Literature DB >> 30799837

A Pipette-Tip Based Method for Seeding Cells to Droplet Microfluidic Platforms.

Nidhi Sinha1, Nikita Subedi1, Florian Wimmers2, Melf Soennichsen1, Jurjen Tel3.   

Abstract

Amongst various microfluidic platform designs frequently used for cellular analysis, droplet-microfluidics provides a robust tool for isolating and analyzing cells at the single-cell level by eliminating the influence of external factors on the cellular microenvironment. Encapsulation of cells in droplets is dictated by the Poisson distribution as a function of the number of cells present in each droplet and the average number of cells per volume of droplet. Primary cells, especially immune cells, or clinical specimens can be scarce and loss-less encapsulation of cells remains challenging. In this paper, we present a new methodology that uses pipette-tips to load cells to droplet-based microfluidic devices without the significant loss of cells. With various cell types , we demonstrate efficient cell encapsulation in droplets that closely corresponds to the encapsulation efficiency predicted by the Poisson distribution. Our method ensures loss-less loading of cells to microfluidic platforms and can be easily adapted for downstream single cell analysis, e.g., to decode cellular interactions between different cell types.

Year:  2019        PMID: 30799837     DOI: 10.3791/57848

Source DB:  PubMed          Journal:  J Vis Exp        ISSN: 1940-087X            Impact factor:   1.355


  5 in total

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Authors:  B M Tiemeijer; J Tel
Journal:  Front Bioeng Biotechnol       Date:  2022-06-17

2.  Single-cell intracellular epitope and transcript detection reveals signal transduction dynamics.

Authors:  Francesca Rivello; Erik van Buijtenen; Kinga Matuła; Jessie A G L van Buggenum; Paul Vink; Hans van Eenennaam; Klaas W Mulder; Wilhelm T S Huck
Journal:  Cell Rep Methods       Date:  2021-09-15

3.  Enhancement of amino acid production and secretion by Lactococcus lactis using a droplet-based biosensing and selection system.

Authors:  Jhonatan A Hernandez-Valdes; Myrthe Aan de Stegge; Jos Hermans; Johan Teunis; Rinke J van Tatenhove-Pel; Bas Teusink; Herwig Bachmann; Oscar P Kuipers
Journal:  Metab Eng Commun       Date:  2020-06-04

4.  Hydrop enables droplet-based single-cell ATAC-seq and single-cell RNA-seq using dissolvable hydrogel beads.

Authors:  Suresh Poovathingal; Stein Aerts; Florian V De Rop; Joy N Ismail; Carmen Bravo González-Blas; Gert J Hulselmans; Christopher Campbell Flerin; Jasper Janssens; Koen Theunis; Valerie M Christiaens; Jasper Wouters; Gabriele Marcassa; Joris de Wit
Journal:  Elife       Date:  2022-02-23       Impact factor: 8.713

5.  Facile and scalable tubing-free sample loading for droplet microfluidics.

Authors:  Fangchi Shao; Kuangwen Hsieh; Pengfei Zhang; Aniruddha M Kaushik; Tza-Huei Wang
Journal:  Sci Rep       Date:  2022-08-03       Impact factor: 4.996

  5 in total

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