Continuous and long-term volume measurements with a commercial Coulter counter.
Continuous and long-term volume measurements with a commercial Coulter counter.
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DOI：
10.1371/journal.pone.0029866
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Manalis SR
中科院分区：
文献类型：
--
作者：
Bryan AK;Engler A;Gulati A;Manalis SR
我们展示了一种方法来提高商用库尔特计数器的时间分辨率，并实现连续和长期的细胞大小测量，以进行生长速度分析，这对于了解基本的细胞过程至关重要，如细胞大小调节和细胞周期进展。我们对商用库尔特计数器的简单改装在样品室内创造了可控的细胞培养条件，并将温度控制与必要的适应相结合，以实现数小时的测量稳定性。我们还编写了定制软件，在此详细介绍，用于分析通过这种连续方法或标准定期采样收集的仪器数据文件。我们使用连续方法来测量酵母在长时间停滞期间在G1中的生长速度，并在不同的样本中，测量停滞和增殖细胞中细胞大小和细胞周期位置对生长速度的依赖关系。我们还用高时间分辨率量化了小鼠淋巴母细胞培养对药物治疗的反应。这种方法提供了一种适用于多种细胞类型的连续测量细胞大小的技术，并极大地扩展了库尔特计数器可用的分析工具集。
We demonstrate a method to enhance the time resolution of a commercial Coulter counter and enable continuous and long-term cell size measurements for growth rate analyses essential to understanding basic cellular processes, such as cell size regulation and cell cycle progression. Our simple modifications to a commercial Coulter counter create controllable cell culture conditions within the sample compartment and combine temperature control with necessary adaptations to achieve measurement stability over several hours. We also wrote custom software, detailed here, to analyze instrument data files collected by either this continuous method or standard, periodic sampling. We use the continuous method to measure the growth rate of yeast in G1 during a prolonged arrest and, in different samples, the dependency of growth rate on cell size and cell cycle position in arrested and proliferating cells. We also quantify with high time resolution the response of mouse lymphoblast cell culture to drug treatment. This method provides a technique for continuous measurement of cell size that is applicable to a large variety of cell types and greatly expands the set of analysis tools available for the Coulter counter.
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影响因子：
48
作者：
Godin, Michel;Delgado, Francisco Feijo;Son, Sungmin;Grover, William H.;Bryan, Andrea K.;Tzur, Amit;Jorgensen, Paul;Payer, Kris;Grossman, Alan D.;Kirschner, Marc W.;Manalis, Scott R.
通讯作者：
Manalis, Scott R.
影响因子：
3.2
作者：
HARTWELL, LH
通讯作者：
HARTWELL, LH
影响因子：
2.4
作者：
BERGE, LI;JOSSANG, T;FEDER, J
通讯作者：
FEDER, J
影响因子：
3.7
作者：
Tzur A;Moore JK;Jorgensen P;Shapiro HM;Kirschner MW
通讯作者：
Kirschner MW
影响因子：
64.8
作者：
Bishop, AC;Ubersax, JA;Shokat, KM
通讯作者：
Shokat, KM