Name: a small-scale setup for algal toxicity testing of nanomaterials and other difficult substances
Uploaded: 2020-10-10T14:00:00
Description: Watch this Scientific Journal Video about A Small-Scale Setup for Algal Toxicity Testing of Nanomaterials and Other Difficult Substances at JoVE.com

这项研究由S前-纳米安全测试先进工具资助，根据Horizon 2020研究和创新计划授予协议760813。

1. 莱维特设置的描述
<ol><li>使用20 mL闪烁玻璃瓶（图1，插入1），允许光线穿透。或者，可以使用轻的可塑胶瓶。使用光度计量化光强。</li>
	<li>在测试开始时至少使用 4 mL 测试悬浮液，以便对生物量进行定量，并在孵化期间和之后对纳米材料进行表征/定量（图 1，插入 2）。</li>
	<li>将 20 mL 闪烁小瓶与盖（图1，插入 3）配合，其...

<ol>
	<li>European Chemicals Agency. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Guidance+on+Registration.">Guidance on Registration.</a> European Chemicals Agency. , (2016).</li><li>Organisation for Economic Cooperation and Development. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Guidance+Document+on+Aquatic+Toxicity+Testing+of+Difficult+Substances+and+Mixtures.">Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures.</a> Organisation for Economic Cooperation and Development. , (2019).</li><li>Blaise, C., Legault, R., Bermingham, N., Van Coillie, R., Vasseur, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+microplate+algal+assay+technique+for+aquatic+toxicity+assessment.">A simple microplate algal assay technique for aquatic toxicity assessment.</a> Toxicity Assessment. 1 (3), 261-281 (1986).</li><li>Hjorth, R., Sorensen, S. N., Olsson, M. E., Baun, A., Hartmann, N. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+certain+shade+of+green:+can+algal+pigments+reveal+shading+effects+of+nanoparticles.">A certain shade of green: can algal pigments reveal shading effects of nanoparticles.</a> Integrated Environmental Assessment and Management. 12 (1), 200-202 (2016).</li><li>Chen, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Algae+response+to+engineered+nanoparticles:+current+understanding{,}+mechanisms+and+implications.">Algae response to engineered nanoparticles: current understanding{,} mechanisms and implications.</a> Environmental Science: Nano. 6 (4), 1026-1042 (2019).</li><li>Arensberg, P., Hemmingsen, V. H., Nyholm, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+miniscale+algal+toxicity+test.">A miniscale algal toxicity test.</a> Chemosphere. 30 (11), 2103-2115 (1995).</li><li>Organisation for Economic Cooperation and Development. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Test+No.+201:+Freshwater+Alga+and+Cyanobacteria,+Growth+Inhibition+Test.">Test No. 201: Freshwater Alga and Cyanobacteria, Growth Inhibition Test.</a> Organisation for Economic Cooperation and Development. , (2011).</li><li>International Organization for Standardization (ISO). <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Water+Quality+-+Fresh+Water+Algal+Growth+Inhibition+Test+with+Unicellular+Green+Algae.">Water Quality - Fresh Water Algal Growth Inhibition Test with Unicellular Green Algae.</a> International Organization for Standardization (ISO). , (2012).</li><li>Halling-S&#248;rensen, B., Nyhohn, N., Baun, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Algal+toxicity+tests+with+volatile+and+hazardous+compounds+in+air-tight+test+flasks+with+CO2+enriched+headspace.">Algal toxicity tests with volatile and hazardous compounds in air-tight test flasks with CO2 enriched headspace.</a> Chemosphere. 32 (8), 1513-1526 (1996).</li><li>Mayer, P., Nyholm, N., Verbruggen, E. M. J., Hermens, J. L. M., Tolls, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Algal+growth+inhibition+test+in+filled,+closed+bottles+for+volatile+and+sorptive+materials.">Algal growth inhibition test in filled, closed bottles for volatile and sorptive materials.</a> Environmental Toxicology and Chemistry. 19 (10), 2551-2556 (2000).</li><li>Ritz, C., Baty, F., Streibig, J. C., Gerhard, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dose-response+analysis+using+R.">Dose-response analysis using R.</a> PloS One. 10 (12), 0146021 (2015).</li><li>Birch, H., Kramer, N. I., Mayer, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Time-resolved+freely+dissolved+concentrations+of+semivolatile+and+hydrophobic+test+chemicals+in+in+vitro+assays-measuring+high+losses+and+crossover+by+headspace+solid-phase+microextraction.">Time-resolved freely dissolved concentrations of semivolatile and hydrophobic test chemicals in in vitro assays-measuring high losses and crossover by headspace solid-phase microextraction.</a> Chemical Research in Toxicology. 32 (9), 1780-1790 (2019).