In vitro cytotoxicity evaluation of cadmium by label‐free holographic microscopy

Among all environmental pollutants, the toxic heavy metal cadmium is considered as a human carcinogen. Cadmium may induce cell death by apoptosis in various cell types, although the underlying mechanisms are still unclear. In this paper we show how a label‐free digital holography (DH)‐based technique is able to quantify the evolution of key biophysical parameters of cells during the exposure to cadmium for the first time. Murine embryonic fibroblasts NIH 3T3 are chosen here as cellular model for studying the cadmium effects. The results demonstrate that DH is able to retrieve the temporal evolution of different key parameters such as cell volume, projected area, cell thickness and dry mass, thus providing a full quantitative characterization of the cell physical behaviour during cadmium exposure. Our results show that the label‐free character of the technique would allow biologists to perform systematic and reliable studies on cell death process induced by cadmium and we believe that more in general this can be easily extended to others heavy metals, thus avoiding the time‐consuming, expensive and invasive label‐based procedures used nowadays in the field. In fact, pollution by heavy metals is severe issue that needs rapid and reliable methods to be settled.

[1]  J. Cidlowski,et al.  Analysis of glucocorticoid actions on rat thymocyte deoxyribonucleic acid by fluorescence-activated flow cytometry. , 1988, Endocrinology.

[2]  Elisabeth Heseltine,et al.  Meeting of the IARC working group on beryllium, cadmium, mercury and exposures in the glass manufacturing industry. , 1993, Scandinavian journal of work, environment & health.

[3]  H. Vainio,et al.  Meeting of the IARC working group on occupational exposures of hairdressers and barbers and on some colorings and dyestuffs. , 1993, Scandinavian journal of work, environment & health.

[4]  Y Ishizaki,et al.  Normotonic cell shrinkage because of disordered volume regulation is an early prerequisite to apoptosis. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[5]  H. Haase,et al.  Induction of Apoptosis in Mammalian Cells by Cadmium and Zinc , 2002 .

[6]  T. Cover,et al.  Recovery from lactacidosis‐induced glial cell swelling with the aid of exogenous anion channels , 2003, Glia.

[7]  Lars Jarup,et al.  Hazards of heavy metal contamination. , 2003 .

[8]  M. Brzóska,et al.  Antioxidant enzymes activity and lipid peroxidation in liver and kidney of rats exposed to cadmium and ethanol. , 2004, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[9]  E. Cuche,et al.  Measurement of the integral refractive index and dynamic cell morphometry of living cells with digital holographic microscopy. , 2005, Optics express.

[10]  A. Petrunkina,et al.  Regulatory and necrotic volume increase in boar spermatozoa , 2005, Journal of cellular physiology.

[11]  D. Groneberg,et al.  Journal of Occupational Medicine and Toxicology the Toxicity of Cadmium and Resulting Hazards for Human Health , 2006 .

[12]  John A Timbrell,et al.  In vitro cytotoxicity assays: comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride. , 2006, Toxicology letters.

[13]  C. Fang-Yen,et al.  Tomographic phase microscopy , 2008, Nature Methods.

[14]  Markus Fratz,et al.  Noninvasive time-dependent cytometry monitoring by digital holography. , 2007, Journal of biomedical optics.

[15]  Patrik Langehanenberg,et al.  Integral refractive index determination of living suspension cells by multifocus digital holographic phase contrast microscopy. , 2007, Journal of biomedical optics.

[16]  C. Ríos,et al.  Cadmium neurotoxicity. , 2007, Environmental toxicology and pharmacology.

[17]  Mark H. Ellisman,et al.  Correlated three-dimensional light and electron microscopy reveals transformation of mitochondria during apoptosis , 2007, Nature Cell Biology.

[18]  Adam Wax,et al.  Light scattering measurements of subcellular structure provide noninvasive early detection of chemotherapy-induced apoptosis. , 2009, Cancer research.

[19]  T. Cirillo,et al.  Survey of lead, cadmium, mercury and arsenic in seafood purchased in Campania, Italy , 2010, Food additives & contaminants. Part B, Surveillance.

[20]  Myung K. Kim,et al.  Applications of Digital Holography in Biomedical Microscopy , 2010 .

[21]  Zhuo Wang,et al.  Optical measurement of cycle-dependent cell growth , 2011, Proceedings of the National Academy of Sciences.

[22]  Patrik Langehanenberg,et al.  Differential cytotoxic actions of Shiga toxin 1 and Shiga toxin 2 on microvascular and macrovascular endothelial cells , 2010, Thrombosis and Haemostasis.

