. Evans
1Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
2Picower Institute for Learning and Memory, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
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Alex Albanese
1Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
2Picower Institute for Learning and Memory, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
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Katherine Xie
1Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
2Picower Institute for Learning and Memory, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
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Justin Swaney
4Department of Chemical Engineering, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
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Chang Ho Sohn
1Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
2Picower Institute for Learning and Memory, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
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Yuxuan Tian
4Department of Chemical Engineering, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
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Qiangge Zhang
2Picower Institute for Learning and Memory, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
5McGovern Institute for Brain Research, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
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Gabi Drummond
3Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
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Webster Guan
4Department of Chemical Engineering, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
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Nicholas DiNapoli
1Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
2Picower Institute for Learning and Memory, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
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Heejin Choi
1Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
2Picower Institute for Learning and Memory, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
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Hae-Yoon Jung
1Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
2Picower Institute for Learning and Memory, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
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Luzdary Ruelas
4Department of Chemical Engineering, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
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Guoping Feng
2Picower Institute for Learning and Memory, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
5McGovern Institute for Brain Research, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
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Kwanghun Chung
1Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
2Picower Institute for Learning and Memory, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
3Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
4Department of Chemical Engineering, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA
6Broad Institute of Harvard University and MIT, Cambridge, MA, USA
7Yonsei-IBS Institute, Yonsei University, Seoul 03722, Republic of Korea
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  • ORCID record for Kwanghun Chung
  • For correspondence: khchung@mit.edu
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ABSTRACT

Studying the function and dysfunction of complex biological systems necessitates comprehensive understanding of individual cells. Advancements in three-dimensional (3D) tissue processing and imaging modalities have enabled rapid visualization and phenotyping of cells in their spatial context. However, system-wide interrogation of individual cells within large intact tissue remains challenging, low throughput, and error-prone owing to the lack of robust labeling technologies. Here we introduce a rapid, versatile, and scalable method, eFLASH, that enables complete and uniform labeling of organ-scale tissue within one day. eFLASH dynamically modulates chemical transport and reaction kinetics to establish system-wide uniform labeling conditions throughout the day-long labeling period. This unique approach enables the same protocol to be compatible with a wide range of tissue types and probes, enabling combinatorial molecular phenotyping across different organs and species. We applied eFLASH to generate quantitative maps of various cell types in mouse brains. We also demonstrated multidimensional cell profiling in a marmoset brain block. We envision that eFLASH will spur holistic phenotyping of emerging animal models and disease models to help assess their functions and dysfunctions.

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Posted June 05, 2019.
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Ultrafast immunostaining of organ-scale tissues for scalable proteomic phenotyping
Dae Hee Yun, Young-Gyun Park, Jae Hun Cho, Lee Kamentsky, Nicholas B. Evans, Alex Albanese, Katherine Xie, Justin Swaney, Chang Ho Sohn, Yuxuan Tian, Qiangge Zhang, Gabi Drummond, Webster Guan, Nicholas DiNapoli, Heejin Choi, Hae-Yoon Jung, Luzdary Ruelas, Guoping Feng, Kwanghun Chung
bioRxiv 660373; doi: https://doi.org/10.1101/660373
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Ultrafast immunostaining of organ-scale tissues for scalable proteomic phenotyping
Dae Hee Yun, Young-Gyun Park, Jae Hun Cho, Lee Kamentsky, Nicholas B. Evans, Alex Albanese, Katherine Xie, Justin Swaney, Chang Ho Sohn, Yuxuan Tian, Qiangge Zhang, Gabi Drummond, Webster Guan, Nicholas DiNapoli, Heejin Choi, Hae-Yoon Jung, Luzdary Ruelas, Guoping Feng, Kwanghun Chung
bioRxiv 660373; doi: https://doi.org/10.1101/660373

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