scholarly journals A Portable Live-Cell Imaging System With an Invert-Upright-Convertible Architecture and a Mini-Bioreactor for Long-Term Simultaneous Cell Imaging, Chemical Sensing, and Electrophysiological Recording

IEEE Access ◽  
2018 ◽  
Vol 6 ◽  
pp. 11063-11075 ◽  
Author(s):  
Dhanesh Kattipparambil Rajan ◽  
Joose Kreutzer ◽  
Hannu Valimaki ◽  
Mari Pekkanen-Mattila ◽  
Antti Ahola ◽  
...  
Keyword(s):  
Chemical Sensing ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Imaging System ◽  
Live Cell ◽  
Electrophysiological Recording

scholarly journals A portable cell culture system with a simple microscope, oxygen sensor and MEA amplifier

2019 ◽  
Author(s):  
Dhanesh Kattipparambil Rajan
Keyword(s):  
Cell Culture ◽  
Chemical Sensing ◽  
Live Cell Imaging ◽  
Oxygen Sensor ◽  
Cell Imaging ◽  
Imaging System ◽  
Electrophysiological Recording

Cell culture in-vitro is a well-known method to develop cell and disease models for studying physiologically relevant mechanisms and responses for various applications in life sciences. Conventional methods for instance, using static culture flasks or well plates, have limitations, as these cannot provide accurate tractable models for advanced studies. However, microscale systems can overcome this since they mimic the cells' natural microenvironment adequately. We have developed a portable live-cell imaging system with an invert-upright-convertible architecture and a mini-bioreactor for long-term simultaneous cell imaging and analysis, chemical sensing and electrophysiological recording. Our system integrates biocompatible cell-friendly materials with modular measurement schemes and precise environment control and can be automated. High quality time-lapse cell imaging is hugely useful in cell/disease models. However, integration of advanced in-vitro systems into benchtop microscopes for in-situ imaging is tricky and challenging. This is especially true with device based biological systems, such as lab/organ/body-on-chips, or mini-bioreactors/microfluidic systems. They face issues ranging from optical and physical geometry incompatibilities to difficulties in connectivity of flow and perfusion systems. However, the novel modular system we have developed either as an inverted or as an upright system can easily accommodate virtually any in-vitro devices. Furthermore, it can accept additional sensor or measurement devices quite freely. Cell characterization, differentiation, chemical sensing, drug screening, microelectrode-array-electrophysiological recordings, and cell stimulation can be carried out with simultaneous in-situ imaging and analysis. Moreover, our system can be configured to capture images from regions that are otherwise inaccessible by conventional microscopes, for example, cells cultured on physical or biochemical sensor systems. We demonstrate the system for video-based beating analysis of cardiomyocytes, cell orientation analysis on nanocellulose, and simultaneous long-term in-situ microscopy with pO2 and temperature sensing. The compact microscope as such is comparable to standard phase-contrast-microscopes without any detectable aberrations and is useful practically for any in-situ microscopy demands. [1] D. K. Rajan et al., "A Portable Live-Cell Imaging System With an Invert-Upright-Convertible Architecture and a Mini-Bioreactorfor Long-Term Simultaneous Cell Imaging, Chemical Sensing, and Electrophysiological Recording," in IEEE Access, vol. 6,pp. 11063-11075, 2018. doi: 10.1109/ACCESS.2018.2804378. [2] Antti-Juhana Mäki et al., "A Portable Microscale Cell Culture System with Indirect Temperature Control"SLAS TECHNOLOGY: Translating Life Sciences Innovation 2018 23:6, 566-579. [3] Hannu Välimäki et al.,Fluorimetric oxygen sensor with an efficient optical read-out for in vitro cellmodels,Sensors and Actuators B: Chemical,Volume 249,2017,pp.738-746,ISSN 0925-4005,https://doi.org/10.1016/j.snb.2017.04.182.

scholarly journals A high-throughput screening assay based on automated microscopy for monitoring antibiotic susceptibility of Mycobacterium tuberculosis phenotypes

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Sadaf Kalsum ◽  
Blanka Andersson ◽  
Jyotirmoy Das ◽  
Thomas Schön ◽  
Maria Lerm
Keyword(s):  
Mycobacterium Tuberculosis ◽  
High Throughput ◽  
High Throughput Screening ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Imaging System ◽  
Aggregate Size ◽  
Live Cell ◽  
Screening Assay ◽  
High Throughput Screening Assay

Abstract Background Efficient high-throughput drug screening assays are necessary to enable the discovery of new anti-mycobacterial drugs. The purpose of our work was to develop and validate an assay based on live-cell imaging which can monitor the growth of two distinct phenotypes of Mycobacterium tuberculosis and to test their susceptibility to commonly used TB drugs. Results Both planktonic and cording phenotypes were successfully monitored as fluorescent objects using the live-cell imaging system IncuCyte S3, allowing collection of data describing distinct characteristics of aggregate size and growth. The quantification of changes in total area of aggregates was used to define IC50 and MIC values of selected TB drugs which revealed that the cording phenotype grew more rapidly and displayed a higher susceptibility to rifampicin. In checkerboard approach, testing pair-wise combinations of sub-inhibitory concentrations of drugs, rifampicin, linezolid and pretomanid demonstrated superior growth inhibition of cording phenotype. Conclusions Our results emphasize the efficiency of using automated live-cell imaging and its potential in high-throughput whole-cell screening to evaluate existing and search for novel antimycobacterial drugs.

scholarly journals A live-cell imaging system for visualizing the transport of Marburg virus nucleocapsid-like structures

2019 ◽  
Vol 16 (1) ◽  
Author(s):  
Yuki Takamatsu ◽  
Olga Dolnik ◽  
Takeshi Noda ◽  
Stephan Becker
Keyword(s):  
Live Cell Imaging ◽  
Intracellular Transport ◽  
Actin Polymerization ◽  
Ebola Virus ◽  
Cell Imaging ◽  
Temporal Dynamics ◽  
Imaging System ◽  
Marburg Virus ◽  
Live Cell ◽  
Infected Cells

Abstract Background Live-cell imaging is a powerful tool for visualization of the spatio-temporal dynamics of moving signals in living cells. Although this technique can be utilized to visualize nucleocapsid transport in Marburg virus (MARV)- or Ebola virus-infected cells, the experiments require biosafety level-4 (BSL-4) laboratories, which are restricted to trained and authorized individuals. Methods To overcome this limitation, we developed a live-cell imaging system to visualize MARV nucleocapsid-like structures using fluorescence-conjugated viral proteins, which can be conducted outside BSL-4 laboratories. Results Our experiments revealed that nucleocapsid-like structures have similar transport characteristics to those of nucleocapsids observed in MARV-infected cells, both of which are mediated by actin polymerization. Conclusions We developed a non-infectious live cell imaging system to visualize intracellular transport of MARV nucleocapsid-like structures. This system provides a safe platform to evaluate antiviral drugs that inhibit MARV nucleocapsid transport.

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