Pellegrino M, Orsini P, Tognoni E
Istituto Nazionale di Ottica, Consiglio Nazionale delle Ricerche (INO-CNR), Via Moruzzi 1, Pisa 56124, Italy.
Dipartimento di Ricerca Traslazionale e delle Nuove Tecnologie in Medicina e Chirurgia, Università di Pisa, via S. Zeno 31, Pisa 56127, Italy.
Micron. 2024 Jun;181:103635. doi: 10.1016/j.micron.2024.103635. Epub 2024 Mar 28.
Human red blood cells show submicron membrane fluctuations (CMFs) that have been mainly studied with optical microscopies. Although the functional role of this phenomenon is still uncertain, the amplitude of membrane fluctuations is considered as an indicator of mechanical resilience to the stress encountered in the capillary beds. We investigate here the membrane fluctuations in red blood cells using the scanning ion conductance microscopy (SICM), a scanning probe technique that avoids the probe-sample contact. The ion current noise was recorded at a fixed distance from the cell and converted in terms of membrane fluctuation amplitude using as a converting factor the slope of the current-distance curve. We found that CMF amplitude was irreversibly reduced by membrane cross-link. Both whole cell and local increase of membrane tension induced a reduction of CMF amplitude. As for the biochemical regulation of membrane dynamics, we observed that the activation of the noradrenergic transduction pathway, via β-receptors, increased the CMF amplitude. We conclude that the CMFs recorded by SICM and those optically recorded on red blood cells share the main features. In addition SICM provides high spatial and temporal resolution as well as the possibility to apply through the glass pipette acting as probe chemical or physical stimuli to the membrane area where the CMFs are recorded.
人类红细胞表现出亚微米级的膜波动(CMFs),这主要是通过光学显微镜进行研究的。尽管这种现象的功能作用仍不确定,但膜波动的幅度被视为对毛细血管床中所遇应力的机械弹性指标。我们在此使用扫描离子电导显微镜(SICM)研究红细胞中的膜波动,SICM是一种避免探针与样品接触的扫描探针技术。在距细胞固定距离处记录离子电流噪声,并使用电流 - 距离曲线的斜率作为转换因子将其转换为膜波动幅度。我们发现膜交联会使CMF幅度不可逆地降低。全细胞膜张力和局部膜张力增加均会导致CMF幅度降低。至于膜动力学的生化调节,我们观察到通过β受体激活去甲肾上腺素能转导途径会增加CMF幅度。我们得出结论,SICM记录的CMFs与光学记录在红细胞上的CMFs具有主要特征。此外,SICM提供了高空间和时间分辨率,以及通过用作探针的玻璃微吸管对记录CMFs的膜区域施加化学或物理刺激的可能性。