Whole-Cell Multiparameter Assay for Ricin and Abrin Activity-Based Digital Holographic Microscopy
Ricin and abrin are ribosome-inactivating proteins leading to inhibition of protein synthesis and cell death. These toxins are considered some of the most potent and lethal toxins against which there is no available antidote. Digital holographic microscopy (DHM) is a time-lapse, label-free, and noni...
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| Format: | Article |
| Language: | English |
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MDPI AG
2019-03-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/11/3/174 |
| _version_ | 1798005047044341760 |
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| author | Efi Makdasi Orly Laskar Elad Milrot Ofir Schuster Shlomo Shmaya Shmuel Yitzhaki |
| author_facet | Efi Makdasi Orly Laskar Elad Milrot Ofir Schuster Shlomo Shmaya Shmuel Yitzhaki |
| author_sort | Efi Makdasi |
| collection | DOAJ |
| description | Ricin and abrin are ribosome-inactivating proteins leading to inhibition of protein synthesis and cell death. These toxins are considered some of the most potent and lethal toxins against which there is no available antidote. Digital holographic microscopy (DHM) is a time-lapse, label-free, and noninvasive imaging technique that can provide phase information on morphological features of cells. In this study, we employed DHM to evaluate the morphological changes of cell lines during ricin and abrin intoxication. We showed that the effect of these toxins is characterized by a decrease in cell confluence and changes in morphological parameters such as cell area, perimeter, irregularity, and roughness. In addition, changes in optical parameters such as phase-shift, optical thickness, and effective-calculated volume were observed. These effects were completely inhibited by specific neutralizing antibodies. An enhanced intoxication effect was observed for preadherent compared to adherent cells, as was detected in early morphology changes and confirmed by annexin V/propidium iodide (PI) apoptosis assay. Detection of the dynamic changes in cell morphology at initial stages of cell intoxication by DHM emphasizes the highly sensitive and rapid nature of this method, allowing the early detection of active toxins. |
| first_indexed | 2024-04-11T12:34:11Z |
| format | Article |
| id | doaj.art-1b4182ec92fb49d5a020704351d8839f |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-04-11T12:34:11Z |
| publishDate | 2019-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-1b4182ec92fb49d5a020704351d8839f2022-12-22T04:23:41ZengMDPI AGToxins2072-66512019-03-0111317410.3390/toxins11030174toxins11030174Whole-Cell Multiparameter Assay for Ricin and Abrin Activity-Based Digital Holographic MicroscopyEfi Makdasi0Orly Laskar1Elad Milrot2Ofir Schuster3Shlomo Shmaya4Shmuel Yitzhaki5The Department of Infectious Diseases, Israel Institute for Biological Research, Ness-Ziona 74100, IsraelThe Department of Infectious Diseases, Israel Institute for Biological Research, Ness-Ziona 74100, IsraelThe Department of Infectious Diseases, Israel Institute for Biological Research, Ness-Ziona 74100, IsraelThe Department of Infectious Diseases, Israel Institute for Biological Research, Ness-Ziona 74100, IsraelThe Department of Infectious Diseases, Israel Institute for Biological Research, Ness-Ziona 74100, IsraelThe Department of Infectious Diseases, Israel Institute for Biological Research, Ness-Ziona 74100, IsraelRicin and abrin are ribosome-inactivating proteins leading to inhibition of protein synthesis and cell death. These toxins are considered some of the most potent and lethal toxins against which there is no available antidote. Digital holographic microscopy (DHM) is a time-lapse, label-free, and noninvasive imaging technique that can provide phase information on morphological features of cells. In this study, we employed DHM to evaluate the morphological changes of cell lines during ricin and abrin intoxication. We showed that the effect of these toxins is characterized by a decrease in cell confluence and changes in morphological parameters such as cell area, perimeter, irregularity, and roughness. In addition, changes in optical parameters such as phase-shift, optical thickness, and effective-calculated volume were observed. These effects were completely inhibited by specific neutralizing antibodies. An enhanced intoxication effect was observed for preadherent compared to adherent cells, as was detected in early morphology changes and confirmed by annexin V/propidium iodide (PI) apoptosis assay. Detection of the dynamic changes in cell morphology at initial stages of cell intoxication by DHM emphasizes the highly sensitive and rapid nature of this method, allowing the early detection of active toxins.https://www.mdpi.com/2072-6651/11/3/174ricinabrindigital holographic microscopymorphologyintoxicationholomonitor |
| spellingShingle | Efi Makdasi Orly Laskar Elad Milrot Ofir Schuster Shlomo Shmaya Shmuel Yitzhaki Whole-Cell Multiparameter Assay for Ricin and Abrin Activity-Based Digital Holographic Microscopy Toxins ricin abrin digital holographic microscopy morphology intoxication holomonitor |
| title | Whole-Cell Multiparameter Assay for Ricin and Abrin Activity-Based Digital Holographic Microscopy |
| title_full | Whole-Cell Multiparameter Assay for Ricin and Abrin Activity-Based Digital Holographic Microscopy |
| title_fullStr | Whole-Cell Multiparameter Assay for Ricin and Abrin Activity-Based Digital Holographic Microscopy |
| title_full_unstemmed | Whole-Cell Multiparameter Assay for Ricin and Abrin Activity-Based Digital Holographic Microscopy |
| title_short | Whole-Cell Multiparameter Assay for Ricin and Abrin Activity-Based Digital Holographic Microscopy |
| title_sort | whole cell multiparameter assay for ricin and abrin activity based digital holographic microscopy |
| topic | ricin abrin digital holographic microscopy morphology intoxication holomonitor |
| url | https://www.mdpi.com/2072-6651/11/3/174 |
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