Real-time quantitative fluorescence measurement of microscale cell culture analog systems

2007 ◽  
Author(s):  
Taek-il Oh ◽  
Donghyun Kim ◽  
Daniel Tatosian ◽  
Jong Hwan Sung ◽  
Michael Shuler
Keyword(s):  
Cell Culture ◽  
Real Time ◽  
Fluorescence Measurement ◽  
Analog Systems ◽  
Quantitative Fluorescence

scholarly journals Real-time monitoring of liver fibrosis through embedded sensors in a microphysiological system

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Hafiz Muhammad Umer Farooqi ◽  
Bohye Kang ◽  
Muhammad Asad Ullah Khalid ◽  
Abdul Rahim Chethikkattuveli Salih ◽  
Kinam Hyun ◽  
...  
Keyword(s):  
Cell Culture ◽  
Liver Fibrosis ◽  
Real Time ◽  
Hepatic Fibrosis ◽  
Transforming Growth Factor ◽  
Cell Monolayer ◽  
Embedded Sensors ◽  
Matrix Deposition ◽  
Tgf Β1

AbstractHepatic fibrosis is a foreshadowing of future adverse events like liver cirrhosis, liver failure, and cancer. Hepatic stellate cell activation is the main event of liver fibrosis, which results in excessive extracellular matrix deposition and hepatic parenchyma's disintegration. Several biochemical and molecular assays have been introduced for in vitro study of the hepatic fibrosis progression. However, they do not forecast real-time events happening to the in vitro models. Trans-epithelial electrical resistance (TEER) is used in cell culture science to measure cell monolayer barrier integrity. Herein, we explored TEER measurement's utility for monitoring fibrosis development in a dynamic cell culture microphysiological system. Immortal HepG2 cells and fibroblasts were co-cultured, and transforming growth factor β1 (TGF-β1) was used as a fibrosis stimulus to create a liver fibrosis-on-chip model. A glass chip-based embedded TEER and reactive oxygen species (ROS) sensors were employed to gauge the effect of TGF-β1 within the microphysiological system, which promotes a positive feedback response in fibrosis development. Furthermore, albumin, Urea, CYP450 measurements, and immunofluorescent microscopy were performed to correlate the following data with embedded sensors responses. We found that chip embedded electrochemical sensors could be used as a potential substitute for conventional end-point assays for studying fibrosis in microphysiological systems.

scholarly journals Real-time imaging of cellular forces using optical interference

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Andrew T. Meek ◽  
Nils M. Kronenberg ◽  
Andrew Morton ◽  
Philipp Liehm ◽  
Jan Murawski ◽  
...  
Keyword(s):  
Cell Culture ◽  
Real Time ◽  
High Throughput ◽  
Mechanical Activity ◽  
Dynamic Processes ◽  
Imaging Method ◽  
Neonatal Cardiomyocytes ◽  
A Cell ◽  
Cellular Forces ◽  
Time Acquisition

AbstractImportant dynamic processes in mechanobiology remain elusive due to a lack of tools to image the small cellular forces at play with sufficient speed and throughput. Here, we introduce a fast, interference-based force imaging method that uses the illumination of an elastic deformable microcavity with two rapidly alternating wavelengths to map forces. We show real-time acquisition and processing of data, obtain images of mechanical activity while scanning across a cell culture, and investigate sub-second fluctuations of the piconewton forces exerted by macrophage podosomes. We also demonstrate force imaging of beating neonatal cardiomyocytes at 100 fps which reveals mechanical aspects of spontaneous oscillatory contraction waves in between the main contraction cycles. These examples illustrate the wider potential of our technique for monitoring cellular forces with high throughput and excellent temporal resolution.

scholarly journals Detection of enterovirus in mussels from Morocco by cell culture and real-time PCR

