scholarly journals An inexpensive, customizable microscopy system for the automated quantification and characterization of multiple adherent cell types

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e4937 ◽  
Author(s):  
Vishwaratn Asthana ◽  
Yuqi Tang ◽  
Adam Ferguson ◽  
Pallavi Bugga ◽  
Anantratn Asthana ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Dynamic Range ◽  
Low Cost ◽  
Cell Types ◽  
Cell Counting ◽  
Adherent Cell ◽  
Essential Components ◽  
Automated Quantification ◽  
Automated Microscopy ◽  
Multiple Cell

Cell quantification assays are essential components of most biological and clinical labs. However, many currently available quantification assays, including flow cytometry and commercial cell counting systems, suffer from unique drawbacks that limit their overall efficacy. In order to address the shortcomings of traditional quantification assays, we have designed a robust, low-cost, automated microscopy-based cytometer that quantifies individual cells in a multiwell plate using tools readily available in most labs. Plating and subsequent quantification of various dilution series using the automated microscopy-based cytometer demonstrates the single-cell sensitivity, near-perfect R2 accuracy, and greater than 5-log dynamic range of our system. Further, the microscopy-based cytometer is capable of obtaining absolute counts of multiple cell types in one well as part of a co-culture setup. To demonstrate this ability, we recreated an experiment that assesses the tumoricidal properties of primed macrophages on co-cultured tumor cells as a proof-of-principle test. The results of the experiment reveal that primed macrophages display enhanced cytotoxicity toward tumor cells while simultaneously losing the ability to proliferate, an example of a dynamic interplay between two cell populations that our microscopy-based cytometer is successfully able to elucidate.

scholarly journals A customizable microscopy system for the automated quantification and characterization of multiple adherent cell types: an alternative to flow cytometry

2018 ◽  
Author(s):  
Vishwaratn Asthana ◽  
Yuqi Tang ◽  
Adam Ferguson ◽  
Pallavi Bugga ◽  
Anantratn Asthana ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Tumor Cells ◽  
Dynamic Range ◽  
Low Cost ◽  
Cell Types ◽  
Cell Counting ◽  
Adherent Cell ◽  
Optical Cell ◽  
Essential Components ◽  
Multiple Cell

Cell quantification assays are essential components of most biological and clinical labs. However, many currently available quantification assays, including flow cytometry and commercial cell counting systems, suffer from unique drawbacks that limit their overall efficacy. In order to address the shortcomings of traditional quantification assays, we have designed a robust, low-cost, automated optical cell cytometer that quantifies individual cells in a multiwell plate using tools readily available in most labs. Plating and subsequent quantification of various dilution series using the automated optical cytometer demonstrates the single-cell sensitivity, near-perfect R2 accuracy, and greater than 5-log dynamic range of our system. Further, the optical cytometer is capable of obtaining absolute counts of multiple cell types in one well as part of a co-culture setup. To demonstrate this ability, we recreated an experiment that assesses the tumoricidal properties of primed macrophages on co-cultured tumor cells as a proof-of-principle test. The results of the experiment reveal that primed macrophages display enhanced cytotoxicity towards tumor cells while simultaneously losing the ability to proliferate, an example of a dynamic interplay between two cell populations that our optical cytometer is successfully able to elucidate.

scholarly journals A customizable microscopy system for the automated quantification and characterization of multiple adherent cell types: an alternative to flow cytometry

2018 ◽  
Author(s):  
Vishwaratn Asthana ◽  
Yuqi Tang ◽  
Adam Ferguson ◽  
Pallavi Bugga ◽  
Anantratn Asthana ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Tumor Cells ◽  
Dynamic Range ◽  
Low Cost ◽  
Cell Types ◽  
Cell Counting ◽  
Adherent Cell ◽  
Optical Cell ◽  
Essential Components ◽  
Multiple Cell

Cell quantification assays are essential components of most biological and clinical labs. However, many currently available quantification assays, including flow cytometry and commercial cell counting systems, suffer from unique drawbacks that limit their overall efficacy. In order to address the shortcomings of traditional quantification assays, we have designed a robust, low-cost, automated optical cell cytometer that quantifies individual cells in a multiwell plate using tools readily available in most labs. Plating and subsequent quantification of various dilution series using the automated optical cytometer demonstrates the single-cell sensitivity, near-perfect R2 accuracy, and greater than 5-log dynamic range of our system. Further, the optical cytometer is capable of obtaining absolute counts of multiple cell types in one well as part of a co-culture setup. To demonstrate this ability, we recreated an experiment that assesses the tumoricidal properties of primed macrophages on co-cultured tumor cells as a proof-of-principle test. The results of the experiment reveal that primed macrophages display enhanced cytotoxicity towards tumor cells while simultaneously losing the ability to proliferate, an example of a dynamic interplay between two cell populations that our optical cytometer is successfully able to elucidate.

scholarly journals Label-free counting of affinity-enriched circulating tumor cells (CTCs) using a thermoplastic micro-Coulter counter (μCC)

