Name: a small volume bioassay to assess bacterial/phytoplankton co-culture using water-pulse-amplitude-modulated (water-pam) fluorometry
Uploaded: 2015-03-11T13:00:00
Description: Watch this Scientific Journal Video about A Small Volume Bioassay to Assess Bacterial/Phytoplankton Co-culture Using WATER-Pulse-Amplitude-Modulated WATER-PAM Fluorometry at JoVE.com

This work was supported by Natural Sciences and Engineering Research Council of Canada (grant 402105), Canadian Foundation for Innovation (grant 129087) and Alberta Education and Training (grant AAETRCP-12-026-SEG) to RJC.

1.计算实验装置<ol><li>计算藻类和/或所需要的，这将需要为整个实验通过使用等式1控制细菌培养物的体积： <img alt="式（1）" src="/files/ftp_upload/52455/52455equation1.1.jpg" /> 其中，y等于每天和z所需的控制的数目等于的天数。 </li><li>计算是通过使用等式2所需要的共培养物的实验是藻类和/或细菌培养物的体积： <img alt="公式2" src="/files/ftp_upload/52455/52455equation1.2.jpg" /> 注：这是可能取代“...

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藻类生长在一个小型化的格式。 藻培养物在微量滴定板1毫升培养物体积的小型化允许一个实验中的复制增加。以确保藻是在整个实验的健康是非常重要的;执行一个生长曲线（ 图2），使用微量滴定板形式，以评估各种藻类介质，以确保满足所述藻类的营养需求。此外，为了优化昼夜周期（亮和暗期）和温度可能是重要的。适当的优化对于给定的藻类可以允许用于?...

浮游植物生理学历来研究中尺度实验范围从20毫升锥形瓶中，以5升大玻璃瓶中1-7。这个试验规模需要二次抽样实验监测，作为牺牲重复样品对每个时间点创建一个无法控制的实验装置。 的能力，同时使用相同的昼夜培养箱空间通过小型化的实验量为藻类生理学实验将减少或消除二次抽样和伪复制的限制从大量增加的独立实验的数目。微孔板格式已经开发出用1毫升培养体积的实验操作藻可变的条件藻类生物测定。这种小规模的试验体积允许要增加重复的次数，增加实验重复性由于重复样品之间降低的变异性实验中，并且允许真复制，同时保持试验对照（ 即无菌藻类培养物）140天（ 图2）12。 这种微量滴定板形式很容易适应于多种实验问题，如：没有细菌具有与其藻宿主共生，中性或致病相互作用？是另外一种化合物刺激或有毒的藻类？这些和其它问题，可在使用这种新格式9-11快速高通量的方式加以解决。 48孔微量滴定培养皿允许每个1毫升井是一个独立的实验装置所采样的单个时间点。各种参数可以从这个1毫升体积包括被采样，但不限于：使用水-脉冲幅度调制（WATER-PAM）荧光（参见材料和设备表 ）1叶绿素荧光和光化学参数3。WATER-PAM荧光是一种快速，非侵入性的技术，可用于监测与藻类13进行的实验。它允许光合效率和PSII健康从小培养体积测量（150 - 300μl的文化在培养基中稀释到2 - 4毫升体积WATER-PAM）14,15。除了水-PAM荧光，这种设置可以用来测量各种其它参数，包括但不限于：显微镜可视化附着到的藻细胞和变化，藻细胞形态的细菌;细菌菌落形成单位（CFU）计数;和流式细胞仪对藻类细胞计数和识别的亚群。

