High-throughput co-encapsulation of self-ordered cell trains: cell pair interactions in microdroplets
High-throughput co-encapsulation of self-ordered cell trains: cell pair interactions in microdroplets
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DOI：
10.1039/c3ra43624a
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发表时间：
2013-01-01
期刊：
影响因子：
3.9
通讯作者：
Edd, Jon F.
中科院分区：
文献类型：
--
作者：
Lagus, Todd P.;Edd, Jon F.
液滴微流体是微流体的一个蓬勃发展的子领域，它为细胞分析增加了限制的好处，包括信号积累和隔离。然而，控制每个液滴的细胞数量已被限制为使用泊松（随机）包封以获得最高通量。包含一个且仅一个细胞的液滴的泊松概率被限制为36.8%，并且在液滴中配对两种不同细胞类型的概率被限制为13.5%。将液滴微流体与惯性微流体相结合，我们提出了一种以6 kHz的速率有效地将细胞对共封装在液滴中的装置。我们证明了颗粒共包封，其中64%的液滴包含正确的一对一配对，这代表了泊松共包封的近五倍改善。我们还应用该设备封装两个单独的莱茵衣原体菌株。C. Reinhardtii是一种单细胞微藻，可用作模式生物、重组蛋白源和多种生物燃料的潜在来源。在通过氮饥饿和热诱导鞭毛损失诱导配子发生后，我们共封装单独的交配型正（mt+）和交配型负（mt-）C。reinhardtii细胞液滴。在这里，29%的液滴含有一个且仅一个每种交配类型的细胞，比泊松共包封概率13%提高了两倍。爆燃后约1小时，配子恢复鞭毛运动和交配能力的液滴内。将交配的受精卵以乳液形式储存而不补充营养。17天后，接合孢子和一些未交配的配子仍然存活。当乳状液被打破并接种在全营养琼脂上时，接合孢子萌发，四分体孵化，然后进行有丝分裂。除了藻类，该设备有可能对各种细胞类型进行有限的相互作用研究。
Droplet microfluidics is a booming sub-field of microfluidics that adds the benefits of confinement, including signal accumulation and isolation, to cell analysis. However, controlling the number of cells per droplet has been limited to using Poisson (random) encapsulation for the highest throughputs. The Poisson probability of a droplet containing one and only one cell is limited to 36.8%, and the probability of pairing two distinct cell types in a droplet is limited to 13.5%. Combining droplet microfluidics with inertial microfluidics, we present a device which efficiently co-encapsulates cell pairs in droplets at rates on the order of 6 kHz. We demonstrate particle co-encapsulation where 64% of droplets contained the correct one-to-one pairing, representing a nearly fivefold improvement to Poisson co-encapsulation. We also apply the device to encapsulate two separate strains of Chlamydomonas reinhardtii. C. reinhardtii is a single-cell microalgae with applications as a model organism, recombinant protein source, and potential source of multiple biofuels. After inducing gametogenesis by nitrogen starvation and thermally inducing flagella loss, we co-encapsulate separate mating-type plus (mt+) and mating-type minus (mt-) C. reinhardtii cells in droplets. Here, 29% of droplets contained one and only one cell of each mating type, over a twofold improvement to the Poisson co-encapsulation probability of 13%. Approximately one hour following deflagellation, gametes regained flagellar motility and mating ability within the droplets. The mated zygotes were stored in emulsion form without nutrient replenishment. After 17 days, both zygospores and, remarkably, some unmated gametes remained viable. When the emulsion was broken and plated on full-nutrient agar, zygospore germination, tetrad hatching, and then mitosis followed. In addition to algae, the device has the potential for confined interaction studies for a variety of cell types.