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基于圖形化彈性基底的細(xì)胞牽引力測(cè)量研究
電子技術(shù)應(yīng)用 2023年3期
徐紅梅1,,張帆1,,杜曉晗1,樊文強(qiáng)1,,周贇2,,陳修寰2,,趙冠棋2,朱疆1
(1.北京信息科技大學(xué) 光電測(cè)試技術(shù)及儀器教育部重點(diǎn)實(shí)驗(yàn)室,, 北京 100192,; 2.首都醫(yī)科大學(xué)附屬北京安貞醫(yī)院, 北京 100029)
摘要: 細(xì)胞牽引力顯微鏡方法是測(cè)量細(xì)胞牽引力的主流工具之一,,該方法一般使用熒光微珠作為標(biāo)志點(diǎn),,通過(guò)測(cè)量細(xì)胞牽引力作用下熒光微珠的位移場(chǎng),反演得出細(xì)胞牽引力場(chǎng),。由于熒光微珠在基底分布的深度不可控,,但計(jì)算時(shí)候默認(rèn)所有微珠分布在基底表面,一定程度上降低了細(xì)胞牽引力場(chǎng)的計(jì)算精度,。針對(duì)熒光微珠深度不可控的問(wèn)題,,提出在彈性基底表面加工圖形陣列作為基底變形標(biāo)志點(diǎn),開(kāi)展細(xì)胞牽引力測(cè)量的方法,。在聚二甲基硅氧烷基底表面設(shè)計(jì)直徑3 μm,, 高度0.8 μm的凸臺(tái)陣列作為基底位移標(biāo)志點(diǎn)。使用有限元仿真軟件在圖形化基底表面不同位置施加外力,,仿真得到凸臺(tái)陣列的位移場(chǎng),,利用牽引力反演算法計(jì)算得到力場(chǎng),與仿真過(guò)程中輸入的外力場(chǎng)相比較,,結(jié)果一致,。使用乳鼠心肌細(xì)胞在設(shè)計(jì)加工的圖形化彈性基底上開(kāi)展了牽引力測(cè)量實(shí)驗(yàn),,并計(jì)算得出牽引力場(chǎng)。仿真和實(shí)驗(yàn)均表明圖形化彈性基底適用于細(xì)胞牽引力的測(cè)量,,為心肌相關(guān)疾病病理研究提供了新的研究手段,。
中圖分類(lèi)號(hào):Q2-33 文獻(xiàn)標(biāo)志碼:A DOI: 10.16157/j.issn.0258-7998.223556
中文引用格式: 徐紅梅,張帆,,杜曉晗,,等. 基于圖形化彈性基底的細(xì)胞牽引力測(cè)量研究[J]. 電子技術(shù)應(yīng)用,2023,,49(3):30-36.
英文引用格式: Xu Hongmei,,Zhang Fan,Du Xiaohan,,et al. Cellular traction force measurement based on patterned elastic substrate[J]. Application of Electronic Technique,,2023,49(3):30-36.
Cellular traction force measurement based on patterned elastic substrate
Xu Hongmei1,,Zhang Fan1,,Du Xiaohan1,F(xiàn)an Wenqiang1,,Zhou Xun2,,Chen Xiuhuan2,Zhao Guanqi2,, Zhu Jiang1
(1.Key Laboratory of the Ministry of Education for Optoelectronic Measurement Technology and Instrument,, Beijing Information Science &Technology University, Beijing 100192,, China,; 2.Beijing Anzhen Hospital, Capital Medical University,, Beijing 100029, China)
Abstract: Abstract: Cell traction force microscopy method is one of the mainstream tools for measuring cell traction force, which uses fluorescent microbeads as markers to measure the deformation of the substrate and calculate cell traction force. However, due to the random distribution of fluorescent microbeads in the substrate, the calculation accuracy of the cell traction force field maybe affected. To address the problem of uncontrollable depth of fluorescent microbeads, a method using patterned elastic substrate was proposed to measure cell traction forces, in which the micropattern arrays fabricated on the surface of substrate was used as deformation markers. A micro-pad array with diameter of 3 µm and height of 0.8 µm was designed and fabricated on the polydimethylsiloxane substrate surface as the displacement markers. In the finite element simulation, external forces mimicking cell traction forces was applied at different locations on the substrate surface and the displacements field of the micro-pad array was obtained. Using the displacements field obtained by simulation, the traction force field was calculated through the traction force inversion algorithm, and the results were consistent with the input external forces in the simulation. The patterned elastic substrates were validated by mapping neonatal rat ventricular myocytes contraction forces. Both simulation and experiment demonstrate that the capacity of patterned elastic substrate for generating accurate cell contraction force maps, providing new research method for pathological study of myocardial diseases.
Key words : cell traction force mapping,;patterned substrates,;finite element simulation;traction force inversion