2017 ◽  
Vol 16 (34) ◽  
pp. 1791-1799 ◽  
Author(s):  
Meryem Idrissi Azzouzi Lalla ◽  
Senouci Samira ◽  
El Qazoui Maria ◽  
Oumzil Hicham ◽  
Naciri Mariam
Keyword(s):  
Cell Culture ◽  
Real Time ◽  
Real Time Pcr

Detection and Quantification of Human Adenoviruses in Surface Waters by Nested PCR, TaqMan Real-Time PCR and Cell Culture Assays

2007 ◽  
Vol 191 (1-4) ◽  
pp. 83-93 ◽  
Author(s):  
M. Muscillo ◽  
M. Pourshaban ◽  
M. Iaconelli ◽  
S. Fontana ◽  
A. Di Grazia ◽  
...  
Keyword(s):  
Cell Culture ◽  
Real Time ◽  
Surface Waters ◽  
Real Time Pcr ◽  
Nested Pcr ◽  
Human Adenoviruses ◽  
Detection And Quantification

scholarly journals Recent Advances in Application of Biosensors in Tissue Engineering

2014 ◽  
Vol 2014 ◽  
pp. 1-18 ◽  
Author(s):  
Anwarul Hasan ◽  
Md Nurunnabi ◽  
Mahboob Morshed ◽  
Arghya Paul ◽  
Alessandro Polini ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Cell Culture ◽  
Small Molecules ◽  
Real Time ◽  
Inflammatory Cytokines ◽  
Serum Proteins ◽  
Molecular Size ◽  
Medical Diagnostics ◽  
Alpha Fetoprotein ◽  
Pharmaceutical Industries

Biosensors research is a fast growing field in which tens of thousands of papers have been published over the years, and the industry is now worth billions of dollars. The biosensor products have found their applications in numerous industries including food and beverages, agricultural, environmental, medical diagnostics, and pharmaceutical industries and many more. Even though numerous biosensors have been developed for detection of proteins, peptides, enzymes, and numerous other biomolecules for diverse applications, their applications in tissue engineering have remained limited. In recent years, there has been a growing interest in application of novel biosensors in cell culture and tissue engineering, for example, real-time detection of small molecules such as glucose, lactose, and H2O2as well as serum proteins of large molecular size, such as albumin and alpha-fetoprotein, and inflammatory cytokines, such as IFN-g and TNF-α. In this review, we provide an overview of the recent advancements in biosensors for tissue engineering applications.

scholarly journals A Modular Toolkit to Fabricate Microfluidic Devices for 3D Cell Culture and Near Real-time Measurements

2020 ◽  
Author(s):  
Giraso Kabandana ◽  
Adam Michael Ratajczak ◽  
Chengpeng Chen
Keyword(s):  
Cell Culture ◽  
Real Time ◽  
Low Cost ◽  
New Technology ◽  
Microfluidic Channel ◽  
3D Cell Culture ◽  
Microfluidic Devices ◽  
In Vitro Cell Cultures ◽  
Scoring Tools

Microfluidic technology has tremendously facilitated the development of in vitro cell cultures and studies. Conventionally, microfluidic devices are fabricated with extensive facilities by well-trained researchers, which hinders the widespread adoption of the technology for broader applications. Enlightened by the fact that low-cost microbore tubing is a natural microfluidic channel, we developed a series of adaptors in a toolkit that can twine, connect, organize, and configure the tubing to produce functional microfluidic units. Three subsets of the toolkit were thoroughly developed: the tubing and scoring tools, the flow adaptors, and the 3D cell culture suite. To demonstrate the usefulness and versatility of the toolkit, we assembled a microfluidic device and successfully applied it for 3D macrophage cultures, flow-based stimulation, and automated near real-time quantitation with new knowledge generated. Overall, we present a new technology that allows simple, fast, and robust assembly of customizable and scalable microfluidic devices with minimal facilities, which is broadly applicable to research that needs or could be enhanced by microfluidics.

scholarly journals Study Cellular Hypoxic Response under Cyclic Oxygen Gradients Generated in Microfluidic Devices Using Real-Time Fluorescence Imaging

