Laser-fabricated cell patterning stencil for single cell analysis

Jacob J Messner,Deirdre R Meldrum,Honor L Glenn

doi:10.1186/s12896-017-0408-8

Jacob J Messner, Deirdre R Meldrum + Show 1 more

Open Access

https://doi.org/10.1186/s12896-017-0408-8

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Journal: BMC Biotechnology	Publication Date: Dec 1, 2017
Citations: 10	License type: open-access

Affiliation: Arizona State University

Abstract

Precise spatial positioning and isolation of mammalian cells is a critical component of many single cell experimental methods and biological engineering applications. Although a variety of cell patterning methods have been demonstrated, many of these methods subject cells to high stress environments, discriminate against certain phenotypes, or are a challenge to implement. Here, we demonstrate a rapid, simple, indiscriminate, and minimally perturbing cell patterning method using a laser fabricated polymer stencil. The stencil fabrication process requires no stencil-substrate alignment, and is readily adaptable to various substrate geometries and experiments.

Highlights

The ability to manipulate and selectively localize cells into patterns or distinct microenvironments is critical for single cell analysis [1,2,3,4], tissue engineering [5, 6], cell signaling studies [7,8,9], drug screening [10,11,12], and cell migration assays [13, 14]
We compared viability of cells seeded through the stencil to cells seeded directly onto an uncovered microwell array
Stencils are readily adaptable to specific substrates or experimentally desirable geometries and multiple cell types

Summary

Introduction

The ability to manipulate and selectively localize cells into patterns or distinct microenvironments is critical for single cell analysis [1,2,3,4], tissue engineering [5, 6], cell signaling studies [7,8,9], drug screening [10,11,12], and cell migration assays [13, 14]. Many active cell patterning and isolation methods utilize microfluidic systems, in which cells are manipulated and transported using fluidic forces. Inkjet-based cell ‘printing’ and deposition methods have proven effective at sorting and patterning cells at the bulk and single cell level, but are typically low throughput and raise concerns about cell stress responses [19,20,21,22]. While trap-based approaches are very high throughput, they may discriminate against particular cell morphologies or sizes with relevance for human disease [30]. Microfluidic trap environments impose difficulties in delivering single cells to isolated microenvironments for further experimentation

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Conclusion