標(biāo)準(zhǔn)培養(yǎng)限制有幾種生理和病理過程，其中細(xì)胞經(jīng)歷體積變化。然而，沒有可靠的方法可用于實(shí)時(shí)準(zhǔn)確測量貼壁細(xì)胞的體積。 細(xì)胞體積測定的優(yōu)勢細(xì)胞在光學(xué)透明的腔室中培養(yǎng)，該腔室能夠間準(zhǔn)確確定它們的體積，并并行跟蹤導(dǎo)致體積變化的生化過程，例如離子泵的激活。 例子從間期到 Raji 細(xì)胞有絲分裂的體積跟蹤 [2]。
（A） 將細(xì)胞置于由柱設(shè)定的校準(zhǔn)高度的聚（二甲基硅氧烷）室中，在補(bǔ)充有FITC葡聚糖的培養(yǎng)基中。下圖：細(xì)胞在熒光圖像上排除熒光（比例尺20 mm）。 （B） 對應(yīng)于（A）中虛線的熒光文件：熒光強(qiáng)度的大值和小值分別對應(yīng)于腔室大高度（背景）和零高度（柱）。右圖：這些值用于校準(zhǔn)信號(hào)并計(jì)算細(xì)胞的光學(xué)厚度。 （C） 后，通過對細(xì)胞面積上的總熒光強(qiáng)度進(jìn)行積分來獲得細(xì)胞體積。 (A) HeLa 細(xì)胞的體積軌跡。開始和結(jié)束時(shí)的兩個(gè)體積超調(diào)對應(yīng)于有絲分裂中的瞬時(shí)體積增加，一個(gè)對應(yīng)于母細(xì)胞，二個(gè)對應(yīng)于子細(xì)胞。 (B) (A) 中使用 FXm 的細(xì)胞的原始熒光圖像。比例尺 50 毫米。
參考[1] Zlotek-Zlotkiewicz, E. 等人。(2015)。細(xì)胞生物學(xué)雜志，211 (4)，765–774。 [2] Cadart, C 等人。(2017)。細(xì)胞生物學(xué)方法，139,103-120。
提供實(shí)現(xiàn)工具細(xì)胞限制器-cell confiner（Cell confinement） 我們的cell Confiner是一種多功能設(shè)備，可通過對細(xì)胞應(yīng)用定義明確的約束條件來研究細(xì)胞力學(xué)。限制方法基于將細(xì)胞固定在兩個(gè)平行表面之間，從而實(shí)現(xiàn)均勻且定義明確的物理參數(shù)，例如細(xì)胞幾何形狀和環(huán)境彈性。此外，可以使用高分辨率顯微鏡對受限的細(xì)胞進(jìn)行成像，因?yàn)樵撛O(shè)備是光學(xué)透明的，并將細(xì)胞保持在焦平面上。 細(xì)胞被均勻地限制/壓縮在兩個(gè)亞微米分辨率的兩個(gè)平行表面之間。兩個(gè)表面之間的空間由微型PDMS支柱控制。 微型支柱在載玻片上制造，載玻片連接到PDMS活塞（吸盤）上。 活塞由真空泵控制，因此限制區(qū)的高度也受到控制。不同的限制高度（例如1um – 300um），允許長期細(xì)胞培養(yǎng)和細(xì)胞增殖，同時(shí)保持對封閉的完美控制
與高分辨率光學(xué)顯微鏡系統(tǒng)兼容，可以處理足夠多的細(xì)胞以進(jìn)行完整的基因表達(dá)分析，可與生物功能化的微結(jié)構(gòu)化底物和/或不同的基質(zhì)（幾何形狀控制）結(jié)合使用
可以與凝膠結(jié)合（硬度控制），兼容任何細(xì)胞培養(yǎng)底物（培養(yǎng)皿至96孔板）。 產(chǎn)品特性： >定義細(xì)胞的厚度和形狀 用正確的限制滑片控制細(xì)胞的厚度 >同時(shí)進(jìn)行多個(gè)實(shí)驗(yàn) 能夠研究不同的細(xì)胞或同時(shí)應(yīng)用不同的限制條件 >適用于高分辨率顯微鏡 光學(xué)透明的材料和緊湊的設(shè)計(jì)可實(shí)現(xiàn)高分辨率顯微鏡 >控制限制速度 通過真空泵j確控制限制速度 >可逆限制：限制后取回您的細(xì)胞 由于細(xì)胞的非破壞性方法，可以進(jìn)行分子分析 >與您自己的實(shí)驗(yàn)兼容 該限制器是一種小型設(shè)備，直接放置在您的細(xì)胞培養(yǎng)液頂部 技術(shù)方案圖
三種產(chǎn)品形式典型應(yīng)用：>癌癥浸潤測定：遷移行為和遷移轉(zhuǎn)變的量化 >癌癥侵襲性測定：體細(xì)胞或癌細(xì)胞的收縮力定量 >內(nèi)吞作用測定：更好地觀察膜發(fā)生的事件 >胞吐法測定：更好地觀察在頂端膜發(fā)生的事件 >吞噬功能失調(diào)：機(jī)制的表征 >孔中的免疫系統(tǒng)：非粘附免疫細(xì)胞的二維遷移和相互作用 >免疫細(xì)胞相互作用：非貼壁免疫細(xì)胞的2D相互作用 >有絲分裂組裝測定：有絲分裂紡錘體疾病的定量 >定量細(xì)胞遷移測定：細(xì)胞遷移特性的快速，精細(xì)分析 >癌癥研究 轉(zhuǎn)移細(xì)胞的遷移 轉(zhuǎn)移中的細(xì)胞收縮 DNA DSB修復(fù)（機(jī)械誘導(dǎo)） 基因組不穩(wěn)定（細(xì)胞分裂） 分離共培養(yǎng) >免疫學(xué) 免疫細(xì)胞遷移 非粘附細(xì)胞的成像 >器官生理學(xué) 癌細(xì)胞遷移 具有硬度控制的細(xì)胞區(qū)分 傷口愈合 分離共培養(yǎng) 細(xì)胞壓縮反應(yīng) >罕見疾病 細(xì)胞核完整性 >老化 細(xì)胞核完整性 自噬相關(guān)疾病 >觀測化 非粘附細(xì)胞的成像 細(xì)胞器的平面成像 >基礎(chǔ)研究 細(xì)胞體積（細(xì)胞周期） 細(xì)胞機(jī)械力刺激反應(yīng) 二維心肌細(xì)胞成熟測定