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			<td height="21" style="height:21px;width:413px;">Name of Material/ Equipment</td>
			<td style="width:328px;">Company</td>
			<td style="width:331px;">Catalog Number</td>
			<td style="width:163px;">Comments/Description</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">10 cu. ft. Diurnal Incubator (6012-1)</td>
			<td>Caron Corporate</td>
			<td>112310-6012-1-11</td>
			<td><a href="http://www.caronproducts.com">www.caronproducts.com</a></td>
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		<tr height="23">
			<td height="23" style="height:23px;">Nunc EasYFlask 25cm2, Vent/Close Cap, 7mL working volume, 200/Cs&#160;</td>
			<td>Thermo Fisher Scientific</td>
			<td>N156340</td>
			<td><a href="http://www.fishersci.ca">www.fishersci.ca</a></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Multiwell TC Plates &#8211; 48 Well</td>
			<td>BD Biosciences Discovery Labware</td>
			<td>353078</td>
			<td><a href="http://www.bdbiosciences.com/">www.bdbiosciences.com</a></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">P1000 Gilson The Pipetting Standard&#8212;Gilson&#39;s Pipetman</td>
			<td>Mandel Scientific Company Inc.</td>
			<td>GF-F123602</td>
			<td><a href="http://www.mandel.ca">www.mandel.ca</a></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">P10mL Gilson The Pipetting Standard&#8212;Gilson&#39;s Pipetman</td>
			<td>Mandel Scientific Company Inc.</td>
			<td>GF-F161201</td>
			<td><a href="http://www.mandel.ca">www.mandel.ca</a></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Wide Orifice Tips nonsterile [100&#8211;1250 &#181;L]</td>
			<td>VWR International</td>
			<td>89079-468</td>
			<td><a href="http://www.ca.vwr.com">www.ca.vwr.com</a></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Ultrafine Tips nonsterile [100&#8211;1250 &#181;L]</td>
			<td>VWR International</td>
			<td>89079-470</td>
			<td><a href="http://www.ca.vwr.com">www.ca.vwr.com</a></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Finntip 10mL [Vol: 1-10mL]</td>
			<td>Thermo Fisher Scientific</td>
			<td>9402151</td>
			<td><a href="http://www.fishersci.ca">www.fishersci.ca</a></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">WATER-Pulse Amplitude Modulation (Water-ED)</td>
			<td>Heinz Walz GmbH, Effeltrich, Germany</td>
			<td>EDEE0232</td>
			<td><a href="http://www.walz.com">www.walz.com</a></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">15 mm diameter quartz glass cuvette (WATER-K)</td>
			<td>Caron Corporate</td>
			<td></td>
			<td><a href="http://www.caronproducts.com">www.caronproducts.com</a></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Sodium&#160;Chloride&#160;(Crystalline/Certified ACS), Fisher Chemical</td>
			<td>Thermo Fisher Scientific</td>
			<td>Thermo Fisher Scientific</td>
			<td><a href="http://www.fishersci.ca">www.fishersci.ca</a></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">BD Difco Marine Broth 2216</td>
			<td>BD Biosciences Discovery Labware</td>
			<td>BD Biosciences Discovery Labware</td>
			<td><a href="http://www.bdbiosciences.com">www.bdbiosciences.com</a></td>
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		<tr height="20">
			<td height="20" style="height:20px;">BD Bacto Agar</td>
			<td>BD Biosciences Discovery Labware</td>
			<td>BD Biosciences Discovery Labware</td>
			<td><a href="http://www.bdbiosciences.com">www.bdbiosciences.com</a></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">L1 Medium Kit, 50L</td>
			<td>NCMA [National Center for Marine Algae and Microbiota</td>
			<td>NCMA [National Center for Marine Algae and Microbiota</td>
			<td><a href="http://www.ncma.bigelow.org">www.ncma.bigelow.org</a></td>
		</tr>
	</tbody>
</table>

a small volume bioassay to assess bacterial/phytoplankton co-culture using water-pulse-amplitude-modulated (water-pam) fluorometry

常规方法实验操作微藻雇用大量培养物（20毫升至5升），以使培养能在整个实验1-7进行二次取样。大容量二次抽样可能会产生问题，原因如下：1）它引起的变化的总体积与表面积：在实验过程中培养的体积比; 2）伪复制（ 即复制从相同的待遇烧瓶8个样本）经常采用而不是真正的复制（ 即从重复治疗抽样）; 3）在实验的持续时间是有限的总体积;和4）纯性培养物或通常的细菌微生物是困难的，因为污染子采样期间通常发生在长期实验来维持。 使用微量滴定板的使1毫升培养体积要用于每个复制，与内多达48个独立的处理一12.65 X 8.5×2.2厘米板，从而降低了实验体积并允许广泛的复制子采样未经任何处理。此外，这种技术可以被修改以适应多种实验格式，包括：细菌藻共培养，藻类的生理测试，以及毒素筛选9-11。各孔用藻类，细菌和/或共培养物可以被采样为众多的实验室程序，包括但不限于：水 - 脉冲幅度调制（WATER-PAM）荧光，显微镜检查，细菌菌落形成单位（cfu）的计数和流式细胞术。微量滴定板格式和水的PAM荧光的组合允许光化学产量与样品中，高再现性之间的低变异性等光化学参数的多个快速测量，并避免了子采样一个大玻璃瓶或锥形瓶中在实验的过程中的许多陷阱。

WATER-PAM荧光读数。 WATER-脉冲幅度调制（PAM）荧光是一种快速而有效的方法来确定荧光（叶绿素含量代理）和藻类培养光合产量（PSII健康）。在PAM WINCONTROL软件生成原始数据值的表格（下面是对暗适于藻类样品的基本参数）： f 0的=荧光暗适应的细胞的 F M =最大荧光饱和发光二极管后（LED）的脉冲?...

