Miniaturized microscope for high throughput screening of tumor spheroids in microfluidic devices

2018 ◽  
Author(s):  
Carlos Ortiz de Solórzano ◽  
Javier Uranga ◽  
Alejandro Rodríguez-Pena ◽  
Desire Gahigiro
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Microfluidic Devices ◽  
Tumor Spheroids

Detection of Respiratory Viruses with Plastic High Throughput Screening Devices

2006 ◽  
Vol 950 ◽  
Author(s):  
Zhengshan Zhao ◽  
Gerardo A. Diaz-Quijada ◽  
Régis Peytavi ◽  
Éric LeBlanc ◽  
Johanne Frenette ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Low Cost ◽  
Genetic Material ◽  
Microfluidic Devices ◽  
Medical Diagnostics ◽  
Human Enterovirus ◽  
Plastic Substrate ◽  
Rt Pcr ◽  
Genomics And Proteomics

ABSTRACTMicroarrays have become one of the most convenient tools for high throughput screening and have catalyzed major advances in genomics and proteomics. Other important applications can be found in medical diagnostics, detection of biothreats, drug discovery, etc. Integration of microarrays with microfluidic devices can be highly advantageous in terms of portability, shorter analysis time and lower consumption of expensive biological analytes. Since fabrication of microfluidic devices using traditional materials such as glass is rather expensive, there is a high interest in employing polymeric materials as a low cost alternative suitable for mass production. We present proof-of-concept DNA arrays on a plastic platform for the detection of four important respiratory pathogens: Influenza A virus, respiratory syncytial virus, human enterovirus, and human metapneumovirus.This was accomplished by amplifying the genetic material from the viruses and simultaneously labeling the amplicons with a fluorescent dye (Cy3) via a highly sensitive multiplex Reverse Transcription Polymerase Chain Reaction (RT-PCR). The resultant RT-PCR product was hybridized, without further purification, with an array of specific oligonucleotide probes (20 mers) that had been covalently bound to a plastic substrate. Results indicate a high signal to background ratio that is comparable to commercially available microarray glass slides. In addition, 5 minute hybridization on this plastic substrate has been demonstrated using a centrifugal microfluidic platform, paving the way to a rapid medical diagnostic device for point-of-care use that is based on a low-cost portable Micro-Total-Analysis-System (μ-TAS).

Toward High-Throughput Screening of NAD(P)-Dependent Oxidoreductases Using Boron-Doped Diamond Microelectrodes and Microfluidic Devices

2014 ◽  
Vol 86 (19) ◽  
pp. 9570-9575 ◽  
Author(s):  
Ryo Oyobiki ◽  
Taisuke Kato ◽  
Michinobu Katayama ◽  
Ai Sugitani ◽  
Takeshi Watanabe ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Microfluidic Devices ◽  
Boron Doped Diamond ◽  
Boron Doped

scholarly journals Microfluidic devices for cell based high throughput screening

Lab on a Chip ◽  
2010 ◽  
Vol 10 (3) ◽  
pp. 341-348 ◽  
Author(s):  
Sarvesh Upadhyaya ◽  
P. Ravi Selvaganapathy
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Microfluidic Devices

scholarly journals RNAi High-Throughput Screening of Single- and Multi-Cell-Type Tumor Spheroids: A Comprehensive Analysis in Two and Three Dimensions

2017 ◽  
Vol 22 (5) ◽  
pp. 525-536 ◽  
Author(s):  
Jiaqi Fu ◽  
Daniel Fernandez ◽  
Marc Ferrer ◽  
Steven A. Titus ◽  
Eugen Buehler ◽  
...  
Keyword(s):  
Drug Discovery ◽  
High Throughput ◽  
High Throughput Screening ◽  
Attrition Rate ◽  
Three Dimensions ◽  
Cell Type ◽  
Tumor Spheroids ◽  
Flat Bottom ◽  
Cancer Subtypes