二維肝小管化驗(yàn)
3D心肌細(xì)胞成熟測定
3D肝小管測定
附著球體測定
細(xì)胞收縮力測定
細(xì)胞遷移測定
細(xì)胞核擠壓測定
細(xì)胞j化
細(xì)胞體積測量
趨化性測定
共培養(yǎng)測定
胞吞試驗(yàn)
胞吐法
外泌體測定
片狀脂蛋白和絲狀體含量測定
活細(xì)胞成像
巨噬細(xì)胞j化測定
MT依賴性運(yùn)輸測定
神經(jīng)肌肉連接測定
井中的神經(jīng)元網(wǎng)絡(luò)
細(xì)胞器定位分析
初次纖毛測定
骨骼肌細(xì)胞測定
平滑肌細(xì)胞
傷口愈合測定
PUBLICATIONS - Confinement and Low Adhesion Induce Fast Amoeboid Migration of Slow Mesenchymal Cells
Y.-J. Liu, M. Piel, Cell, et al., 2015 160(4), 659-672 - Actin flows induce a universal coupling between cell speed and cell persistence
P. Maiuri, R. Voituriez, et al., Cell, 2015 161(2), 374–386 - Geometric friction directs cell migration
M. Le Berre, M. Piel, et al., Physical Review Letter 2013 111, 198101 - Mitotic rounding alters cell geometry to ensure efficient spindle assembly
O. M. Lancaster, B. Baum, et al., Developmental Cell, 2013 25(3), 270-283 - Fine Control of Nuclear Confinement Identifies a Threshold Deformation leading to Lamina Rupture and Induction of Specific Genes
M. Le Berre, J. Aubertin, M. Piel, Integrative Biology, 2012 4 (11), 1406-1414 - Exploring the Function of Cell Shape and Size during Mitosis
C. Cadart, H. K. Matthews, et al., Developmental Cell, 2014 29(2), 159-169 - Methods for Two-Dimensional Cell Confinement
M. Le Berre, M. Piel, et al., 2014, Micropatterning in Cell Biology Part C, Methods in cell biology, 121, 213-29
References [1] D. Huh, G. A. Hamilton, and D. E. Ingber, “From 3D cell culture to organs-on-chips,” Trends Cell Biol., vol. 21, no. 12, pp. 745–754, 2011. [2] M. Ravi, V. Paramesh, S. R. Kaviya, E. Anuradha, and F. D. Paul Solomon, “3D cell culture systems: Advantages and applications,” J. Cell. Physiol., vol. 230, no. 1, pp. 16–26, 2015. [3] J. W. Haycock, 3D cell culture: a review of current approaches and techniques., vol. 695. 2011. [4] F. Pampaloni, E. G. Reynaud, and E. H. K. Stelzer, “The third dimension bridges the gap between cell culture and live tissue.,” Nat. Rev. Mol. Cell Biol., vol. 8, no. 10, pp. 839–845, 2007. [5] J. Lee, M. J. Cuddihy, and N. A. Kotov, “Three-dimensional cell culture matrices: state of the art.,” Tissue Eng Part B Rev, vol. 14, no. 1, pp. 61–86, 2008. [6] M. Vinci et al., “Advances in establishment and analysis of three-dimensional tumor spheroid-based functional assays for target validation and drug evaluation,” BMC Biol., vol. 10, no. 1, p. 29, 2012. [7] B. A. Justice, N. A. Badr, and R. A. Felder, “3D cell culture opens new dimensions in cell-based