Nanomechanical and Morphological AFM Mapping of Normal Tissues and Tumors on Live Brain Slices Using Specially Designed Embedding Matrix and Laser-Shaped Cantilevers.
Nanomechanical and Morphological AFM Mapping of Normal Tissues and Tumors on Live Brain Slices Using Specially Designed Embedding Matrix and Laser-Shaped Cantilevers.
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DOI：
10.3390/biomedicines10071742
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发表时间：
2022-07-19
期刊：
影响因子：
4.7
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中科院分区：
文献类型：
--
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细胞和组织纳米力学已被引入生物医学研究，不仅补充了传统的免疫分型和分子分析，而且还为临床诊断和生物工程带来了意想不到的新见解。然而，尽管原子力显微镜（AFM）在培养的单个细胞的研究中取得了进展，但其用于绘制活组织的应用存在许多技术限制。在这里，我们阐述了一种新的技术来研究正常脑组织和肿瘤的活切片相结合的形态学和纳米力学AFM映射在高通量扫描模式，在对比通常使用的力谱模式的基础上单点探针应用。这成为可能，由于结合使用适当的嵌入矩阵的振动和最初修改的AFM探针。通过调节琼脂和胶原蛋白I的量来仔细开发包埋基质组合物，以达到最佳的粘弹性，从而获得满足AFM要求的高质量活切片。AFM针尖圆形照射他们与聚焦纳秒激光脉冲，而所得的尖端形态进行了验证，通过扫描电子显微镜。活体切片制备和AFM研究仅需55分钟，并可与活细胞示踪分析或免疫染色相结合，从而使其有望用于生物医学研究和临床诊断。
Cell and tissue nanomechanics has been intriguingly introduced into biomedical research, not only complementing traditional immunophenotyping and molecular analysis, but also bringing unexpected new insights for clinical diagnostics and bioengineering. However, despite the progress in the study of individual cells in culture by atomic force microscopy (AFM), its application for mapping live tissues has a number of technical limitations. Here, we elaborate a new technique to study live slices of normal brain tissue and tumors by combining morphological and nanomechanical AFM mapping in high throughput scanning mode, in contrast to the typically utilized force spectroscopy mode based on single-point probe application. This became possible due to the combined use of an appropriate embedding matrix for vibratomy and originally modified AFM probes. The embedding matrix composition was carefully developed by regulating the amounts of agar and collagen I to reach optimal viscoelastic properties for obtaining high-quality live slices that meet AFM requirements. AFM tips were rounded by irradiating them with focused nanosecond laser pulses, while the resulting tip morphology was verified by scanning electron microscopy. Live slices preparation and AFM investigation take only 55 min and could be combined with a vital cell tracer analysis or immunostaining, thus making it promising for biomedical research and clinical diagnostics.
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影响因子：
16.6
作者：
Moeendarbary E;Weber IP;Sheridan GK;Koser DE;Soleman S;Haenzi B;Bradbury EJ;Fawcett J;Franze K
通讯作者：
Franze K
影响因子：
1.4
作者：
Grunow, Bianka;Kirchhoff, Tina;Harzsch, Steffen
通讯作者：
Harzsch, Steffen
影响因子：
1.2
作者：
Canovic, Elizabeth Peruski;Qing, Bo;Van Vliet, Krystyn J.
通讯作者：
Van Vliet, Krystyn J.
影响因子：
3
作者：
HAYARAN, A;BIJLANI, V
通讯作者：
BIJLANI, V
影响因子：
3.8
作者：
Migliorini, Elisa;Ban, Jelena;Lazzarino, Marco
通讯作者：
Lazzarino, Marco