Properties of cells through life and death - an acoustic microscopy investigation
Properties of cells through life and death - an acoustic microscopy investigation
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DOI：
10.1080/15384101.2015.1069925
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发表时间：
2015-09-17
期刊：
影响因子：
4.3
通讯作者：
Kolios, Michael C.
中科院分区：
文献类型：
--
作者：
Pasternak, Maurice M.;Strohm, Eric M.;Kolios, Michael C.
目前评估细胞状态的方法主要集中在分子生物标志物的荧光鉴定上。这些方法的侵入性——需要固定、化学染料、基因改变或这些方法的组合——阻碍了对样品的后续分析。鉴于这一限制，研究已经考虑使用物理标记来区分细胞阶段。声学显微镜是一种超高频（>100MHz）超声技术，可用于实时计算生物细胞的力学和物理特性，从而在没有侵入性生物标志物评估的情况下评估活细胞中的细胞阶段。使用声学显微镜，除了早期和晚期程序性细胞死亡外，还检查了增殖细胞周期G1、G2和中期的MCF-7人乳腺腺癌细胞。计算的物理性质包括胞体高度、声速、声阻抗、胞体密度、绝热体模量和超声衰减。总共测量了290个细胞，每个细胞期58个，使用荧光和相差显微镜进行评估。从G1向中期积极进展的细胞的衰减降低了28%，而G1诱导的细胞凋亡的衰减则显著增加了81%。此外，根据超声衰减，晚期凋亡细胞可分为2个不同的组，提示晚期凋亡可能存在目前尚未确定的亚阶段。一种不使用化学染料、固定或基因操作的细胞分期鉴定方法已被实施。
Current methods to evaluate the status of a cell are largely focused on fluorescent identification of molecular biomarkers. The invasive nature of these methods - requiring either fixation, chemical dyes, genetic alteration, or a combination of these - prevents subsequent analysis of samples. In light of this limitation, studies have considered the use of physical markers to differentiate cell stages. Acoustic microscopy is an ultrahigh frequency (>100MHz) ultrasound technology that can be used to calculate the mechanical and physical properties of biological cells in real-time, thereby evaluating cell stage in live cells without invasive biomarker evaluation. Using acoustic microscopy, MCF-7 human breast adenocarcinoma cells within the G1, G2, and metaphase phases of the proliferative cell cycle, in addition to early and late programmed cell death, were examined. Physical properties calculated include the cell height, sound speed, acoustic impedance, cell density, adiabatic bulk modulus, and the ultrasonic attenuation. A total of 290 cells were measured, 58 from each cell phase, assessed using fluorescent and phase contrast microscopy. Cells actively progressing from G1 to metaphase were marked by a 28% decrease in attenuation, in contrast to the induction of apoptosis from G1, which was marked by a significant 81% increase in attenuation. Furthermore late apoptotic cells separated into 2 distinct groups based on ultrasound attenuation, suggesting that presently-unidentified sub-stages may exist within late apoptosis. A methodology has been implemented for the identification of cell stages without the use of chemical dyes, fixation, or genetic manipulation.