Watch this Scientific Journal Video about A Small Volume Bioassay to Assess Bacterial/Phytoplankton Co-culture Using WATER-Pulse-Amplitude-Modulated WATER-PAM Fluorometry at JoVE.com

A Small Volume Bioassay to Assess Bacterial/Phytoplankton Co-culture Using WATER-Pulse-Amplitude-Modulated WATER-PAM Fluorometry

The goal of this procedure is to demonstrate the reproducibility and adaptability of using a microtiter plate format for microalgal screening. This rapid screen combines WATER-Pulse-Amplitude-Modulated (WATER-PAM) fluorometry to measure photosynthetic yield as an indicator of Photosystem II (PSII) health with small volume bacterial-algal co-cultures.

小体积的生物测定评估细菌/浮游植物共培养使用水脉冲幅度调制（WATER-PAM）荧光

Conventional methods for experimental manipulation of microalgae have employed large volumes of culture (20 ml to 5 L), so that the culture can be ...

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A Small Volume Bioassay to Assess Bacterial/Phytoplankton Co-culture Using WATER-Pulse-Amplitude-Modulated (WATER-PAM) Fluorometry

Optimized Negative Staining: a High-throughput Protocol for Examining Small and Asymmetric Protein Structure by Electron Microscopy

优化负染：通过电子显微镜检查小和不对称蛋白结构的高通量协议

Structural determination of proteins is rather challenging for proteins with molecular masses between 40 - 200 kDa. Considering that more than half of ...

科研

环境科学

A Small Volume Procedure for Viral Concentration from Water

小体积的方法从水中病毒浓度

Small-scale concentration of viruses (sample volumes 1-10 L, here simulated with spiked 100 ml water samples) is an efficient, cost-effective way to ...

A Whole Cell Bioreporter Approach to Assess Transport and Bioavailability of Organic Contaminants in Water Unsaturated Systems

全细胞Bioreporter方法​​评估交通有机污染物的生物利用度和水饱和系统

Bioavailability of contaminants is a prerequisite for their effective biodegradation in soil. The average bulk concentration of a contaminant, however, is ...

A Fish-feeding Laboratory Bioassay to Assess the Antipredatory Activity of Secondary Metabolites from the Tissues of Marine Organisms

了鱼喂食实验室生物测定，从海洋生物的组织评估次生代谢产物的Antipredatory活动

Marine chemical ecology is a young discipline, having emerged from the collaboration of natural products chemists and marine ecologists in the 1980s with ...

A Simple Method for Automated Solid Phase Extraction of Water Samples for Immunological Analysis of Small Pollutants

对水样的自动固相萃取小污染物的免疫学分析的简单方法

A new method for solid phase extraction (SPE) of environmental water samples is proposed. The developed prototype is cost-efficient and user friendly, and ...

Extracting Metrics for Three-dimensional Root Systems: Volume and Surface Analysis from In-soil X-ray Computed Tomography Data

从在土X射线计算机断层扫描数据量和表面分析：提取指标三维根系

Plant roots play a critical role in plant-soil-microbe interactions that occur in the rhizosphere, as well as processes with important implications to ...

How to Create Conditioned Taste Aversion for Grazing Ground Covers in Woody Crops with Small Ruminants

如何创建条件性味觉厌恶放牧在地面木本作物与小反刍动物封面

Conditioned taste aversion (CTA) is a learning behavior process where animals are trained to reject certain feed after gastrointestinal discomfort has ...

The Benthic Exchange of O2, N2 and Dissolved Nutrients Using Small Core Incubations

The Benthic Exchange of O2, N2 and Dissolved Nutrients Using Small Core Incubations

Ø的底栖交易所<sub&gt; 2</sub&gt;，N<sub&gt; 2</sub&gt;和营养盐使用小的核心孵化

The measurement of sediment-water exchange of gases and solutes in aquatic sediments provides data valuable for understanding the role of sediments in ...

Measuring the Flight Ability of the Ambrosia Beetle, Platypus Quercivorus (Murayama), Using a Low-Cost, Small, and Easily Constructed Flight Mill

Measuring the Flight Ability of the Ambrosia Beetle, Platypus Quercivorus Murayama, Using a Low-Cost, Small, and Easily Constructed Flight Mill

利用低成本、小型、易于建造的Quercivorus测量豚草甲虫、鸭嘴兽的飞行能力 (村山富市)

The ambrosia beetle, Platypus quercivorus (Murayama), is the vector of a fungal pathogen that causes mass mortality of Fagaceae trees (Japanese oak wilt). ...