Modifications to the Transwell Migration/Invasion Assay Method That Eases Assay Performance and Improves the Accuracy.

IF 1.6 4区医学 Q4 BIOCHEMICAL RESEARCH METHODS

Assay and drug development technologies Pub Date : 2022-02-01 DOI:10.1089/adt.2021.140

Heidi Marie Stoellinger, Arshak R Alexanian

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引用次数: 4

Abstract

Migration is a key property of live cells and critical for normal development, immune response, and disease processes such as cancer metastasis and inflammation. Methods to examine cell migration are especially useful and important for a wide range of biomedical research such as cancer biology, immunology, vascular biology, cell biology, and developmental biology. In vitro assays are excellent approaches to extrapolate to in vivo situations and study live cells behavior. The aim of this article is to discuss the existing methods for transwell migration/invasion studies, the problems associated with this assay, and proposed modifications to this methodological approach that makes it simple to perform and improve the assay accuracy. Results of our studies demonstrated that the count of cells that had grown on top of the membrane is important to accurately evaluate the percentage of migrated/invaded cells. The results also showed that the transparent transwell insert with 4',6-diamidino-2-phenylindole (DAPI) stained cells is the best approach to ease the analysis of cell numbers on top of the membranes. In addition, the overlay of bright light (representing membrane pores) and DAPI images can further improve the accuracy of cell count. All these modifications in combination simplify the assay performance and improve the accuracy of the transwell migration assay method.

Abstract Image

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对Transwell迁移/入侵分析方法的改进，简化了分析性能并提高了准确性。

迁移是活细胞的一个关键特性，对正常发育、免疫反应和癌症转移和炎症等疾病过程至关重要。检测细胞迁移的方法对于广泛的生物医学研究尤其有用和重要，如癌症生物学、免疫学、血管生物学、细胞生物学和发育生物学。体外分析是推断体内情况和研究活细胞行为的极好方法。本文的目的是讨论现有的跨井迁移/入侵研究方法，与该分析相关的问题，并提出对该方法方法的修改，使其易于执行并提高分析准确性。我们的研究结果表明，生长在膜上的细胞计数对于准确评估迁移/侵袭细胞的百分比很重要。结果还表明，4'，6-二氨基-2-苯基吲哚(DAPI)染色细胞的透明transwell插入是简化膜上细胞数量分析的最佳方法。此外，亮光(代表膜孔)与DAPI图像叠加可以进一步提高细胞计数的准确性。所有这些改进结合起来简化了分析性能，提高了跨井迁移分析方法的准确性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Assay and drug development technologies 医学-生化研究方法

CiteScore

3.60

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： ASSAY and Drug Development Technologies provides access to novel techniques and robust tools that enable critical advances in early-stage screening. This research published in the Journal leads to important therapeutics and platforms for drug discovery and development. This reputable peer-reviewed journal features original papers application-oriented technology reviews, topical issues on novel and burgeoning areas of research, and reports in methodology and technology application. ASSAY and Drug Development Technologies coverage includes: -Assay design, target development, and high-throughput technologies- Hit to Lead optimization and medicinal chemistry through preclinical candidate selection- Lab automation, sample management, bioinformatics, data mining, virtual screening, and data analysis- Approaches to assays configured for gene families, inherited, and infectious diseases- Assays and strategies for adapting model organisms to drug discovery- The use of stem cells as models of disease- Translation of phenotypic outputs to target identification- Exploration and mechanistic studies of the technical basis for assay and screening artifacts