[23]  Christian Depeursinge,et al.  Early Cell Death Detection with Digital Holographic Microscopy , 2012, PloS one.

[24]  Alexander Khmaladze,et al.  Cell volume changes during apoptosis monitored in real time using digital holographic microscopy. , 2012, Journal of structural biology.

[25]  A. van Bokhoven,et al.  Cadmium Induces p53-Dependent Apoptosis in Human Prostate Epithelial Cells , 2012, PloS one.

[26]  P. Marquet,et al.  Marker-free phase nanoscopy , 2013, Nature Photonics.

[27]  Peter Vandenabeele,et al.  Determination of apoptotic and necrotic cell death in vitro and in vivo. , 2013, Methods.

[28]  P Memmolo,et al.  Digital holography as a method for 3D imaging and estimating the biovolume of motile cells. , 2013, Lab on a chip.

[29]  Jérôme Parent,et al.  Label-free cytotoxicity screening assay by digital holographic microscopy. , 2013, Assay and drug development technologies.

[30]  Myung K. Kim,et al.  Review of digital holographic microscopy for three-dimensional profiling and tracking , 2014 .

[31]  F. Flamigni,et al.  Cell death in human articular chondrocyte: a morpho-functional study in micromass model , 2014, Apoptosis.

[32]  Benjamin Rappaz,et al.  Digital Holographic Microscopy: A Quantitative Label-Free Microscopy Technique for Phenotypic Screening , 2014, Combinatorial chemistry & high throughput screening.

[33]  Steffi Ketelhut,et al.  Multimodal label-free in vitro toxicity testing with digital holographic microscopy , 2014, Photonics Europe.

[34]  Yu Sa,et al.  A quantitative method for measurement of HL-60 cell apoptosis based on diffraction imaging flow cytometry technique. , 2014, Biomedical optics express.

[35]  Steffi Ketelhut,et al.  Quantitative Stain-Free and Continuous Multimodal Monitoring of Wound Healing In Vitro with Digital Holographic Microscopy , 2014, PloS one.

[36]  Pasquale Memmolo,et al.  3D morphometry of red blood cells by digital holography , 2014, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[37]  Torsten Wittmann,et al.  Fluorescence live cell imaging. , 2014, Methods in cell biology.

[38]  M. Islam,et al.  Heavy metals in cereals and pulses: health implications in Bangladesh. , 2014, Journal of agricultural and food chemistry.

[39]  N. Kasai,et al.  Time-lapse imaging of morphological changes in a single neuron during the early stages of apoptosis using scanning ion conductance microscopy. , 2015, Journal of structural biology.

[40]  Vittorio Bianco,et al.  Diagnostic Tools for Lab-on-Chip Applications Based on Coherent Imaging Microscopy , 2015, Proceedings of the IEEE.

[41]  Chunye Lin,et al.  A review of soil cadmium contamination in China including a health risk assessment , 2015, Environmental Science and Pollution Research.

[42]  Y. Seo,et al.  An Overview of Carcinogenic Heavy Metal: Molecular Toxicity Mechanism and Prevention , 2015, Journal of cancer prevention.

[43]  Anna Mölder,et al.  Induction of morphological changes in death-induced cancer cells monitored by holographic microscopy. , 2015, Journal of structural biology.

[44]  Pasquale Memmolo,et al.  Recent advances in holographic 3D particle tracking , 2015 .

[45]  Björn Kemper,et al.  Virulence from vesicles: Novel mechanisms of host cell injury by Escherichia coli O104:H4 outbreak strain , 2015, Scientific Reports.

[46]  S. van de Linde,et al.  Light-induced cell damage in live-cell super-resolution microscopy , 2015, Scientific Reports.

[47]  Sarah Mues,et al.  Novel optical approaches for label-free quantification of nano-cytotoxic effects , 2016, SPIE BiOS.

[48]  A. Ozcan,et al.  High-Throughput Quantification of Nanoparticle Degradation Using Computational Microscopy and Its Application to Drug Delivery Nanocapsules , 2017 .

[49]  Pasquale Memmolo,et al.  Tomographic flow cytometry by digital holography , 2016, Light: Science & Applications.

[50]  Serge Monneret,et al.  Optical imaging and characterization of graphene and other 2D materials using quantitative phase microscopy , 2017 .

[51]  Björn Kemper,et al.  Quantitative phase imaging for cell culture quality control , 2017, Cytometry. Part A : the journal of the International Society for Analytical Cytology.