Label-free redox imaging of patient-derived organoids using selective plane illumination microscopy

Peter F Favreau; Jiaye He; Daniel A Gil; Dustin A Deming; Jan Huisken; Melissa C Skala

doi:10.1364/BOE.389164

Label-free redox imaging of patient-derived organoids using selective plane illumination microscopy

Biomed Opt Express. 2020 Apr 16;11(5):2591-2606. doi: 10.1364/BOE.389164. eCollection 2020 May 1.

Authors

Peter F Favreau¹, Jiaye He^{1

2

3}, Daniel A Gil^{1

4}, Dustin A Deming^{5

6

7

8}, Jan Huisken^{1

3}, Melissa C Skala^{1

4}

Affiliations

¹ Morgridge Institute for Research, Madison, WI 53715, USA.
² Max Planck Institute for Molecular Cell Biology and Genetics, 01307 Dresden, Saxony, Germany.
³ Department of Integrative Biology, University of Wisconsin-Madison, Madison, WI 53715, USA.
⁴ Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53715, USA.
⁵ University of Wisconsin Carbone Cancer Center, Madison, WI 53715, USA.
⁶ Division of Hematology and Oncology, Department of Medicine, University of Wisconsin-Madison, Madison, WI 53715, USA.
⁷ McArdle Laboratory for Cancer Research, Department of Oncology, University of Wisconsin-Madison, Madison, WI 53715, USA.
⁸ William S Middleton Memorial Veterans Hospital, Madison, WI 53715, USA.

Abstract

High-throughput drug screening of patient-derived organoids offers an attractive platform to determine cancer treatment efficacy. Here, selective plane illumination microscopy (SPIM) was used to determine treatment response in organoids with endogenous fluorescence from the metabolic coenzymes NAD(P)H and FAD. Rapid 3-D autofluorescence imaging of colorectal cancer organoids was achieved. A quantitative image analysis approach was developed to segment each organoid and quantify changes in endogenous fluorescence caused by treatment. Quantitative analysis of SPIM volumes confirmed the sensitivity of patient-derived organoids to standard therapies. This proof-of-principle study demonstrates that SPIM is a powerful tool for high-throughput screening of organoid treatment response.

Abstract

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