assays,” Drug Discov. Today, vol. 14, no. 1–2, pp. 102–107, 2009. [8] I. Meyvantsson and D. J. Beebe, “Cell culture models in microfluidic systems.,” Annu. Rev. Anal. Chem., vol. 1, pp. 423–449, 2008. [9] E. W. K. Young and D. J. Beebe, “Fundamentals of microfluidic cell culture in controlled microenvironments,” Chem Soc Rev, vol. 39, no. 3, pp. 1036–1048, 2010. [10] D. J. Beebe, G. a Mensing, and G. M. Walker, “Physics and applications of microfluidics in biology.,” Annu. Rev. Biomed. Eng., vol. 4, pp. 261–286, 2002. [11] J. El-Ali, P. K. Sorger, and K. F. Jensen, “Cells on chips.,” Nature, vol. 442, no. 7101, pp. 403–411, 2006. [12] J. Guck et al., “Optical deformability as an inherent cell marker for testing malignant transformation and metastatic competence,” Biophys J, vol. 88, no. 5, pp. 3689–3698, 2005. [13] S. K?ster et al., “Drop-based microfluidic devices for encapsulation of single cells.,” Lab Chip, vol. 8, no. 7, pp. 1110–1115, 2008. [14] H. Andersson and A. Van den Berg, “Microfluidic devices for cellomics: A review,” Sensors Actuators, B Chem., vol. 92, no. 3, pp. 315–325, 2003. [15] M. W. Tibbitt and K. S. Anseth, “Hydrogels as extracellular matrix mimics for 3D cell culture,” Biotechnol. Bioeng., vol. 103, no. 4, pp. 655–663, 2009. [16] J. P. Vacanti and R. Langer, “Tissue engineering: the design and fabrication of living replacement devices for surgical reconstruction and transplantation.,” Lancet, vol. 354, p. SI32-I34, 1999. [17] G. S. D. Hetal Patel, Minal Bonde, “Biodegradable polymer scaffolds for tissue engineering,” Trends Biomater. Artif. Organs, vol. 25, no. 1, pp. 20–29, 2011. [18] L. G. Griffith and M. A. Swartz, “Capturing complex 3D tissue physiology in vitro.,” Nat. Rev. Mol. cell Biol., vol. 7, no. 3, pp. 211–24, 2006. [19] D. J. Tobin, “Scaffolds for Tissue Engineering and 3D Cell Culture,” Methods Mol. Biol., vol. 695, no. 2, pp. 213–227, 2011. [20] J. Naranda et al., “Polyester type polyHIPE scaffolds with an interconnected porous structure for cartilage regeneration,” Sci. Rep., vol. 6, no. February, p. 28695, 2016. [21] B. Dhandayuthapani, Y. Yoshida, T. Maekawa, and D. S. Kumar, “Polymeric scaffolds in tissue engineering application: A review,” Int. J. Polym. Sci., vol. 2011, no. ii, 2011. [22] F. J. O’Brien, “Biomaterials & scaffolds for tissue engineering,” Mater. Today, vol. 14, no. 3, pp. 88–95, 2011. [23] A. L. Paguirigan and D. J. Beebe, “Microfluidics meet cell biology: Bridging the gap by validation and application of microscale techniques for cell biological assays,” BioEssays, vol. 30, no. 9, pp. 811–821, Sep. 2008. [24] F.