</li><li>Trac, L. N., Schmidt, S. N., Mayer, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Headspace+passive+dosing+of+volatile+hydrophobic+chemicals+-+toxicity+testing+exactly+at+the+saturation+level.">Headspace passive dosing of volatile hydrophobic chemicals - toxicity testing exactly at the saturation level.</a> Chemosphere. 211, 694-700 (2018).</li><li>Eisentraeger, A., Dott, W., Klein, J., Hahn, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+studies+on+algal+toxicity+testing+using+fluorometric+microplate+and+Erlenmeyer+flask+growth-inhibition+assays.">Comparative studies on algal toxicity testing using fluorometric microplate and Erlenmeyer flask growth-inhibition assays.</a> Ecotoxicology and Environmental Safety. 54 (3), 346-354 (2003).</li><li>Paixao, S. M., Silva, L., Fernandes, A., O'Rourke, K., Mendonca, E., Picado, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Performance+of+a+miniaturized+algal+bioassay+in+phytotoxicity+screening.">Performance of a miniaturized algal bioassay in phytotoxicity screening.</a> Ecotoxicology. 17 (3), 165-171 (2008).</li><li>Thellen, C., Blaise, C., Roy, Y., Hickey, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Round-robin+testing+with+the+selenastrum--capricornutum+microplate+toxicity+assay.">Round-robin testing with the selenastrum--capricornutum microplate toxicity assay.</a> Hydrobiologia. 188, 259-268 (1989).</li><li>Nagai, T., Taya, K., Annoh, H., Ishihara, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+a+fluorometric+microplate+algal+toxicity+assay+for+riverine+periphytic+algal+species.">Application of a fluorometric microplate algal toxicity assay for riverine periphytic algal species.</a> Ecotoxicology and Environmental Safety. 94, 37-44 (2013).</li><li>Lee, W. M., An, Y. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+zinc+oxide+and+titanium+dioxide+nanoparticles+on+green+algae+under+visible,+UVA,+and+UVB+irradiations:+no+evidence+of+enhanced+algal+toxicity+under+UV+pre-irradiation.">Effects of zinc oxide and titanium dioxide nanoparticles on green algae under visible, UVA, and UVB irradiations: no evidence of enhanced algal toxicity under UV pre-irradiation.</a> Chemosphere. 91 (4), 536-544 (2013).</li><li>Samei, M., Sarrafzadeh, M. H., Faramarzi, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+impact+of+morphology+and+size+of+zinc+oxide+nanoparticles+on+its+toxicity+to+the+freshwater+microalga,+Raphidocelis+subcapitata.">The impact of morphology and size of zinc oxide nanoparticles on its toxicity to the freshwater microalga, Raphidocelis subcapitata.</a> Environmental Science and Pollution Research. 26 (3), 2409-2420 (2019).</li><li>Neale, P. A., Jaemting, A. K., O'Malley, E., Herrmann, J., Escher, B. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behaviour+of+titanium+dioxide+and+zinc+oxide+nanoparticles+in+the+presence+of+wastewater-derived+organic+matter+and+implications+for+algal+toxicity.">Behaviour of titanium dioxide and zinc oxide nanoparticles in the presence of wastewater-derived organic matter and implications for algal toxicity.</a> Environmental Science: Nano. 2 (1), 86-93 (2015).</li><li>Hartmann, N. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+challenges+of+testing+metal+and+metal+oxide+nanoparticles+in+algal+bioassays:+titanium+dioxide+and+gold+nanoparticles+as+case+studies.">The challenges of testing metal and metal oxide nanoparticles in algal bioassays: titanium dioxide and gold nanoparticles as case studies.</a> Nanotoxicology. 7 (6), 1082-1094 (2013).</li><li>Farkas, J., Booth, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Are+fluorescence-based+chlorophyll+quantification+methods+suitable+for+algae+toxicity+assessment+of+carbon+nanomaterials.">Are fluorescence-based chlorophyll quantification methods suitable for algae toxicity assessment of carbon nanomaterials.</a> Nanotoxicology. 11 (4), 569-577 (2017).</li><li>Handy, R. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Practical+considerations+for+conducting+ecotoxicity+test+methods+with+manufactured+nanomaterials:+what+have+we+learnt+so+far.">Practical considerations for conducting ecotoxicity test methods with manufactured nanomaterials: what have we learnt so far.</a> Ecotoxicology. 21 (4), 933-972 (2012).</li><li>Handy, R. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ecotoxicity+test+methods+for+engineered+nanomaterials:+practical+experiences+and+recommendations+from+the+bench.">Ecotoxicity test methods for engineered nanomaterials: practical experiences and recommendations from the bench.</a> Environmental Toxicology and Chemistry. 31 (1), 15-31 (2012).</li></ol>

