Automation of yeast spot assays using an affordable liquid handling robot
The spot assay of the budding yeast Saccharomyces cerevisiae is an experimental method that is used to evaluate the effect of genotypes, medium conditions, and environmental stresses on cell growth and survival. Automation of the spot assay experiments from preparing a dilution series to spotting to...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-04-01
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| Series: | SLAS Technology |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2472630322051871 |
| _version_ | 1797854480990994432 |
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| author | Shodai Taguchi Yasuyuki Suda Kenji Irie Haruka Ozaki |
| author_facet | Shodai Taguchi Yasuyuki Suda Kenji Irie Haruka Ozaki |
| author_sort | Shodai Taguchi |
| collection | DOAJ |
| description | The spot assay of the budding yeast Saccharomyces cerevisiae is an experimental method that is used to evaluate the effect of genotypes, medium conditions, and environmental stresses on cell growth and survival. Automation of the spot assay experiments from preparing a dilution series to spotting to observing spots continuously has been implemented based on large laboratory automation devices and robots, especially for high-throughput functional screening assays. However, there has yet to be an affordable solution for the automated spot assays suited to researchers in average laboratories and with high customizability for end-users. To make reproducible spot assay experiments widely available, we have automated the plate-based yeast spot assay of budding yeast using Opentrons OT-2 (OT-2), an affordable liquid-handling robot, and a flatbed scanner. We prepared a 3D-printed mount for the Petri dish to allow for precise placement of the Petri dish inside the OT-2. To account for the uneven height of the agar plates, which were made by human hands, we devised a method to adjust the z-position of the pipette tips based on the weight of each agar plate. During the incubation of the agar plates, a flatbed scanner was used to automatically take images of the agar plates over time, allowing researchers to quantify and compare the cell density within the spots at optimal time points a posteriori. Furthermore, the accuracy of the newly developed automated spot assay was verified by performing spot assays with human experimenters and the OT-2 and quantifying the yeast-grown area of the spots. This study will contribute to the introduction of automated spot assays and the automated acquisition of growth processes in conventional laboratories that are not adapted for high-throughput laboratory automation. |
| first_indexed | 2024-04-09T20:07:50Z |
| format | Article |
| id | doaj.art-a04b9af4eedd4916838b1b16b0fcc6c2 |
| institution | Directory Open Access Journal |
| issn | 2472-6303 |
| language | English |
| last_indexed | 2024-04-09T20:07:50Z |
| publishDate | 2023-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | SLAS Technology |
| spelling | doaj.art-a04b9af4eedd4916838b1b16b0fcc6c22023-04-01T08:51:01ZengElsevierSLAS Technology2472-63032023-04-012825562Automation of yeast spot assays using an affordable liquid handling robotShodai Taguchi0Yasuyuki Suda1Kenji Irie2Haruka Ozaki3Ph.D. Program in Humanics, School of Integrative and Global Majors, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8577, Japan; Department of Molecular Cell Biology, Faculty of Medicine, University of Tsukuba, Tsukuba, Japan, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8575, JapanDepartment of Molecular Cell Biology, Faculty of Medicine, University of Tsukuba, Tsukuba, Japan, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8575, Japan; Live Cell Super-Resolution Imaging Research Team, RIKEN Center for Advanced Photonics, Saitama 351-0198, JapanDepartment of Molecular Cell Biology, Faculty of Medicine, University of Tsukuba, Tsukuba, Japan, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8575, JapanBioinformatics Laboratory, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8575, Japan; Center for Artificial Intelligence Research, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8577, Japan; Corresponding author at: Bioinformatics Laboratory, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8575, Japan.The spot assay of the budding yeast Saccharomyces cerevisiae is an experimental method that is used to evaluate the effect of genotypes, medium conditions, and environmental stresses on cell growth and survival. Automation of the spot assay experiments from preparing a dilution series to spotting to observing spots continuously has been implemented based on large laboratory automation devices and robots, especially for high-throughput functional screening assays. However, there has yet to be an affordable solution for the automated spot assays suited to researchers in average laboratories and with high customizability for end-users. To make reproducible spot assay experiments widely available, we have automated the plate-based yeast spot assay of budding yeast using Opentrons OT-2 (OT-2), an affordable liquid-handling robot, and a flatbed scanner. We prepared a 3D-printed mount for the Petri dish to allow for precise placement of the Petri dish inside the OT-2. To account for the uneven height of the agar plates, which were made by human hands, we devised a method to adjust the z-position of the pipette tips based on the weight of each agar plate. During the incubation of the agar plates, a flatbed scanner was used to automatically take images of the agar plates over time, allowing researchers to quantify and compare the cell density within the spots at optimal time points a posteriori. Furthermore, the accuracy of the newly developed automated spot assay was verified by performing spot assays with human experimenters and the OT-2 and quantifying the yeast-grown area of the spots. This study will contribute to the introduction of automated spot assays and the automated acquisition of growth processes in conventional laboratories that are not adapted for high-throughput laboratory automation.http://www.sciencedirect.com/science/article/pii/S2472630322051871Automated pipettingYeast spot assayAffordable laboratory automationDrug resistanceAutomated image acquisition |
| spellingShingle | Shodai Taguchi Yasuyuki Suda Kenji Irie Haruka Ozaki Automation of yeast spot assays using an affordable liquid handling robot SLAS Technology Automated pipetting Yeast spot assay Affordable laboratory automation Drug resistance Automated image acquisition |
| title | Automation of yeast spot assays using an affordable liquid handling robot |
| title_full | Automation of yeast spot assays using an affordable liquid handling robot |
| title_fullStr | Automation of yeast spot assays using an affordable liquid handling robot |
| title_full_unstemmed | Automation of yeast spot assays using an affordable liquid handling robot |
| title_short | Automation of yeast spot assays using an affordable liquid handling robot |
| title_sort | automation of yeast spot assays using an affordable liquid handling robot |
| topic | Automated pipetting Yeast spot assay Affordable laboratory automation Drug resistance Automated image acquisition |
| url | http://www.sciencedirect.com/science/article/pii/S2472630322051871 |
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