Theory and practice of using cell strainers to sort Caenorhabditis elegans by size.
Theory and practice of using cell strainers to sort Caenorhabditis elegans by size.
复制标题
DOI：
10.1371/journal.pone.0280999
复制
发表时间：
2023
期刊：
影响因子：
3.7
通讯作者：
Lockery, Shawn R.
中科院分区：
文献类型：
--
作者：
Lanier, Vincent J.;White, Amanda M.;Faumont, Serge;Lockery, Shawn R.
秀丽线虫是一种广泛应用于基础研究、转译研究和工业研究的模式生物。线虫的发育有5个不同的形态阶段，包括4个幼虫期和成虫期。不同的阶段在大小、基因表达、生理和行为等方面都不同。对特定发育阶段的丰富往往是实验设计的第一步。当需要数百条虫子时，标准的繁殖方法是在固定的时间内培育同步孵化的幼虫种群，或者根据大小对混合种群的虫子进行分类。目前的大小排序方法比同步方法具有更高的吞吐量，并避免了其使用苛刻的化学物质。然而，这些大小分选方法目前需要昂贵的仪器或定制的微流控设备，这两种设备对大多数线虫实验室来说都是不可用的。因此，需要廉价、可访问的分类策略。我们研究了使用低成本、商业可用的细胞过滤器来按大小过滤线虫。我们发现，对于三种不同目次的细胞过滤器，过滤后恢复的概率是身体大小的函数，可以用Logistic函数很好地描述。应用这些函数预测过滤结果显示，细胞过滤器对蠕虫的过滤并不理想，但却提高了过滤结果。此外，我们发现，使用一对具有不同网目大小的过滤器的串联过滤可以用于特定幼虫阶段的浓缩，其纯度接近最广泛使用的同步浓缩方法。细胞过滤器法的产量高达每分钟14,000条蠕虫，大大超过了其他富集法。我们的结论是，通过细胞过滤器进行大小排序是对现有方法的有益补充，用于丰富线虫特定发育阶段。
The nematode Caenorhabditis elegans is a model organism widely used in basic, translational, and industrial research. C. elegans development is characterized by five morphologically distinct stages, including four larval stages and the adult stage. Stages differ in a variety of aspects including size, gene expression, physiology, and behavior. Enrichment for a particular developmental stage is often the first step in experimental design. When many hundreds of worms are required, the standard methods of enrichment are to grow a synchronized population of hatchlings for a fixed time, or to sort a mixed population of worms according to size. Current size-sorting methods have higher throughput than synchronization and avoid its use of harsh chemicals. However, these size-sorting methods currently require expensive instrumentation or custom microfluidic devices, both of which are unavailable to the majority C. elegans laboratories. Accordingly, there is a need for inexpensive, accessible sorting strategies. We investigated the use of low-cost, commercially available cell strainers to filter C. elegans by size. We found that the probability of recovery after filtration as a function of body size for cell strainers of three different mesh sizes is well described by logistic functions. Application of these functions to predict filtration outcomes revealed non-ideal properties of filtration of worms by cell strainers that nevertheless enhanced filtration outcomes. Further, we found that serial filtration using a pair of strainers that have different mesh sizes can be used to enrich for particular larval stages with a purity close to that of synchronization, the most widely used enrichment method. Throughput of the cell strainer method, up to 14,000 worms per minute, greatly exceeds that of other enrichment methods. We conclude that size sorting by cell strainers is a useful addition to the array of existing methods for enrichment of particular developmental stages in C. elegans.
登录
查看更多内容
影响因子：
6.1
作者：
Han, Bicheng;Kim, Daeyeon;Shin, Jennifer H.
通讯作者：
Shin, Jennifer H.
影响因子：
3.5
作者：
LUDWICK, JC;HENDERSO.PL
通讯作者：
HENDERSO.PL
影响因子：
6.1
作者：
Ai X;Zhuo W;Liang Q;McGrath PT;Lu H
通讯作者：
Lu H
影响因子：
3.4
作者：
Mutwakil, MHAZ;Steele, TJG;dePomerai, DI
通讯作者：
dePomerai, DI
使用微流控阻抗细胞术测量线虫并确定其发育阶段
影响因子：
8.4
作者：
Zhu, Zhen;Chen, Weijie;Pan, Dejing
通讯作者：
Pan, Dejing