浮游植物将太阳能和二氧化碳转化为有机物，因此在水生生态系统中起着关键作用。因此，藻类生长率抑制试验是化学品监管风险评估所需的三项强制性水生毒性试验之一。在这方面，进行可靠和可重复的藻类毒性测试的能力是关键。使用 Erlenmeyer 烧瓶的测试设置引入了一系列变化和不便，如简介 中所述。为了规避这个问题，已经提出了微刺激板3。虽然微刺激板最大限度地减?...

藻类毒性试验是用于生成欧洲和国际法规（例如 REACH 1 和 TSCA（美国））对化学品上市前和上市后注册所需的生态毒性数据所需的三种强制性测试之一。为此，国际组织（如ISO和经合组织）制定了标准化藻类测试准则。这些测试标准和准则在 pH、温度、二氧化碳水平和光强方面规定了理想的测试条件。然而，在藻类试验期间保持稳定的测试条件实际上是困难的，结果存在一系列化学物质和纳米材料（通常称为"困难物质"）的可重复性和可靠性问题2。大多数现有的藻类毒性测试装置都位于孵化器内的轨道摇床上，其体积相对较大（100–250 mL）。这种设置限制了测试浓度和可复制的海藻培养和测试材料的数量。此外，这些设置很少具有均匀的光场，在大型烧瓶中还难以获得可靠的照明条件，部分原因是光强度在光传播得更远时呈指数级下降，部分由于烧瓶几何形状。替代设置包括塑料微刺激器3板，包含小样本量，不允许足够的采样量来测量 pH 值、额外的生物量测量、颜料提取或其他需要破坏性采样的分析。利用现有的纳米材料和形成彩色悬浮物的物质的藻类毒性测试的一个特别挑战是干扰或阻挡藻类细胞可用的光，通常被称为"阴影"4，5。,5测试材料和/或测试材料与藻类细胞之间的相互作用可能发生小瓶内的阴影，或者由于小瓶相对于彼此和光源的定位，在小瓶之间可能发生阴影。
该方法基于阿伦斯伯格等人提出的 小规模藻类毒性测试装置，允许按照经合组织2017和ISO 86928等标准进行测试。该方法得到进一步优化，以解决上述限制：1） 利用 LED 光技术确保均匀的光条件，产生最少的热量;2） 在保持恒定 pH、CO 2 水平的同时，为化学/生物分析提供足够的样品量;3） 允许使用多功能测试容器材料来测试挥发性物质或具有高吸附潜力的物质。