The Analyst ◽  
2020 ◽  
Vol 145 (5) ◽  
pp. 1677-1686
Author(s):  
Cong Kong ◽  
Mengjia Hu ◽  
Kumuditha M. Weerakoon-Ratnayake ◽  
Malgorzata A. Witek ◽  
Kavya Dathathreya ◽  
...  
Keyword(s):  
Tumor Cell ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Low Cost ◽  
Circulating Tumor Cell ◽  
Free Cell ◽  
Coulter Counter ◽  
Fabrication Process ◽  
Cell Counting ◽  
Label Free

A micro-Coulter Counter (μCC) with simple and low cost fabrication process was developed and integrated with a circulating tumor cell (CTC) selection chip, providing label-free cell counting for CTC analysis.

scholarly journals The MEMIC: An ex vivo system to model the complexity of the tumor microenvironment

2021 ◽  
Author(s):  
Libuše Janská ◽  
Libi Anandi ◽  
Nell C. Kirchberger ◽  
Zoran S. Marinkovic ◽  
Logan T. Schachtner ◽  
...  
Keyword(s):  
Tumor Microenvironment ◽  
Tumor Cells ◽  
Ex Vivo ◽  
Tumor Stroma ◽  
Cell Types ◽  
Blood Stream ◽  
Direct Access ◽  
Heterogeneous Environments ◽  
Multiple Cell ◽  
Cost Efficient

ABSTRACTThere is an urgent need for accurate, scalable, and cost-efficient models of the complexity and heterogeneity of the tumor microenvironment. Here, we detail how to fabricate and use the Metabolic Microenvironment Chamber (MEMIC) – a 3D-printed ex vivo model of intratumoral heterogeneity. A major driver of the cellular and molecular diversity in tumors is the accessibility to the blood stream that provides key resources such as oxygen and nutrients. While some tumor cells have direct access to these resources, many others must survive under progressively more ischemic environments as they reside further from the vasculature. The MEMIC is designed to simulate the differential access to nutrients and allows co-culturing different cell types, such as tumor and immune cells. This system is optimized for live imaging and other microscopy-based approaches and it is a powerful tool to study tumor features such as the effect of nutrient scarcity on tumor-stroma interactions. Due to its adaptable design and full experimental control, the MEMIC can provide novel insights into the tumor microenvironment that would be difficult to obtain via other methods. As a proof of principle, we show that cells can sense gradual changes in metabolite concentration, and tune intracellular cell signaling to form multicellular spatial patterns of cell proliferation. We also show that ischemic macrophages reduce epithelial features in neighboring tumor cells highlighting the power of this system to study cell-cell interactions and non-cell autonomous effects of the metabolic microenvironment. We propose that the MEMIC can be easily adapted to study early development, ischemic stroke, and other systems where multiple cell types interact within heterogeneous environments.

scholarly journals Bidirectional and Opposite Effects of Naïve Mesenchymal Stem Cells Ontumor Growth and Progression

2019 ◽  
Vol 9 (4) ◽  
pp. 539-558 ◽  
Author(s):  
Faramarz Rahmatizadeh ◽  
Shiva Gholizadeh-Ghaleh Aziz ◽  
Khodadad Khodadadi ◽  
Maryam Lale Ataei ◽  
Esmaeil Ebrahimie ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Tumor Cells ◽  
Molecular Mechanisms ◽  
Tumor Development ◽  
Tumor Stroma ◽  
Cell Types ◽  
Great Promise ◽  
Tumor Tissues ◽  
Essential Components

Cancer has long been considered as a heterogeneous population of uncontrolled proliferation ofdifferent transformed cell types. The recent findings concerning tumorigeneses have highlightedthe fact that tumors can progress through tight relationships among tumor cells, cellular, andnon-cellular components which are present within tumor tissues. In recent years, studies haveshown that mesenchymal stem cells (MSCs) are essential components of non-tumor cells withinthe tumor tissues that can strongly affect tumor development. Several forms of MSCs have beenidentified within tumor stroma. Naïve (innate) mesenchymal stem cells (N-MSCs) derived fromdifferent sources are mostly recruited into the tumor stroma. N-MSCs exert dual and divergenteffects on tumor growth through different conditions and factors such as toll-like receptorpriming (TLR-priming), which is the primary underlying causes of opposite effects. Moreover,MSCs also have the contrary effects by various molecular mechanisms relying on direct cellto-cell connections and indirect communications through the autocrine, paracrine routes, andtumor microenvironment (TME).Overall, cell-based therapies will hold great promise to provide novel anticancer treatments.However, the application of intact MSCs in cancer treatment can theoretically cause adverseclinical outcomes. It is essential that to extensively analysis the effective factors and conditionsin which underlying mechanisms are adopted by MSCs when encounter with cancer.The aim is to review the cellular and molecular mechanisms underlying the dual effects ofMSCs followed by the importance of polarization of MSCs through priming of TLRs.<br />

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