Comparison of Cell Arrays and Multi-Well Plates in Microscopy-Based Screening
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture and Materials
2.2. Preparation of on Cell Arrays for Small Interfering RNA Transfection
2.3. Coating of Multi-Well Plates with Small Interfering RNAs
2.4. Cell Cycle Progression Assay
2.5. Epidermal Growth Factor Internalization Assay
2.6. Microscopy
2.7. Image Analysis
2.8. Statistical Data Analysis
3. Results
3.1. Set-Up of the Assay to Quantify the Arrest of Cell Cycle Progression
3.2. Evaluation of the Assay for Cell Cycle Progression
3.3. Evaluation of the Epidermal Growth Factor Endocytosis Assay
4. Discussion
Supplementary Materials
Author Contributions
Acknowledgments
Conflicts of Interest
References
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Feature | Cell Arrays | Multi-Well Plates |
---|---|---|
High-throughput | yes | yes/no |
Genetic screens | yes | yes |
Chemical screens | yes/no | yes |
Reverse transfection | yes | yes |
Direct transfection | yes/no | yes |
Assay performance needs robotics | no | yes |
Edge effects | no | yes |
Complex assays can be performed | yes | yes/no |
Assay can be performed in cells which are available in small numbers | yes | no |
Small amounts of the reagents are required for screening | yes | no |
Cell migration can be analysed | no | yes |
Cells with the extended shape (e.g., neurons) can be analysed | no | yes |
Many cells are available for statistics from one replicate | no | yes |
Possibility to cross-contaminate the samples | yes/no | no |
Comparison | Well-Plates | Cell Arrays | ||
---|---|---|---|---|
Mean Phenotype | p-Value | Mean Phenotype | p-Value | |
Spindle phenotype | ||||
NC | 8.6 ± 1.2% | 9.7 ± 1.9% | ||
KIF11 | 54.8 ± 5.3% | 6.9 × 10−15 | 28.3 ± 7.4% | 2.6 × 10−9 |
PLK1 | 66.4 ± 7.6% | 1.6 × 10−15 | 29.2 ± 7.9% | 5.1 × 10−7 |
INCENP | 18.9 ± 3.3% | 1.5 × 10−9 | 13.4 ± 6.6% | 3.9 × 10−2 |
Cytokinesis phenotype | ||||
NC | 5.9 ± 1.2% | 8 ± 3.9% | ||
KIF11 | 7.8 ± 2% | 1 | 11.5 ± 5.6% | 3 × 10−2 |
PLK1 | 10.5 ± 1.5% | 2 × 10−1 | 6.6 ± 5.4% | 9.7 × 10−1 |
INCENP | 35.5 ± 10.1% | 2.6 × 10−6 | 32.7 ± 13.1% | 1.8 × 10−6 |
Comparison | Well-Plates | Cell Arrays | ||
---|---|---|---|---|
Mean Cell Numbers | p-Value | Mean Cell Numbers | p-Value | |
NC | 100% | 100% | ||
PLK1 | 49.6% | 9.9 × 10−9 | 68.2% | 1.2 × 10−2 |
KIF11 | 58.9% | 8.2 × 10−7 | 73.7% | 2.2 × 10−2 |
INCENP | 59.2% | 4.6 × 10−4 | 93.1% | 9.1 × 10−1 |
Comparison | Well-Plates | Cell Arrays | ||
---|---|---|---|---|
Mean Intracellular EGF Signal | p-Value | Mean Intracellular EGF Signal | p-Value | |
NC | 89.8 ± 5% | 91.8 ± 3.3% | ||
EGFR_1 | 44.4 ± 12.8% | 8.8 × 10−14 | 38.7 ± 6.6% | 4.1 × 10−14 |
EGFR_2 | 33.5 ± 10.8% | 1.5 × 10−14 | 45.9 ± 8.4% | 8.8 × 10−14 |
EGFR_3 | 28 ± 10.8% | 4.4 × 10−10 | 33 ± 5.5% | 4.7 × 10−14 |
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Becker, A.-K.; Erfle, H.; Gunkel, M.; Beil, N.; Kaderali, L.; Starkuviene, V. Comparison of Cell Arrays and Multi-Well Plates in Microscopy-Based Screening. High-Throughput 2018, 7, 13. https://doi.org/10.3390/ht7020013
Becker A-K, Erfle H, Gunkel M, Beil N, Kaderali L, Starkuviene V. Comparison of Cell Arrays and Multi-Well Plates in Microscopy-Based Screening. High-Throughput. 2018; 7(2):13. https://doi.org/10.3390/ht7020013
Chicago/Turabian StyleBecker, Ann-Kristin, Holger Erfle, Manuel Gunkel, Nina Beil, Lars Kaderali, and Vytaute Starkuviene. 2018. "Comparison of Cell Arrays and Multi-Well Plates in Microscopy-Based Screening" High-Throughput 7, no. 2: 13. https://doi.org/10.3390/ht7020013