<table>
	<tbody>
		<tr>
			<td>Acetone</td>
			<td>Sigma-Aldrich</td>
			<td>V179124</td>
			<td></td>
		</tr>
		<tr>
			<td>Ammonium chloride</td>
			<td>Sigma-Aldrich</td>
			<td>254134</td>
			<td></td>
		</tr>
		<tr>
			<td>BlueCap bottles (1L)</td>
			<td>Buch &#38; Holm A/S</td>
			<td>&#160;9072335</td>
			<td></td>
		</tr>
		<tr>
			<td>Boric acid</td>
			<td>Sigma-Aldrich</td>
			<td>B0394</td>
			<td></td>
		</tr>
		<tr>
			<td>Calcium chloride dihydrate</td>
			<td>Sigma-Aldrich</td>
			<td>208290</td>
			<td></td>
		</tr>
		<tr>
			<td>Clear acrylic sheet (40x40 cm)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cobalt(II) chloride hexahydrate</td>
			<td>Sigma-Aldrich</td>
			<td>255599</td>
			<td></td>
		</tr>
		<tr>
			<td>Copper(II) chloride dihydrate</td>
			<td>Sigma-Aldrich</td>
			<td>307483</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethylenediaminetetraacetic acid disodium salt dihydrate</td>
			<td>Sigma-Aldrich</td>
			<td>&#160;E5134</td>
			<td></td>
		</tr>
		<tr>
			<td>Fluorescence Spectrophotometer F-7000</td>
			<td>Hitachi</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Hydrochloric acid</td>
			<td>Sigma-Aldrich</td>
			<td>258148</td>
			<td></td>
		</tr>
		<tr>
			<td>Iron(III) chloride hexahydrate</td>
			<td>Sigma-Aldrich</td>
			<td>236489</td>
			<td></td>
		</tr>
		<tr>
			<td>LED light source</td>
			<td>Helmholt Elektronik A/S</td>
			<td>H35161</td>
			<td>Neutral White, 6500K</td>
		</tr>
		<tr>
			<td>Magnesium chloride hexahydrate</td>
			<td>Sigma-Aldrich</td>
			<td>M9272</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnesium sulfate heptahydrate</td>
			<td>Sigma-Aldrich</td>
			<td>230391</td>
			<td></td>
		</tr>
		<tr>
			<td>Manganese(II) chloride tetrahydrate</td>
			<td>Sigma-Aldrich</td>
			<td>221279</td>
			<td></td>
		</tr>
		<tr>
			<td>Orbital shaker</td>
			<td>IKA</td>
			<td>2980200</td>
			<td></td>
		</tr>
		<tr>
			<td>Potassium phosphate monobasic</td>
			<td>Sigma-Aldrich</td>
			<td>P0662</td>
			<td></td>
		</tr>
		<tr>
			<td>Raphidocelis subcapitata</td>
			<td>NORCCA</td>
			<td></td>
			<td>NIVA-CHL1 strain</td>
		</tr>
		<tr>
			<td>Scintillation vials (20 mL)</td>
			<td>Fisherscientific</td>
			<td>11526325</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium bicarbonate</td>
			<td>Sigma-Aldrich</td>
			<td>S6014</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium hydroxide</td>
			<td>Sigma-Aldrich</td>
			<td>415413</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium molybdate dihydrate</td>
			<td>Sigma-Aldrich</td>
			<td>331058&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Spring clamp</td>
			<td>Frederiksen Scientific A/S</td>
			<td>472002</td>
			<td></td>
		</tr>
		<tr>
			<td>Thermostatic cabinet</td>
			<td>VWR</td>
			<td>WTWA208450</td>
			<td>Alternative: temperature controlled room</td>
		</tr>
		<tr>
			<td>Ventilation pipe (&#216;125 mm)</td>
			<td>Silvan</td>
			<td>22605630165</td>
			<td></td>
		</tr>
		<tr>
			<td>Volumetric flasks (25 mL)</td>
			<td>DWK Life Sciences</td>
			<td>246781455</td>
			<td></td>
		</tr>
		<tr>
			<td>Zinc chloride</td>
			<td>Sigma-Aldrich</td>
			<td>208086</td>
			<td></td>
		</tr>
	</tbody>
</table>

a small-scale setup for algal toxicity testing of nanomaterials and other difficult substances

生态毒性数据是欧洲和国际法规（例如 REACH）对化学品进行上市前和上市后注册的要求。藻类毒性试验经常用于化学品的法规风险评估。为了实现高可靠性和可重复性，制定标准化准则至关重要。对于藻类毒性测试，指南要求参数条件稳定且统一，如pH值、温度、二氧化碳水平和光强度。纳米材料和其他所谓的困难物质会干扰光，导致结果变化很大，妨碍了其监管的接受。为了应对这些挑战，我们开发了LEVITATT（藻类毒性试验LED垂直照明表）。该设置利用下面的 LED 照明，实现均匀的光分布和温度控制，同时最大限度地减少样品内阴影。该设置优化了生物量定量的样本量，同时确保 CO2 的充分流入，以支持藻类的指数级增长。此外，可定制测试容器的材料，以最大限度地减少吸附和挥发。在测试彩色物质或颗粒悬浮液时，LED 灯的使用还允许增加光强，而无需额外产生热量。紧凑的设计和最小的设备要求增加了在广泛的实验室中实施 LEVITATT 的可能性。在符合标准化ISO和经合组织藻类毒性测试指南的同时，LEVITATT还显示出两种参考物质（3，5-5-Dicholorophenol和K2Cr2O7）和三种纳米材料（ZnO、CeO2和BaSO4）的样品间变异性低于埃伦迈尔烧瓶和微蒂特板。

