Production of AFM wedged cantilevers for stress-relaxation experiments: Uniaxial loading of soft, spherical cells

IF 4.2 3区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

Methods Pub Date : 2025-02-17 DOI:10.1016/j.ymeth.2025.02.004

Riccardo Campanile , Jonne Helenius , Cristina Scielzo , Lydia Scarfò , Domenico Salerno , Mario Bossi , Marta Falappi , Alessia Saponara , Daniel J. Müller , Francesco Mantegazza , Valeria Cassina

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引用次数: 0

Abstract

The fabrication of wedge-shaped cantilevers for Atomic Force Microscopy (AFM) remains a critical yet challenging task, particularly when precision and efficiency are required. In this study, we present a streamlined protocol for producing these wedges using NOA63 UV-curing polymer, which simplifies the process and eliminates the need for dedicated equipment. Our method reduces preparation time while maintaining the mechanical properties of the cantilevers, in line with the manufacturer's specifications. We demonstrate the effectiveness of our wedged cantilevers in stress-relaxation experiments performed by means of AFM and confocal microscopy on primary Chronic Lymphocytic Leukemia cells and the MEC1 cell line. These experiments highlight the effectiveness of using modified cantilevers to consistently apply precise uniaxial loading to soft, spherical cells. This technique offers a marked improvement in fabrication speed and operational ease compared to traditional methods, without compromising the accuracy or performance of the measurements. This protocol is not only time-saving, but also adaptable for use in a wide range of biological applications, making it a valuable tool for AFM-based research in cellular mechanics.

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来源期刊

Methods 生物-生化研究方法

CiteScore

9.80

自引率

2.10%

发文量

222

审稿时长

11.3 weeks

期刊介绍： Methods focuses on rapidly developing techniques in the experimental biological and medical sciences. Each topical issue, organized by a guest editor who is an expert in the area covered, consists solely of invited quality articles by specialist authors, many of them reviews. Issues are devoted to specific technical approaches with emphasis on clear detailed descriptions of protocols that allow them to be reproduced easily. The background information provided enables researchers to understand the principles underlying the methods; other helpful sections include comparisons of alternative methods giving the advantages and disadvantages of particular methods, guidance on avoiding potential pitfalls, and suggestions for troubleshooting.