Extracellular matrix micropatterning technology for whole cell cryogenic electron microscopy studies

被引：22

作者：

Engel, Leeya ^{[1
]}

Gaietta, Guido ^{[2
]}

Dow, Liam P. ^{[1
,3
]}

Swif, Mark F. ^{[2
]}

Pardon, Gaspard ^{[1
]}

Volkmann, Niels ^{[2
]}

Weis, William, I ^{[6
,7
]}

Hanein, Dorit ^{[2
]}

Pruitt, Beth L. ^{[1
,3
,4
,5
]}

机构：

[1] Stanford Univ, Dept Bioengn, Stanford, CA 94305 USA

[2] Sanford Burnham Prebys Med Discovery Inst, Immun & Pathogenesis Program, La Jolla, CA USA

[3] Univ Calif Santa Barbara, Biomol Sci & Engn Program, Santa Barbara, CA 93106 USA

[4] Univ Calif Santa Barbara, Dept Mech Engn, Santa Barbara, CA 93106 USA

[5] Univ Calif Santa Barbara, Dept Mol Cellular & Dev Biol, Santa Barbara, CA 93106 USA

[6] Stanford Univ, Sch Med, Dept Struct Biol, Stanford, CA 94305 USA

[7] Stanford Univ, Sch Med, Dept Mol & Cellular Physiol, Stanford, CA 94305 USA

来源：

JOURNAL OF MICROMECHANICS AND MICROENGINEERING | 2019年 / 29卷 / 11期

基金：

美国国家科学基金会; 美国国家卫生研究院;

关键词：

micropatterning; cryo-EM; cryo-ET; mechanobiology; transmission electron microscopy; bioengineering; electron cryotomography; CRYOELECTRON TOMOGRAPHY; MOLECULAR SOCIOLOGY; SOFT LITHOGRAPHY; STEM-CELL; SHAPE; ARCHITECTURE; GEOMETRY; DIFFERENTIATION; FLUORESCENCE; ORGANIZATION;

D O I：

10.1088/1361-6439/ab419a

中图分类号：

TM [电工技术]; TN [电子技术、通信技术];

学科分类号：

0808 ; 0809 ;

摘要：

Cryogenic electron tomography is the highest resolution tool available for structural analysis of macromolecular organization inside cells. Micropatterning of extracellular matrix (ECM) proteins is an established in vitro cell culture technique used to control cell shape. Recent traction force microscopy studies have shown correlation between cell morphology and the regulation of force transmission. However, it remains unknown how cells sustain increased strain energy states and localized stresses at the supramolecular level. Here, we report a technology to enable direct observation of mesoscale organization in epithelial cells under morphological modulation, using a maskless protein photopatterning method (PRIMO) to confine cells to ECM micropatterns on electron microscopy substrates. These micropatterned cell culture substrates can be used in mechanobiology research to correlate changes in manometer-scale organization at cell-cell and cell-ECM contacts to strain energy states and traction stress distribution in the cell.

引用

页数：9

共 49 条

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