对参考物质进行初始测试，以确定藻类菌株的敏感性。经常用于R. 亚卡皮塔的参考物质是二氯酸钾和 3，5-二氯酚7，,8。图 3 和表 2显示了藻类测试的代表性结果，包括曲线拟合和统计输出，当 DRC 包装在 R 中应用于增长率时。
<img alt="Figure 3" class="xfigimg" src="/files/ftp_u...

Watch this Scientific Journal Video about A Small-Scale Setup for Algal Toxicity Testing of Nanomaterials and Other Difficult Substances at JoVE.com

A Small-Scale Setup for Algal Toxicity Testing of Nanomaterials and Other Difficult Substances

我们使用使用 LED 垂直照明的装置，演示对困难物质（如彩色物质或纳米材料）的藻类毒性测试。

纳米材料和其他难物质藻类毒性试验的小型装置

Ecotoxicity data is a requirement for pre- and post-market registration of chemicals by European and international regulations (e.g., REACH). The algal ...

当一家化工公司想要将一种新的化学品或其他材料推向市场时，他们需要提供一系列生态毒性测试的数据。藻类测试是需要执行的三个强制性测试之一。藻类试验受到这样的事实影响，即执行起来相当具有挑战性。因此，我们开发了LEVITATT测试平台。LEVITATT 使用 LED 灯，以避免产生过多的热量问题。此外，它确保了均匀的光场。它具有紧凑的尺寸，这意味着您不需要太多的空间来执行藻类测试，但与此同时，它允许使用足够的样本量在测试之前、期间和测试后执行化学分析和表征。在测试纳米材料时，这一点尤其重要。本视频将演示使用 LEVITATT 对纳米材料和其他困难物质进行藻类毒性测试。演示包括以下四个步骤。首先，描述莱维特的组件。第二步是为藻类生长介质准备解决方案。第三步是概述如何进行藻类毒性试验的过程。最后一步是用荧光计测量藻类生物量的增加。LEVITATT 可放置在温度控制的房间柜中。这允许在整个暴露过程中保持稳定的温度。该设置放置在轨道摇床上，以便测试期间样品旋转。测试在含有4毫升暴露介质的20毫升闪烁小瓶中进行。带钻孔的螺钉盖允许在暴露期间与空气交换 CO2。LED 源夹在两个塑料板之间，位于每个外壳的中心。该位置将创建一个均匀的光场，并最小化样本之间的阴影。每个外壳都装有夹子，最多可以容纳五个样品。紧凑的尺寸可最大限度减少执行实验所需的空间。要准备库存解决方案，请权衡每种化合物的适当量。库存溶液由对藻类生长至关重要的各种盐组成。当适当数量的盐被称重时，它们被超纯水稀释。库存溶液通过过滤或高压灭菌进行灭菌。灭菌后，库存溶液可以储存在4摄氏度的黑暗中。准备一系列暴露浓度。暴露浓度之间的系数不应超过3.2。在每个测试浓度中加入指数级生长藻类的适当体积。最终的细胞密度应该是每毫升大约10，000个藻类细胞。要达到测试的有效性标准，关键是预培养成倍增长。测量小瓶混合，允许藻类和化合物在取样前均匀分布。样品倒入烧杯中，四毫升被移入20毫升的闪烁小瓶中。闪烁小瓶现在被封顶了。从每个浓度移液 0.4 毫升。加入1.6毫升丙酮，扰乱细胞壁，将叶绿素释放至液相。现在每个玻璃瓶都紧闭，以避免在储存过程中丙酮蒸发。封顶闪烁小瓶现在随机分布在LEVITATT中，并孵育72小时。测量孵育开始和结束的光强和温度，以确保符合所需的准则。通过将每个闪烁小瓶中 0.4 毫升的移液到玻璃小瓶中，在 24 小时间隔内进行分采样。加入160万丙酮，破坏细胞壁，将叶绿素释放至液相。添加丙酮后，拧紧盖子，将样品留在黑暗中，直到分析。使用荧光计量化整个测试期间采集的样品中的生物量。荧光仪上的设置应为 420 纳米作为激励波长，发射波长为 671 纳米。每个样本被测量三次，并注意到相对值。荧光计的相对值用于计算将用于进一步数据处理的增长率。使用统计软件将非线性回归曲线拟合到计算的增长率数据中。非线性回归曲线的一个示例可能是可行函数的日志逻辑。关联的 95% 置信区间与模型拟合一起显示。定期有效浓度值（如 10、20 和 50% 低效）用于报告生态毒性测试的结果。总之，LEVITATT为符合国际标准化准则的常规化学品的藻类毒性测试提供了一个紧凑的平台。此外，该装置为测试干扰光向藻类传递的困难物质（如纳米材料或彩色物质）提供了一个强大的平台。

