Droplet Microfluidic Technology for the Early and Label-Free Isolation of Highly-Glycolytic, Activated T-Cells
A label-free, fixation-free and passive sorting method is presented to isolate activated T-cells shortly after activation and prior to the display of activation surface markers. It uses a recently developed sorting platform dubbed “Sorting by Interfacial Tension” (SIFT) that sorts droplets based on...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-09-01
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| Series: | Micromachines |
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| Online Access: | https://www.mdpi.com/2072-666X/13/9/1442 |
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| author | Claudia Zielke Adriana J. Gutierrez Ramirez Kelsey Voss Maya S. Ryan Azam Gholizadeh Jeffrey C. Rathmell Paul Abbyad |
| author_facet | Claudia Zielke Adriana J. Gutierrez Ramirez Kelsey Voss Maya S. Ryan Azam Gholizadeh Jeffrey C. Rathmell Paul Abbyad |
| author_sort | Claudia Zielke |
| collection | DOAJ |
| description | A label-free, fixation-free and passive sorting method is presented to isolate activated T-cells shortly after activation and prior to the display of activation surface markers. It uses a recently developed sorting platform dubbed “Sorting by Interfacial Tension” (SIFT) that sorts droplets based on pH. After polyclonal (anti-CD3/CD28 bead) activation and a brief incubation on chip, droplets containing activated T-cells display a lower pH than those containing naive cells due to increased glycolysis. Under specific surfactant conditions, a change in pH can lead to a concurrent increase in droplet interfacial tension. The isolation of activated T-cells on chip is hence achieved as flattened droplets are displaced as they encounter a micro-fabricated trench oriented diagonally with respect to the direction of flow. This technique leads to an enrichment of activated primary CD4+ T-cells to over 95% from an initial mixed population of naive cells and cells activated for as little as 15 min. Moreover, since the pH change is correlated to successful activation, the technique allows the isolation of T-cells with the earliest activation and highest glycolysis, an important feature for the testing of T-cell activation modulators and to determine regulators and predictors of differentiation outcomes. |
| first_indexed | 2024-03-09T23:07:32Z |
| format | Article |
| id | doaj.art-0ed9c24495f2457e8a39eb691ffdcbb7 |
| institution | Directory Open Access Journal |
| issn | 2072-666X |
| language | English |
| last_indexed | 2024-03-09T23:07:32Z |
| publishDate | 2022-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Micromachines |
| spelling | doaj.art-0ed9c24495f2457e8a39eb691ffdcbb72023-11-23T17:49:38ZengMDPI AGMicromachines2072-666X2022-09-01139144210.3390/mi13091442Droplet Microfluidic Technology for the Early and Label-Free Isolation of Highly-Glycolytic, Activated T-CellsClaudia Zielke0Adriana J. Gutierrez Ramirez1Kelsey Voss2Maya S. Ryan3Azam Gholizadeh4Jeffrey C. Rathmell5Paul Abbyad6Department of Chemistry and Biochemistry, Santa Clara University, Santa Clara, CA 95053, USADepartment of Chemistry and Biochemistry, Santa Clara University, Santa Clara, CA 95053, USADepartment of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, TN 37232, USADepartment of Chemistry and Biochemistry, Santa Clara University, Santa Clara, CA 95053, USADepartment of Chemistry and Biochemistry, Santa Clara University, Santa Clara, CA 95053, USADepartment of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, TN 37232, USADepartment of Chemistry and Biochemistry, Santa Clara University, Santa Clara, CA 95053, USAA label-free, fixation-free and passive sorting method is presented to isolate activated T-cells shortly after activation and prior to the display of activation surface markers. It uses a recently developed sorting platform dubbed “Sorting by Interfacial Tension” (SIFT) that sorts droplets based on pH. After polyclonal (anti-CD3/CD28 bead) activation and a brief incubation on chip, droplets containing activated T-cells display a lower pH than those containing naive cells due to increased glycolysis. Under specific surfactant conditions, a change in pH can lead to a concurrent increase in droplet interfacial tension. The isolation of activated T-cells on chip is hence achieved as flattened droplets are displaced as they encounter a micro-fabricated trench oriented diagonally with respect to the direction of flow. This technique leads to an enrichment of activated primary CD4+ T-cells to over 95% from an initial mixed population of naive cells and cells activated for as little as 15 min. Moreover, since the pH change is correlated to successful activation, the technique allows the isolation of T-cells with the earliest activation and highest glycolysis, an important feature for the testing of T-cell activation modulators and to determine regulators and predictors of differentiation outcomes.https://www.mdpi.com/2072-666X/13/9/1442microfluidicsdroplet microfluidicscytometrysortingpassive sortingT-cells |
| spellingShingle | Claudia Zielke Adriana J. Gutierrez Ramirez Kelsey Voss Maya S. Ryan Azam Gholizadeh Jeffrey C. Rathmell Paul Abbyad Droplet Microfluidic Technology for the Early and Label-Free Isolation of Highly-Glycolytic, Activated T-Cells Micromachines microfluidics droplet microfluidics cytometry sorting passive sorting T-cells |
| title | Droplet Microfluidic Technology for the Early and Label-Free Isolation of Highly-Glycolytic, Activated T-Cells |
| title_full | Droplet Microfluidic Technology for the Early and Label-Free Isolation of Highly-Glycolytic, Activated T-Cells |
| title_fullStr | Droplet Microfluidic Technology for the Early and Label-Free Isolation of Highly-Glycolytic, Activated T-Cells |
| title_full_unstemmed | Droplet Microfluidic Technology for the Early and Label-Free Isolation of Highly-Glycolytic, Activated T-Cells |
| title_short | Droplet Microfluidic Technology for the Early and Label-Free Isolation of Highly-Glycolytic, Activated T-Cells |
| title_sort | droplet microfluidic technology for the early and label free isolation of highly glycolytic activated t cells |
| topic | microfluidics droplet microfluidics cytometry sorting passive sorting T-cells |
| url | https://www.mdpi.com/2072-666X/13/9/1442 |
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