Label‐Free Metabolic Classification of Single Cells in Droplets Using the Phasor Approach to Fluorescence Lifetime Imaging Microscopy

Ma, Ning; Kamalakshakurup, Gopakumar; Aghaamoo, Mohammad; Lee, Abraham P.; Digman, Michelle A.

doi:10.1002/cyto.a.23673

Cited by 7 publications

(7 citation statements)

References 53 publications

(60 reference statements)

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“…Because the primers are fixed in the droplet, the amplification efficiency of the target (Chiu et al, 2015). (B) Label-free metabolic classification of single cells in droplets using the phasor approach to fluorescence lifetime imaging microscopy (Ma et al, 2019).…”

Section: Single Cell Genomics Analysismentioning

confidence: 99%

“…(A) Schematic diagram of a microfluid based optical sensing device for labeling free detection of circulating tumor cells ( Chiu et al, 2015 ). (B) Label‐free metabolic classification of single cells in droplets using the phasor approach to fluorescence lifetime imaging microscopy ( Ma et al, 2019 ).…”

Section: Single-cell Analysis Technique Based On Droplet Microfluidicsmentioning

confidence: 99%

“…Due to increased glycolysis, tumor cells had a higher free/bound NADH ratio than normal cells. Ma et al (2019) combined droplets with fluorescence imaging methods, encapsulated single cells in droplets, and measured NADH metabolism changes to distinguish different leukemia cell types and stages of the cell cycle. The microfluidic device has an expansion flow focusing geometry as shown in Figure 6B .…”

Section: Single-cell Analysis Technique Based On Droplet Microfluidicsmentioning

confidence: 99%

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Droplets microfluidics platform—A tool for single cell research

Cheng

et al. 2023

Front. Bioeng. Biotechnol.

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Cells are the most basic structural and functional units of living organisms. Studies of cell growth, differentiation, apoptosis, and cell-cell interactions can help scientists understand the mysteries of living systems. However, there is considerable heterogeneity among cells. Great differences between individuals can be found even within the same cell cluster. Cell heterogeneity can only be clearly expressed and distinguished at the level of single cells. The development of droplet microfluidics technology opens up a new chapter for single-cell analysis. Microfluidic chips can produce many nanoscale monodisperse droplets, which can be used as small isolated micro-laboratories for various high-throughput, precise single-cell analyses. Moreover, gel droplets with good biocompatibility can be used in single-cell cultures and coupled with biomolecules for various downstream analyses of cellular metabolites. The droplets are also maneuverable; through physical and chemical forces, droplets can be divided, fused, and sorted to realize single-cell screening and other related studies. This review describes the channel design, droplet generation, and control technology of droplet microfluidics and gives a detailed overview of the application of droplet microfluidics in single-cell culture, single-cell screening, single-cell detection, and other aspects. Moreover, we provide a recent review of the application of droplet microfluidics in tumor single-cell immunoassays, describe in detail the advantages of microfluidics in tumor research, and predict the development of droplet microfluidics at the single-cell level.

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Section: Single Cell Genomics Analysismentioning

confidence: 99%

Section: Single-cell Analysis Technique Based On Droplet Microfluidicsmentioning

confidence: 99%

Section: Single-cell Analysis Technique Based On Droplet Microfluidicsmentioning

confidence: 99%

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Droplets microfluidics platform—A tool for single cell research

Cheng

et al. 2023

Front. Bioeng. Biotechnol.

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“…Other examples of this approach include the quantification of metabolic intrinsic fluorescent co-factors, such as nicotinamide adenine dinucleotide (NADH) and nicotinamide adenine dinucleotide phosphate (NADPH). This label-free method allowed researchers to distinguish the differential metabolic states of K-562 and Jurkat cell lines [ 32 ].…”

Section: Fluorescence-based Biosensorsmentioning

confidence: 99%

Optical Detection of Cancer Cells Using Lab-on-a-Chip

et al. 2023

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The global need for accurate and efficient cancer cell detection in biomedicine and clinical diagnosis has driven extensive research and technological development in the field. Precision, high-throughput, non-invasive separation, detection, and classification of individual cells are critical requirements for successful technology. Lab-on-a-chip devices offer enormous potential for solving biological and medical problems and have become a priority research area for microanalysis and manipulating cells. This paper reviews recent developments in the detection of cancer cells using the microfluidics-based lab-on-a-chip method, focusing on describing and explaining techniques that use optical phenomena and a plethora of probes for sensing, amplification, and immobilization. The paper describes how optics are applied in each experimental method, highlighting their advantages and disadvantages. The discussion includes a summary of current challenges and prospects for cancer diagnosis.

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“…A robust assay using droplet microfluidic technology, developed to study cell heterogeneity within and among cell lines, has been extended to characterize metabolic differences between proliferating and quiescent cells-a critical step toward labelfree single cancer cell dormancy research. This noninvasive droplet-based and label-free method, using an expansion flow-focusing geometry to distinguish individual cells based on their metabolic states, could therefore be used as an upstream phenotypic platform to correlate with genomic statistics [212]. The non-proliferative cellular quiescence state is required for cell survival under stress and during development.…”

Section: Microfluidics For Quiescence Researchmentioning

confidence: 99%

Application of Microfluidic Systems for Breast Cancer Research

et al. 2022

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Cancer is a disease in which cells in the body grow out of control; breast cancer is the most common cancer in women in the United States. Due to early screening and advancements in therapeutic interventions, deaths from breast cancer have declined over time, although breast cancer remains the second leading cause of cancer death among women. Most deaths are due to metastasis, as cancer cells from the primary tumor in the breast form secondary tumors in remote sites in distant organs. Over many years, the basic biological mechanisms of breast cancer initiation and progression, as well as the subsequent metastatic cascade, have been studied using cell cultures and animal models. These models, although extremely useful for delineating cellular mechanisms, are poor predictors of physiological responses, primarily due to lack of proper microenvironments. In the last decade, microfluidics has emerged as a technology that could lead to a paradigm shift in breast cancer research. With the introduction of the organ-on-a-chip concept, microfluidic-based systems have been developed to reconstitute the dominant functions of several organs. These systems enable the construction of 3D cellular co-cultures mimicking in vivo tissue-level microenvironments, including that of breast cancer. Several reviews have been presented focusing on breast cancer formation, growth and metastasis, including invasion, intravasation, and extravasation. In this review, realizing that breast cancer can recur decades following post-treatment disease-free survival, we expand the discussion to account for microfluidic applications in the important areas of breast cancer detection, dormancy, and therapeutic development. It appears that, in the future, the role of microfluidics will only increase in the effort to eradicate breast cancer.

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Label‐Free Metabolic Classification of Single Cells in Droplets Using the Phasor Approach to Fluorescence Lifetime Imaging Microscopy

Cited by 7 publications

References 53 publications

Droplets microfluidics platform—A tool for single cell research

Droplets microfluidics platform—A tool for single cell research

Optical Detection of Cancer Cells Using Lab-on-a-Chip

Application of Microfluidic Systems for Breast Cancer Research

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