Critical Evaluation of Techniques to Detect and Measure Cell Death – Study in a Model of UV Radiation of the Leukaemic Cell Line HL60
The reliability of eight distinct methods (Giemsa staining, trypan blue exclusion, acridine orange/ethidium bromide (AO/EB) double staining for fluorescence microscopy and flow cytometry, propidium iodide (PI) staining, annexin V assay, TUNEL assay and DNA ladder) for detection and quantification of...
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| Format: | Article |
| Language: | English |
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Wiley
1999-01-01
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| Series: | Analytical Cellular Pathology |
| Online Access: | http://dx.doi.org/10.1155/1999/176515 |
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| _version_ | 1850167592801533952 |
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| author | Marina Leite Margarida Quinta‐Costa Pedro Simas Leite José Eduardo Guimarães |
| author_facet | Marina Leite Margarida Quinta‐Costa Pedro Simas Leite José Eduardo Guimarães |
| author_sort | Marina Leite |
| collection | DOAJ |
| description | The reliability of eight distinct methods (Giemsa staining, trypan blue exclusion, acridine orange/ethidium bromide (AO/EB) double staining for fluorescence microscopy and flow cytometry, propidium iodide (PI) staining, annexin V assay, TUNEL assay and DNA ladder) for detection and quantification of cell death (apoptosis and necrosis) was evaluated and compared. Each of these methods detects different morphological or biochemical features of these two processes. The comparative analysis of the 8 techniques revealed that AO/EB (read in fluorescence microscopy) provides a reliable method to measure cells in different compartments (or pathways) of cell death though it is very time consuming. PI staining and TUNEL assay were also sensitive in detecting very early signs of apoptosis, but do not allow precise quantification of apoptotic cells. These three methods were concordant in relation to induction of apoptosis and necrosis in HL60 cells with the various UV irradiation time periods tested. Both AO/EB (read by flow cytometry) and annexin V‐FITC/PI failed to detect the same number of early apoptotic cells as the other three methods. Trypan blue is valueless for this purpose. Giemsa and DNA ladder might be useful as confirmatory tests in some situations. |
| format | Article |
| id | doaj-art-e7a144da326e4eb682ac42bb8f33e643 |
| institution | OA Journals |
| issn | 0921-8912 1878-3651 |
| language | English |
| publishDate | 1999-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Analytical Cellular Pathology |
| spelling | doaj-art-e7a144da326e4eb682ac42bb8f33e6432025-08-20T02:21:10ZengWileyAnalytical Cellular Pathology0921-89121878-36511999-01-01193-413915110.1155/1999/176515Critical Evaluation of Techniques to Detect and Measure Cell Death – Study in a Model of UV Radiation of the Leukaemic Cell Line HL60Marina Leite0Margarida Quinta‐Costa1Pedro Simas Leite2José Eduardo Guimarães3IPATIMUP, Instituto de Patologia e Imunologia Molecular da Universidade do Porto, PortugalIPATIMUP, Instituto de Patologia e Imunologia Molecular da Universidade do Porto, PortugalIPATIMUP, Instituto de Patologia e Imunologia Molecular da Universidade do Porto, PortugalIPATIMUP, Instituto de Patologia e Imunologia Molecular da Universidade do Porto, PortugalThe reliability of eight distinct methods (Giemsa staining, trypan blue exclusion, acridine orange/ethidium bromide (AO/EB) double staining for fluorescence microscopy and flow cytometry, propidium iodide (PI) staining, annexin V assay, TUNEL assay and DNA ladder) for detection and quantification of cell death (apoptosis and necrosis) was evaluated and compared. Each of these methods detects different morphological or biochemical features of these two processes. The comparative analysis of the 8 techniques revealed that AO/EB (read in fluorescence microscopy) provides a reliable method to measure cells in different compartments (or pathways) of cell death though it is very time consuming. PI staining and TUNEL assay were also sensitive in detecting very early signs of apoptosis, but do not allow precise quantification of apoptotic cells. These three methods were concordant in relation to induction of apoptosis and necrosis in HL60 cells with the various UV irradiation time periods tested. Both AO/EB (read by flow cytometry) and annexin V‐FITC/PI failed to detect the same number of early apoptotic cells as the other three methods. Trypan blue is valueless for this purpose. Giemsa and DNA ladder might be useful as confirmatory tests in some situations.http://dx.doi.org/10.1155/1999/176515 |
| spellingShingle | Marina Leite Margarida Quinta‐Costa Pedro Simas Leite José Eduardo Guimarães Critical Evaluation of Techniques to Detect and Measure Cell Death – Study in a Model of UV Radiation of the Leukaemic Cell Line HL60 Analytical Cellular Pathology |
| title | Critical Evaluation of Techniques to Detect and Measure Cell Death – Study in a Model of UV Radiation of the Leukaemic Cell Line HL60 |
| title_full | Critical Evaluation of Techniques to Detect and Measure Cell Death – Study in a Model of UV Radiation of the Leukaemic Cell Line HL60 |
| title_fullStr | Critical Evaluation of Techniques to Detect and Measure Cell Death – Study in a Model of UV Radiation of the Leukaemic Cell Line HL60 |
| title_full_unstemmed | Critical Evaluation of Techniques to Detect and Measure Cell Death – Study in a Model of UV Radiation of the Leukaemic Cell Line HL60 |
| title_short | Critical Evaluation of Techniques to Detect and Measure Cell Death – Study in a Model of UV Radiation of the Leukaemic Cell Line HL60 |
| title_sort | critical evaluation of techniques to detect and measure cell death study in a model of uv radiation of the leukaemic cell line hl60 |
| url | http://dx.doi.org/10.1155/1999/176515 |
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