High-throughput spheroid screens using volume, resazurin reduction and acid phosphatase activity
Mainstream adoption of physiologically-relevant three-dimensional models has been slow in the last 50 years due to long, manual protocols with poor reproducibility, high price and closed commercial platforms. This chapter describes high-throughput, low-cost, open methods for spheroid viability asses...
| Main Authors: | , , |
|---|---|
| Other Authors: | |
| Format: | Book Section |
| Published: |
Humana Press (Springer)
2017
|
| Subjects: | |
| Online Access: | https://eprints.nottingham.ac.uk/43565/ |
| _version_ | 1848796715707006976 |
|---|---|
| author | Ivanov, Delyan Pavlov Grabowska, Anna M. Garnett, Martin C. |
| author2 | Gilbert, Daniel F. |
| author_facet | Gilbert, Daniel F. Ivanov, Delyan Pavlov Grabowska, Anna M. Garnett, Martin C. |
| author_sort | Ivanov, Delyan Pavlov |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | Mainstream adoption of physiologically-relevant three-dimensional models has been slow in the last 50 years due to long, manual protocols with poor reproducibility, high price and closed commercial platforms. This chapter describes high-throughput, low-cost, open methods for spheroid viability assessment which use readily-available reagents and open-source software to analyse spheroid volume, metabolism and enzymatic activity. We provide two ImageJ macros for automated spheroid size determination - for both single images and for images in stacks. We also share an Excel template spreadsheet allowing users to rapidly process spheroid size data, analyse plate uniformity (such as edge effects and systematic seeding errors), detect outliers and calculate dose-response. The methods would be useful to researchers in preclinical and translational research planning to move away from simplistic monolayer studies and explore 3D spheroid screens for drug safety and efficacy without substantial investment in money or time. |
| first_indexed | 2025-11-14T19:52:23Z |
| format | Book Section |
| id | nottingham-43565 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:52:23Z |
| publishDate | 2017 |
| publisher | Humana Press (Springer) |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-435652020-05-04T18:44:32Z https://eprints.nottingham.ac.uk/43565/ High-throughput spheroid screens using volume, resazurin reduction and acid phosphatase activity Ivanov, Delyan Pavlov Grabowska, Anna M. Garnett, Martin C. Mainstream adoption of physiologically-relevant three-dimensional models has been slow in the last 50 years due to long, manual protocols with poor reproducibility, high price and closed commercial platforms. This chapter describes high-throughput, low-cost, open methods for spheroid viability assessment which use readily-available reagents and open-source software to analyse spheroid volume, metabolism and enzymatic activity. We provide two ImageJ macros for automated spheroid size determination - for both single images and for images in stacks. We also share an Excel template spreadsheet allowing users to rapidly process spheroid size data, analyse plate uniformity (such as edge effects and systematic seeding errors), detect outliers and calculate dose-response. The methods would be useful to researchers in preclinical and translational research planning to move away from simplistic monolayer studies and explore 3D spheroid screens for drug safety and efficacy without substantial investment in money or time. Humana Press (Springer) Gilbert, Daniel F. Friedrich, Oliver 2017-05-04 Book Section PeerReviewed Ivanov, Delyan Pavlov, Grabowska, Anna M. and Garnett, Martin C. (2017) High-throughput spheroid screens using volume, resazurin reduction and acid phosphatase activity. In: Cell Viability Assays. Methods in Molecular Biology, 1601 . Humana Press (Springer), pp. 43-59. ISBN 978-1-4939-6960-9 alamar blue viability assays overlay culture hanging drop FiJi ImageJ image analysis three-dimensional cell culture in vitro model preclinical screening drug sensitivity https://link.springer.com/protocol/10.1007%2F978-1-4939-6960-9_4 doi:10.1007/978-1-4939-6960-9_4 doi:10.1007/978-1-4939-6960-9_4 |
| spellingShingle | alamar blue viability assays overlay culture hanging drop FiJi ImageJ image analysis three-dimensional cell culture in vitro model preclinical screening drug sensitivity Ivanov, Delyan Pavlov Grabowska, Anna M. Garnett, Martin C. High-throughput spheroid screens using volume, resazurin reduction and acid phosphatase activity |
| title | High-throughput spheroid screens using volume, resazurin reduction and acid phosphatase activity |
| title_full | High-throughput spheroid screens using volume, resazurin reduction and acid phosphatase activity |
| title_fullStr | High-throughput spheroid screens using volume, resazurin reduction and acid phosphatase activity |
| title_full_unstemmed | High-throughput spheroid screens using volume, resazurin reduction and acid phosphatase activity |
| title_short | High-throughput spheroid screens using volume, resazurin reduction and acid phosphatase activity |
| title_sort | high-throughput spheroid screens using volume, resazurin reduction and acid phosphatase activity |
| topic | alamar blue viability assays overlay culture hanging drop FiJi ImageJ image analysis three-dimensional cell culture in vitro model preclinical screening drug sensitivity |
| url | https://eprints.nottingham.ac.uk/43565/ https://eprints.nottingham.ac.uk/43565/ https://eprints.nottingham.ac.uk/43565/ |