0 引言

早期的細(xì)胞力學(xué)主要研究外力如何作用于細(xì)胞的問(wèn)題,。時(shí)至今日,,細(xì)胞力學(xué)已從研究單個(gè)細(xì)胞力學(xué)性質(zhì)發(fā)展到關(guān)注細(xì)胞與細(xì)胞、細(xì)胞與基底的相互作用,。了解細(xì)胞如何產(chǎn)生和感知力,,以及這些力如何轉(zhuǎn)化為生化信號(hào),對(duì)于解決關(guān)于正常和病理狀態(tài)下細(xì)胞行為的基本問(wèn)題至關(guān)重要,。如何精確地測(cè)量細(xì)胞對(duì)外界施加的力是生物力學(xué)研究中的關(guān)鍵問(wèn)題,。

目前,,細(xì)胞牽引力測(cè)量的兩大主流方法有微柱陣列法和細(xì)胞牽引力顯微鏡法(Cell Traction Force Microscopy,CTFM),。利用微柱陣列的方法來(lái)測(cè)量細(xì)胞牽引力,,優(yōu)勢(shì)在于微柱的力學(xué)模型簡(jiǎn)單,算法簡(jiǎn)單,。但是當(dāng)微柱的間距超過(guò)懸浮細(xì)胞的尺寸時(shí),,在微柱上接種細(xì)胞時(shí),細(xì)胞會(huì)落入微柱之間,,黏附在基底上,,無(wú)法進(jìn)行細(xì)胞牽引力的測(cè)量。CTFM方法的優(yōu)勢(shì)在于基底連續(xù),,制備方法簡(jiǎn)單,,基底一般是熒光微珠和凝膠混合制備;缺點(diǎn)在于基底位移場(chǎng)和細(xì)胞牽引力場(chǎng)高度耦合,,計(jì)算較為復(fù)雜,。CTFM方法使用的彈性基底是熒光微珠和凝膠的混合物,熒光微珠分布在凝膠的不同深度,,但在解算位移場(chǎng)時(shí)對(duì)微珠所處的深度沒(méi)有加以考慮,,導(dǎo)致位移場(chǎng)計(jì)算精度受到一定影響。本文通過(guò)Comsol軟件仿真在基底表面模擬細(xì)胞施加150 nN的外力,,得到距離基底表面不同深度的微珠位移量,,因微珠分布不均勻?qū)е掠?jì)算位移場(chǎng)的偏差在0.1μm左右。所以微珠分布在基底的不同深度會(huì)影響位移場(chǎng)的計(jì)算精度,。2016年,,Martin Bergert提出使用量子點(diǎn)(QDs)納米滴印方法,在彈性基底表面形成了六邊形規(guī)律排布的量子點(diǎn)陣列,,避免了位移標(biāo)志點(diǎn)分布隨機(jī),、深度不均勻的問(wèn)題。但量子點(diǎn)陣列制備工藝復(fù)雜,,對(duì)細(xì)胞有一定毒性,。

因此,針對(duì)細(xì)胞牽引力顯微鏡方法中熒光微珠深度不可控的問(wèn)題,,本文提出在彈性基底表面加工微圖形陣列代替熒光微珠作為標(biāo)志點(diǎn)的方法,,開(kāi)展細(xì)胞牽引力測(cè)量實(shí)驗(yàn)。本文首先介紹了牽引力場(chǎng)測(cè)量的原理,;其次,,對(duì)設(shè)計(jì)有微凸臺(tái)陣列的彈性基底進(jìn)行有限元仿真,獲得彈性基底在模擬細(xì)胞力作用下產(chǎn)生的位移場(chǎng),,通過(guò)牽引力反演算法將仿真得到的位移場(chǎng)反演,,計(jì)算得出力場(chǎng),,通過(guò)比較仿真過(guò)程中輸入的力場(chǎng)與牽引力反演算法計(jì)算得出的力場(chǎng)是否相等,驗(yàn)證基底設(shè)計(jì)和算法的有效性,;最后,,在圖形化彈性基底上接種原代乳鼠心肌細(xì)胞,并利用細(xì)胞顯微成像和圖像處理技術(shù)測(cè)量微凸臺(tái)陣列的位移場(chǎng),,利用牽引力反演算法計(jì)算得到心肌細(xì)胞收縮力的大小和分布,。




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作者信息:

徐紅梅1,張帆1,,杜曉晗1,,樊文強(qiáng)1,周贇2,,陳修寰2,,趙冠棋2,朱疆1

(1.北京信息科技大學(xué) 光電測(cè)試技術(shù)及儀器教育部重點(diǎn)實(shí)驗(yàn)室,, 北京 100192,;

2.首都醫(yī)科大學(xué)附屬北京安貞醫(yī)院, 北京 100029)


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