Microfluidic characterization of macromolecular liquid-liquid phase separation.
Microfluidic characterization of macromolecular liquid-liquid phase separation.
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DOI：
10.1039/d0lc00613k
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发表时间：
2020-11-10
期刊：
影响因子：
6.1
通讯作者：
Heymann M
中科院分区：
文献类型：
--
作者：
Bremer A;Mittag T;Heymann M
液-液相分离在细胞的区室化中起着重要作用。要了解生物大分子中相分离是如何编码的，需要对其相行为进行定量映射。鉴于此类实验需要大量感兴趣的生物分子，这些努力一直落后于最近生物学见解的广度。在此，我们提出了一种微流控相芯片，使饱和浓度的测量超过至少三个数量级的广谱的生物分子和溶液条件。相位芯片由五个单元组成，每个单元由二十个单独的样品室组成，可以同时测量五种样品条件。分析物通过水的蒸发缓慢浓缩，水被油取代，直到样品经历相分离成稀相和密相。我们表明，相芯片降低了所需的样品量的98%，同时提供六倍更好的统计相比，标准的手动实验，涉及离心分离的稀相和浓相。我们进一步表明，在芯片中测得的饱和浓度与以前报道的各种生物分子的数据一致。同时，可以通过显微镜监测密相形态的时间依赖性变化和潜在的路径外过程，包括聚集。总之，相位芯片适合于通过以低蛋白质成本确定大量饱和浓度来探索序列与双节点的关系。
Liquid-liquid phase separation plays important roles in the compartmentalization of cells. Developing an understanding of how phase separation is encoded in biomacromolecules requires quantitative mapping of their phase behavior. Given that such experiments require large quantities of the biomolecule of interest, these efforts have been lagging behind the recent breadth of biological insights. Herein, we present a microfluidic phase chip that enables the measurement of saturation concentrations over at least three orders of magnitude for a broad spectrum of biomolecules and solution conditions. The phase chip consists of five units, each made of twenty individual sample chambers to allow the measurement of five sample conditions simultaneously. The analytes are slowly concentrated via evaporation of water, which is replaced by oil, until the sample undergoes phase separation into a dilute and dense phase. We show that the phase chip lowers the required sample quantity by 98% while offering six-fold better statistics in comparison to standard manual experiments that involve centrifugal separation of dilute and dense phases. We further show that the saturation concentrations measured in chips are in agreement with previously reported data for a variety of biomolecules. Concomitantly, time-dependent changes of the dense phase morphology and potential off-pathway processes, including aggregation, can be monitored microscopically. In summary, the phase chip is suited to exploring sequence-to-binodal relationships by enabling the determination of a large number of saturation concentrations at low protein cost.
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在细胞周期中进行逐步估计。
影响因子：
2.3
作者：
Boeynaems S;Tompa P;Van Den Bosch L
通讯作者：
Van Den Bosch L
肌萎缩性侧索硬化和额颞痴呆的TIA1突变促进相位分离并改变应激颗粒动力学。
影响因子：
16.2
作者：
Mackenzie IR;Nicholson AM;Sarkar M;Messing J;Purice MD;Pottier C;Annu K;Baker M;Perkerson RB;Kurti A;Matchett BJ;Mittag T;Temirov J;Hsiung GR;Krieger C;Murray ME;Kato M;Fryer JD;Petrucelli L;Zinman L;Weintraub S;Mesulam M;Keith J;Zivkovic SA;Hirsch-Reinshagen V;Roos RP;Züchner S;Graff-Radford NR;Petersen RC;Caselli RJ;Wszolek ZK;Finger E;Lippa C;Lacomis D;Stewart H;Dickson DW;Kim HJ;Rogaeva E;Bigio E;Boylan KB;Taylor JP;Rademakers R
通讯作者：
Rademakers R
影响因子：
8.6
作者：
Lin, Yi-Hsuan;Forman-Kay, Julie D.;Chan, Hue Sun
通讯作者：
Chan, Hue Sun
影响因子：
3.3
作者：
Das, Suman;Eisen, Adam;Chan, Hue Sun
通讯作者：
Chan, Hue Sun
影响因子：
56.9
作者：
Franzmann, Titus M.;Jahnel, Marcus;Alberti, Simon
通讯作者：
Alberti, Simon