The label-free separation and culture of tumor cells in a microfluidic biochip

The Analyst ◽  
2020 ◽  
Vol 145 (5) ◽  
pp. 1706-1715 ◽  
Author(s):  
Jian Zhou ◽  
Chunlong Tu ◽  
Yitao Liang ◽  
Bobo Huang ◽  
Yifeng Fang ◽  
...  
Keyword(s):  

An integrated microfluidic biochip was designed for the streamlined separation, capture, in situ culture and/or immunofluorescence characterization of tumor cells.

2016 ◽  
Vol 8 (35) ◽  
pp. 22875-22883 ◽  
Author(s):  
Wei-Jiao Liu ◽  
Di Zhang ◽  
Li-Li Li ◽  
Zeng-Ying Qiao ◽  
Ju-Chen Zhang ◽  
...  

2020 ◽  
Vol 9 (4) ◽  
pp. 1206 ◽  
Author(s):  
Arvind Iyer ◽  
Krishan Gupta ◽  
Shreya Sharma ◽  
Kishore Hari ◽  
Yi Fang Lee ◽  
...  

We collated publicly available single-cell expression profiles of circulating tumor cells (CTCs) and showed that CTCs across cancers lie on a near-perfect continuum of epithelial to mesenchymal (EMT) transition. Integrative analysis of CTC transcriptomes also highlighted the inverse gene expression pattern between PD-L1 and MHC, which is implicated in cancer immunotherapy. We used the CTCs expression profiles in tandem with publicly available peripheral blood mononuclear cell (PBMC) transcriptomes to train a classifier that accurately recognizes CTCs of diverse phenotype. Further, we used this classifier to validate circulating breast tumor cells captured using a newly developed microfluidic system for label-free enrichment of CTCs.


2019 ◽  
Vol 116 (48) ◽  
pp. 24012-24018 ◽  
Author(s):  
Sixian You ◽  
Ronit Barkalifa ◽  
Eric J. Chaney ◽  
Haohua Tu ◽  
Jaena Park ◽  
...  

Despite extensive interest, extracellular vesicle (EV) research remains technically challenging. One of the unexplored gaps in EV research has been the inability to characterize the spatially and functionally heterogeneous populations of EVs based on their metabolic profile. In this paper, we utilize the intrinsic optical metabolic and structural contrast of EVs and demonstrate in vivo/in situ characterization of EVs in a variety of unprocessed (pre)clinical samples. With a pixel-level segmentation mask provided by the deep neural network, individual EVs can be analyzed in terms of their optical signature in the context of their spatial distribution. Quantitative analysis of living tumor-bearing animals and fresh excised human breast tissue revealed abundance of NAD(P)H-rich EVs within the tumor, near the tumor boundary, and around vessel structures. Furthermore, the percentage of NAD(P)H-rich EVs is highly correlated with human breast cancer diagnosis, which emphasizes the important role of metabolic imaging for EV characterization as well as its potential for clinical applications. In addition to the characterization of EV properties, we also demonstrate label-free monitoring of EV dynamics (uptake, release, and movement) in live cells and animals. The in situ metabolic profiling capacity of the proposed method together with the finding of increasing NAD(P)H-rich EV subpopulations in breast cancer have the potential for empowering applications in basic science and enhancing our understanding of the active metabolic roles that EVs play in cancer progression.


2021 ◽  
Vol 22 (10) ◽  
pp. 5068
Author(s):  
Igor Buzalewicz ◽  
Agnieszka Ulatowska-Jarża ◽  
Aleksandra Kaczorowska ◽  
Marlena Gąsior-Głogowska ◽  
Halina Podbielska ◽  
...  

Quantifying changes in bacteria cells in the presence of antibacterial treatment is one of the main challenges facing contemporary medicine; it is a challenge that is relevant for tackling issues pertaining to bacterial biofilm formation that substantially decreases susceptibility to biocidal agents. Three-dimensional label-free imaging and quantitative analysis of bacteria–photosensitizer interactions, crucial for antimicrobial photodynamic therapy, is still limited due to the use of conventional imaging techniques. We present a new method for investigating the alterations in living cells and quantitatively analyzing the process of bacteria photodynamic inactivation. Digital holographic tomography (DHT) was used for in situ examination of the response of Escherichia coli and Staphylococcus aureus to the accumulation of the photosensitizers immobilized in the copolymer revealed by the changes in the 3D refractive index distributions of single cells. Obtained results were confirmed by confocal microscopy and statistical analysis. We demonstrated that DHT enables real-time characterization of the subcellular structures, the biophysical processes, and the induced local changes of the intracellular density in a label-free manner and at sub-micrometer spatial resolution.


2009 ◽  
Vol 75A (6) ◽  
pp. 520-527 ◽  
Author(s):  
Joost F. Swennenhuis ◽  
Arjan G. J. Tibbe ◽  
Rianne Levink ◽  
Ronald C. J. Sipkema ◽  
Leon W. M. M. Terstappen

1992 ◽  
Vol 40 (2) ◽  
pp. 171-175 ◽  
Author(s):  
K Weber-Matthiesen ◽  
M Winkemann ◽  
A Müller-Hermelink ◽  
B Schlegelberger ◽  
W Grote

In immunocytochemical studies, the phenotypic evaluation of tumor cells is often complicated by accompanying normal cells, representing the original tissue or infiltrating leukocytes. This holds particularly true for tissues with a great morphological and immunophenotypical variability, such as bone marrow. A method that identifies mitotic tumor cells by chromosomal aberrations and permits the subsequent immunophenotypical analysis was a first progress, demonstrated by Teerenhovi et al. However, the results are usually hampered by the low number of analyzable mitoses. We demonstrate here a method that simultaneously combines immunophenotyping and in situ hybridization with centromere-specific probes. Using our method, numerically aberrant tumor cells can be identified by interphase cytogenetics and subsequently analyzed immunophenotypically. Since all interphase cells can be analyzed, we are not limited by the number and banding quality of analyzable mitoses.


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