The widespread use of two-dimensional (2D) monolayer cultures for high-throughput screening (HTS) to identify targets in drug discovery has led to attrition in the number of drug targets being validated. Solid tumors are complex, aberrantly growing microenvironments that harness structural components from stroma, nutrients fed through vasculature, and immunosuppressive factors. Increasing evidence of stromally-derived signaling broadens the complexity of our understanding of the tumor microenvironment while stressing the importance of developing better models that reflect these interactions. Three-dimensional (3D) models may be more sensitive to certain gene-silencing events than 2D models because of their components of hypoxia, nutrient gradients, and increased dependence on cell-cell interactions and therefore are more representative of in vivo interactions. Colorectal cancer (CRC) and breast cancer (BC) models composed of epithelial cells only, deemed single-cell-type tumor spheroids (SCTS) and multi-cell-type tumor spheroids (MCTS), containing fibroblasts were developed for RNAi HTS in 384-well microplates with flat-bottom wells for 2D screening and round-bottom, ultra-low-attachment wells for 3D screening. We describe the development of a high-throughput assay platform that can assess physiologically relevant phenotypic differences between screening 2D versus 3D SCTS, 3D SCTS, and MCTS in the context of different cancer subtypes. This assay platform represents a paradigm shift in how we approach drug discovery that can reduce the attrition rate of drugs that enter the clinic.

scholarly journals 3D hanging spheroid plate for high-throughput CAR T cell cytotoxicity assay

2022 ◽  
Vol 20 (1) ◽  
Author(s):  
Zhenzhong Chen ◽  
Seokgyu Han ◽  
Arleen Sanny ◽  
Dorothy Leung-Kwan Chan ◽  
Danny van Noort ◽  
...  
Keyword(s):  
T Cells ◽  
Tumor Cells ◽  
High Throughput ◽  
High Throughput Screening ◽  
Tumor Type ◽  
Tumor Spheroids ◽  
Cytotoxic Effects ◽  
Cytotoxicity Assays ◽  
Car T Cells ◽  
Car T

Abstract Background Most high-throughput screening (HTS) systems studying the cytotoxic effect of chimeric antigen receptor (CAR) T cells on tumor cells rely on two-dimensional cell culture that does not recapitulate the tumor microenvironment (TME). Tumor spheroids, however, can recapitulate the TME and have been used for cytotoxicity assays of CAR T cells. But a major obstacle to the use of tumor spheroids for cytotoxicity assays is the difficulty in separating unbound CAR T and dead tumor cells from spheroids. Here, we present a three-dimensional hanging spheroid plate (3DHSP), which facilitates the formation of spheroids and the separation of unbound and dead cells from spheroids during cytotoxicity assays. Results The 3DHSP is a 24-well plate, with each well composed of a hanging dripper, spheroid wells, and waste wells. In the dripper, a tumor spheroid was formed and mixed with CAR T cells. In the 3DHSP, droplets containing the spheroids were deposited into the spheroid separation well, where unbound and dead T and tumor cells were separated from the spheroid through a gap into the waste well by tilting the 3DHSP by more than 20°. Human epidermal growth factor receptor 2 (HER2)-positive tumor cells (BT474 and SKOV3) formed spheroids of approximately 300–350 μm in diameter after 2 days in the 3DHSP. The cytotoxic effects of T cells engineered to express CAR recognizing HER2 (HER2-CAR T cells) on these spheroids were directly measured by optical imaging, without the use of live/dead fluorescent staining of the cells. Our results suggest that the 3DHSP could be incorporated into a HTS system to screen for CARs that enable T cells to kill spheroids formed from a specific tumor type with high efficacy or for spheroids consisting of tumor types that can be killed efficiently by T cells bearing a specific CAR. Conclusions The results suggest that the 3DHSP could be incorporated into a HTS system for the cytotoxic effects of CAR T cells on tumor spheroids. Graphical Abstract