Research

JoVE Journal

Environment

Preparation and Testing of Impedance-based Fluidic Biochips with RTgill-W1 Cells for Rapid Evaluation of Drinking Water Samples for Toxicity

准备和测试阻抗为基础与RTgill-W1细胞饮用水进行快速评估水样进行毒性射流生物芯片

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity ...

科研

环境科学

Experimental Protocol for Biodiesel Production with Isolation of Alkenones as Coproducts from Commercial Isochrysis Algal Biomass

Experimental Protocol for Biodiesel Production with Isolation of Alkenones as Coproducts from Commercial Isochrysis Algal Biomass

实验协议生产生物柴油与饱和酮的分离从商业余积<em&gt;等鞭金藻</em&gt;藻类生物量

The need to replace petroleum fuels with alternatives from renewable and more environmentally sustainable sources is of growing importance. ...

Assaying Predatory Feeding Behaviors in Pristionchus and Other Nematodes

Assaying Predatory Feeding Behaviors in Pristionchus and Other Nematodes

测定掠夺取食行为<em&gt; Pristionchus</em&gt;及其他线虫

This protocol provides multiple methods for the analysis and quantification of predatory feeding behaviors in nematodes. Many nematode species including ...

Extraction of Structural Extracellular Polymeric Substances from Aerobic Granular Sludge

从好氧颗粒污泥结构胞外聚合物的提取

To evaluate and develop methodologies for the extraction of gel-forming extracellular polymeric substances (EPS), EPS from aerobic granular sludge (AGS) ...

Comparison of Scale in a Photosynthetic Reactor System for Algal Remediation of Wastewater

规模在光合反应器系统对污水的整治藻类比较

An experimental methodology is presented to compare the performance of two different sized reactors designed for wastewater treatment. In this study, ...

Functionalization and Dispersion of Carbon Nanomaterials Using an Environmentally Friendly Ultrasonicated Ozonolysis Process

碳纳米材料的功能化和分散使用环境友好的超声波臭氧分解过程

Functionalization of carbon nanomaterials is often a critical step that facilitates their integration into larger material systems and devices. In the ...

Dispersion of Nanomaterials in Aqueous Media: Towards Protocol Optimization

纳米材料在水介质中的分散: 对协议优化的研究

The sonication process is commonly used for de-agglomerating and dispersing nanomaterials in aqueous based media, necessary to improve homogeneity and ...

Harvesting Venom Toxins from Assassin Bugs and Other Heteropteran Insects

从刺客臭虫和其他 Heteropteran 昆虫身上采集毒液毒素

Heteropteran insects such as assassin bugs (Reduviidae) and giant water bugs (Belostomatidae) descended from a common predaceous and venomous ancestor, ...

A High-throughput Assay for the Prediction of Chemical Toxicity by Automated Phenotypic Profiling of Caenorhabditis elegans

A High-throughput Assay for the Prediction of Chemical Toxicity by Automated Phenotypic Profiling of Caenorhabditis elegans

一种用自动表型分析预测线虫化学毒性的高通量测定

Applying toxicity testing of chemicals in higher order organisms, such as mice or rats, is time-consuming and expensive, due to their long lifespan and ...

Development and Testing of Species-specific Quantitative PCR Assays for Environmental DNA Applications

开发和测试针对物种的定量PCR检测环境DNA应用

New, non-invasive methods for detecting and monitoring species presence are being developed to aid in fisheries and wildlife conservation management. The ...