The Large‐Scale Digital Cell Analysis System: an open system for nonperturbing live cell imaging

The Large‐Scale Digital Cell Analysis System (LSDCAS) was designed to provide a highly extensible open source live cell imaging system. Analysis of cell growth data has demonstrated a lack of perturbation in cells imaged using LSDCAS, through reference to cell growth data from cells growing in CO2 incubators. LSDCAS consists of data acquisition, data management and data analysis software, and is currently a Core research facility at the Holden Comprehensive Cancer Center at the University of Iowa. Using LSDCAS analysis software, this report and others show that although phase‐contrast imaging has no apparent effect on cell growth kinetics and viability, fluorescent image acquisition in the cell lines tested caused a measurable level of growth perturbation using LSDCAS. This report describes the current design of the system, reasons for the implemented design, and details its basic functionality. The LSDCAS software runs on the GNU/Linux operating system, and provides easy to use, graphical programs for data acquisition and quantitative analysis of cells imaged with phase‐contrast or fluorescence microscopy (alone or in combination), and complete source code is freely available under the terms of the GNU Public Software License at the project website (http://lsdcas.engineering.uiowa.edu).

[1]  Yi Li,et al.  Ameboid cell motility: a model and inverse problem, with an application to live cell imaging data. , 2007, Journal of theoretical biology.

[2]  E Pomplun,et al.  Ratio of complex double strand break damage induced by 125IUdR and 123IUdR correlates with experimental in vitro cell killing effectiveness. , 2002, Radiation protection dosimetry.

[3]  Fiorenza Ianzini,et al.  The Large Scale Digital Cell Analysis System and its use in the quantitative analysis of cell populations , 2002, 2nd Annual International IEEE-EMBS Special Topic Conference on Microtechnologies in Medicine and Biology. Proceedings (Cat. No.02EX578).

[4]  F Ianzini,et al.  Spontaneous premature chromosome condensation and mitotic catastrophe following irradiation of HeLa S3 cells. , 1997, International journal of radiation biology.

[5]  Fiorenza Ianzini,et al.  Lack of p53 function promotes radiation-induced mitotic catastrophe in mouse embryonic fibroblast cells , 2006, Cancer Cell International.

[6]  Allen W Cowley.  The elusive field of systems biology. , 2004, Physiological genomics.

[7]  Rainer Duden,et al.  Live cell imaging: the 'green revolution' continues apace. , 2002, Trends in cell biology.

[8]  Milan Sonka,et al.  Mitotic cell recognition with hidden Markov models , 2004, Medical Imaging: Image-Guided Procedures.

[9]  Ram Dixit,et al.  Cell damage and reactive oxygen species production induced by fluorescence microscopy: effect on mitosis and guidelines for non-invasive fluorescence microscopy. , 2003, The Plant journal : for cell and molecular biology.

[10]  J. Price,et al.  Comparison of phase-contrast and fluorescence digital autofocus for scanning microscopy. , 1994, Cytometry.

[11]  Tokuko Haraguchi,et al.  Live cell imaging: approaches for studying protein dynamics in living cells. , 2002, Cell structure and function.

[12]  Amane Shiohara,et al.  On the Cyto‐Toxicity Caused by Quantum Dots , 2004, Microbiology and immunology.

[13]  Victoria J Allan,et al.  Light Microscopy Techniques for Live Cell Imaging , 2003, Science.

[14]  M. L. Pepper,et al.  Computer-assisted analysis of time-lapse cinemicrographs of cultured cells. , 1978, Computers and biomedical research, an international journal.

[15]  Timothy A. Skimina,et al.  Activation of flavin-containing oxidases underlies light-induced production of H2O2 in mammalian cells. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[16]  H. Netten,et al.  Autofocusing in microscopy based on the OTF and sampling , 1994 .

[17]  F Ianzini,et al.  Development of the large scale digital cell analysis system. , 2002, Radiation protection dosimetry.

[18]  Milan Sonka,et al.  Cell Segmentation, Tracking, and Mitosis Detection Using Temporal Context , 2005, MICCAI.

[19]  Hassan S. Bazzi,et al.  Differences in subcellular distribution and toxicity of green and red emitting CdTe quantum dots , 2005, Journal of Molecular Medicine.

[20]  H. Kitano.  Systems Biology: A Brief Overview , 2002, Science.

[21]  D. Vollath.  Automatic focusing by correlative methods , 1987 .

[22]  D. A. Lee,et al.  Live cell imaging using confocal microscopy induces intracellular calcium transients and cell death. , 2003, American journal of physiology. Cell physiology.

[23]  F Ianzini,et al.  Delayed DNA damage associated with mitotic catastrophe following X-irradiation of HeLa S3 cells. , 1998, Mutagenesis.

[24]  W Böcker,et al.  A fast autofocus unit for fluorescence microscopy. , 1997, Physics in medicine and biology.

[25]  Milan Sonka,et al.  Segmentation and quantitative analysis of the living tumor cells using large-scale digital cell analysis system , 2004, SPIE Medical Imaging.