-Q. Nie, M. Yamada, J. Kobayashi, M. Yamato, A. Kikuchi, and T. Okano, “On-chip cell migration assay using microfluidic channels.,” Biomaterials, vol. 28, no. 27, pp. 4017–4022, 2007. [25] A. Valster, N. L. Tran, M. Nakada, M. E. Berens, A. Y. Chan, and M. Symons, “Cell migration and invasion assays,” Methods, vol. 37, no. 2, pp. 208–215, 2005. [26] C. R. Justus, N. Leffler, M. Ruiz-Echevarria, and L. V Yang, “In vitro cell migration and invasion assays.,” J. Vis. Exp., vol. 752, no. 88, p. e51046, 2014. [27] N. Kramer et al., “In vitro cell migration and invasion assays.,” Mutat Res, vol. 752, no. 1, pp. 10–24, 2013. [28] J. W. Hong, V. Studer, G. Hang, W. F. Anderson, and S. R. Quake, “A nanoliter-scale nucleic acid processor with parallel architecture.,” Nat. Biotechnol., vol. 22, no. 4, pp. 435–439, 2004. [29] J. Q. Boedicker, L. Li, T. R. Kline, and R. F. Ismagilov, “Detecting bacteria and determining their susceptibility to antibiotics by stochastic confinement in nanoliter droplets using plug-based microfluidics.,” Lab Chip, vol. 8, no. 8, pp. 1265–1272, 2008. [30] G. Velve-Casquillas, M. Le Berre, M. Piel, and P. T. Tran, “Microfluidic tools for cell biological research,” Nano Today, vol. 5, no. 1. pp. 28–47, 2010. [31] C. R. Terenna et al., “Physical Mechanisms Redirecting Cell Polarity and Cell Shape in Fission Yeast,” Curr. Biol., vol. 18, no. 22, pp. 1748–1753, Nov. 2008. [32] G. Faure-andré, “Regulation of Dendritic Cell Migration by CD74, the MHC Class II–Associated Invariant Chain,” Science (80-. )., vol. 1705, no. December, 2008. [33] S. M. McFaul, B. K. Lin, and H. Ma, “Cell separation based on size and deformability using microfluidic funnel ratchets,” Lab Chip, vol. 12, no. 13, pp. 2369–2376, 2012. [34] S. C. Hur, N. K. Henderson-MacLennan, E. R. B. McCabe, and D. Di Carlo, “Deformability-based cell classification and enrichment using inertial microfluidics.,” Lab Chip, vol. 11, no. 5, pp. 912–920, 2011. [35] H. W. Hou, Q. S. Li, G. Y. H. Lee, A. P. Kumar, C. N. Ong, and C. T. Lim, “Deformability study of breast cancer cells using microfluidics,” Biomed. Microdevices, vol. 11, no. 3, pp. 557–564, 2009.
相關(guān)產(chǎn)品： 1、細(xì)胞力產(chǎn)生和力傳遞綜合測量分析系統(tǒng) 2、各種生物力學(xué)設(shè)備
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