Towards Low Cost Disposable High Throughput Screening Devices

2005 ◽  
Vol 897 ◽  
Author(s):  
Gerardo Antonio Diaz-Quijada ◽  
Regis Peytavi ◽  
André Nantel ◽  
Emmanuel Roy ◽  
Michel G. Bergeron ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Low Cost ◽  
Polymeric Materials ◽  
Microfluidic Devices ◽  
Medical Diagnostics ◽  
Covalent Immobilization ◽  
Plastic Materials ◽  
Genomics And Proteomics ◽  
Biological Analytes

AbstractMicroarrays have become one of the most convenient tools for high throughput screening, supporting major advances in genomics and proteomics. Other important applications can be found in medical diagnostics, detection of biothreats, drug discovery, etc. Integration of microarrays with microfluidic devices can be highly advantageous in terms of portability, shorter analysis time and lower consumption of expensive biological analytes. Since fabrication of microfluidic devices using traditional materials such as glass is rather expensive, there is a high interest in employing polymeric materials as a low cost alternative that is suitable for mass production. A number of commercially available plastic materials were reviewed for this purpose and poly(methylmethacrylate) and Zeonor™ 1060R were identified as promising candidates, for which methods for surface modification and covalent immobilization of DNA oligonucleotide were developed. In addition, we present proof-of-concept plastic-based microarrays with and without integration with microfluidics.

Microfabrication of single-use plastic microfluidic devices for high-throughput screening and DNA analysis

2002 ◽  
Vol 7 (5-6) ◽  
pp. 265-268 ◽  
Author(s):  
A. Gerlach ◽  
G. Knebel ◽  
A.E. Guber ◽  
M. Heckele ◽  
D. Herrmann ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Dna Analysis ◽  
Microfluidic Devices ◽  
Single Use

scholarly journals 3D Hanging Spheroid Plate for High-Throughput CAR T Cell Cytotoxicity Assay

2021 ◽  
Author(s):  
Zhenzhong Chen ◽  
Seokgyu Han ◽  
Arleen Sanny ◽  
Dorothy Leung-Kwan Chan ◽  
Danny van Noort ◽  
...  
Keyword(s):  
T Cells ◽  
Tumor Cells ◽  
High Throughput ◽  
High Throughput Screening ◽  
Tumor Type ◽  
Tumor Spheroids ◽  
Cytotoxic Effects ◽  
Cytotoxicity Assays ◽  
Car T Cells ◽  
Car T

Abstract Background Most high-throughput screening (HTS) systems for the cytotoxic effect of chimeric antigen receptor (CAR) T cells on tumor cells rely on two-dimensional cell culture that does not suitably recapitulate the tumor microenvironment (TME). Tumor spheroids can recapitulate TME and have been used for cytotoxicity assays of CAR T cells. However, a major obstacle to the use of tumor spheroids for cytotoxicity assays is the difficulty in separating unbound CAR T and dead tumor cells from spheroids. Here, we present a three-dimensional hanging spheroid plate (3DHSP), which facilitates the formation of spheroids and the separation of unbound and dead cells from spheroids during cytotoxicity assays. Results The 3DHSP is a 24-well plate, with each well composed of a hanging dripper, spheroid wells, and waste wells. In the dripper, a tumor spheroid was formed and mixed with CAR T cells. In 3DHSP, droplets containing the spheroids were deposited into the spheroid separation well, where unbound and dead T and tumor cells were separated from the spheroid through a gap into the waste well by tilting the 3DHSP by more than 20°. Human epidermal growth factor receptor 2 (HER2)-positive tumor cells (BT474 and SKOV3) formed spheroids of approximately 300–350 mm in diameter after 2 d in the 3DHSP. The cytotoxic effects of T cells engineered to express CAR recognizing HER2 (HER2-CAR T cells) on these spheroids were directly measured by optical imaging, without the use of live and dead fluorescent staining of the cells. Our results suggest that the 3DHSP could be incorporated into an HTS system to screen for CARs that enable T cells to kill spheroids formed from a specific tumor type with high efficacy or for spheroids consisting of tumor types that can be killed efficiently by T cells bearing a specific CAR. Conclusions The results suggest that the 3DHSP could be incorporated into an HTS system for the cytotoxic effects of CAR T cells on tumor spheroids.

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