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Whole-Cell Multiparameter Assay for Ricin and Abrin Activity-Based Digital Holographic Microscopy

The Department of Infectious Diseases, Israel Institute for Biological Research, Ness-Ziona 74100, Israel
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Toxins 2019, 11(3), 174; https://doi.org/10.3390/toxins11030174
Received: 6 February 2019 / Revised: 11 March 2019 / Accepted: 15 March 2019 / Published: 22 March 2019
(This article belongs to the Collection Toxic and Pharmacological Effect of Plant Toxins)
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Abstract

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. View Full-Text
Keywords: ricin; abrin; digital holographic microscopy; morphology; intoxication; holomonitor ricin; abrin; digital holographic microscopy; morphology; intoxication; holomonitor
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Makdasi, E.; Laskar, O.; Milrot, E.; Schuster, O.; Shmaya, S.; Yitzhaki, S. Whole-Cell Multiparameter Assay for Ricin and Abrin Activity-Based Digital Holographic Microscopy. Toxins 2019, 11, 174.

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