2021 ◽  
Author(s):  
Dao-Ming Chang ◽  
Yi-Chung Tung
Keyword(s):  
Cell Culture ◽  
Intracellular Calcium ◽  
Real Time ◽  
Oxygen Tension ◽  
Calcium Concentration ◽  
A549 Cells ◽  
Cell Types ◽  
Hypoxic Response ◽  
Oxygen Gradient ◽  
Oxygen Gradients

Abstract In this paper, an integrated approach to study cellular hypoxic response under cyclic oxygen gradients is developed. In the experiments, a cell culture system based on a microfluidic device is constructed to generate cyclic oxygen gradients with fast response. The oxygen gradient is generated by alternatively introducing gases with specific compositions by a computer-controlled gas supply system into the microfluidic channels next to the cell culture one separated by thin channel walls. Observation of cellular hypoxic responses is performed using real-time fluorescence imaging of dyes sensitive to extra-and intra-cellular oxygen tensions as well as intracellular calcium concentrations. The capability of consistent cyclic oxygen gradient generation is confirmed by characterizing spatiotemporal oxygen tension profiles within the device. In the cell experiments, cellular hypoxic responses of human aortic smooth muscle cells (AoSMCs) and lung carcinoma epithelium (A549) cells including intracellular oxygen and calcium levels are measured. The relationships between the oxygen probe intensity and extracellular oxygen tension for both cell types are highly linear, and difference between the intensity measured from the two cell types is observed between 4 to 8% O2 culture environment. In addition, both cell types show detectable intracellular calcium concentration variation when the environmental O2 varies between normoxia and lower than 4%. The AoSMCs and A549 cells show decrease and increase of the intracellular calcium concentration during hypoxic response when facing change from normoxia to hypoxia environments, respectively. With the capability of real-time cellular responses monitoring under cyclic oxygen gradients, the developed approach provides a useful scheme to investigate cellular hypoxic response in vitro under microenvironments mimicking various in vivo physiological and pathological conditions.

scholarly journals Identification and evaluation of an appropriate housekeeping gene for real-time gene profiling of hepatocellular carcinoma cells cultured in three-dimensional scaffold

2021 ◽  
Author(s):  
Anjana Kaveri Badekila ◽  
Praveen Rai ◽  
Sudarshan Kini
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Culture ◽  
Real Time ◽  
Internal Control ◽  
18S Rrna ◽  
Three Dimensional ◽  
Melting Curve ◽  
3D Culture ◽  
Gene Profiling ◽  
3D Microenvironment

Abstract Assessing an optimal reference gene as an internal control for target gene normalization is important during quantitative real time polymerase chain reaction (RT-qPCR) of three-dimensional cell culture. Especially, gene profiling of cancer cells under a complex 3D microenvironment in a polymer scaffold provides a deeper understanding of recapitulation of in vivo tumors. In this aspect, expression of six housekeeping genes (HKG’s): glyceraldehyde-3-phosphodehydrogenase (GAPDH), β-actin (ACTB), beta-2-microglobulin (B2M), 18S ribosomal RNA (18S rRNA), peptidyl-propyl-isomerase A (PPIA), and ribosomal protein L13 (RPL-13)) during the monolayer culture (two-dimensional), and alginate-carboxymethylcellulose scaffold based three-dimensional (3D) cell culture conditioned up to 21 days was analyzed for hepatocellular carcinoma (Huh-7) cell line. The real-time gene expression using RT-qPCR of HCC spheroids in 3D culture were analyzed by determining the primer efficiency, melting curve and quantification cycle analysis of the selected candidate HKG’s. Further, RT-qPCR data were validated using analysis softwares i.e., geNorm and NormFinder for statistical significance. The study indicated RPL-13, 18S rRNA and B2M to be stable among selected referral HKG candidates and considered them as potential internal controls during varying cell culture conditions.

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