Name: quantification of endogenous auxin and cytokinin during internode culture of ipecac
Uploaded: 2018-03-15T14:00:00
Description: Watch this Scientific Journal Video about Quantification of Endogenous Auxin and Cytokinin During Internode Culture of Ipecac at JoVE.com

我们感谢东洋大学应用生物科学系的 Koudai 先生和群马县农业技术中心谷口先生的技术援助。我们还感谢 Shosaku Kashiwada 教授和东洋大学 Maheswari Rajagopalan 博士的建议。这项研究由东洋大学生命与环境科学研究中心部分支持。

注: 本研究使用催吐剂 (ipecacuanha), 因为它有助于分析内源性激素。
1. 诱导催吐剂不定芽的生长条件
<ol><li>制备无激素的 B5 培养基, 调整为 pH 值为 5.710, 并添加0.2% 结冷胶胶。热处理消毒。</li>
	<li>将25毫升的蒸压介质倒入无菌培养皿 (90 毫米 x 20 毫米)。</li>
	<li>在不育的丙烯酸板上, 用一把22刀片的外科手术刀切开8毫米茎的催吐剂苗, 并放置在培养基上...

<ol>
	<li>Haberlandt, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kulturversuche+mit+isolierten+Pflanzenzellen.">Kulturversuche mit isolierten Pflanzenzellen.</a> Sitzungsber. Math.-Naturwiss. Kl. Akad. Wiss. Wien. 111, 69-92 (1902).</li><li>Skoog, F., Miller, C. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemical+regulation+of+growth+and+organ+formation+in+plant+tissues+cultured+in+vitro.">Chemical regulation of growth and organ formation in plant tissues cultured in vitro.</a> Symp Soc Exp Biol. 11, 118-130 (1957).</li><li>Ganeshan, S., Caswell, K. L., Kartha, K. K., Chibbar, R. N., Khachatourians, G. G., McHughen, A., Scorza, R., Nip, W. 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Richard.</a> Plant Cell Rep. 9 (10), 567-570 (1991).</li><li>Koike, I., Taniguchi, K., Shimomura, K., Umehara, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamics+of+endogenous+indole-3-acetic+acid+and+cytokinins+during+adventitious+shoot+formation+in+ipecac.">Dynamics of endogenous indole-3-acetic acid and cytokinins during adventitious shoot formation in ipecac.</a> J. Plant Growth Regul. , (2017).</li><li>Gamborg, O. L., Miller, R. A., Ojima, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nutrient+requirements+of+suspension+cultures+of+soybean+root+cells.">Nutrient requirements of suspension cultures of soybean root cells.</a> Exp. Cell Res. 50 (1), 151-158 (1968).</li><li>Watad, A. 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D. <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+of+cytokinins+on+in+vitro+propagation+of+Bacopa+monniera.">Comparative studies of cytokinins on in vitro propagation of Bacopa monniera.</a> Plant Cell Tiss. Org. Cult. 66 (1), 9-16 (2001).</li><li>Rao, M. S., Purohit, S. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+shoot+bud+differentiation+and+plantlet+regeneration+in+Celastrus+paniculatus+Willd.">In vitro shoot bud differentiation and plantlet regeneration in Celastrus paniculatus Willd.</a> Biol. Plant. 50 (4), 501-506 (2006).</li><li>Sanikhani, M., Frello, S., Serek, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=TDZ+induces+shoot+regeneration+in+various+Kalancho&#235;+blossfeldiana+Poelln+cultivars+in+the+absence+of+auxin.">TDZ induces shoot regeneration in various Kalancho&#235; blossfeldiana Poelln cultivars in the absence of auxin.</a> Plant Cell Tiss. Org. Cult. 85 (1), 75-82 (2006).</li><li>Yoshimoto, K., 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=Autophagy+negatively+regulates+cell+death+by+controlling+NPR1-dependent+salicylic+acid+signaling+during+senescence+and+the+innate+immune+response+in+Arabidopsis.">Autophagy negatively regulates cell death by controlling NPR1-dependent salicylic acid signaling during senescence and the innate immune response in Arabidopsis.</a> Plant Cell. 21 (9), 2914-2927 (2009).</li><li>Schaller, G. E., Bishopp, A., Kieber, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+yin-yang+of+hormones:+cytokinin+and+auxin+interactions+in+plant+development.">The yin-yang of hormones: cytokinin and auxin interactions in plant development.</a> Plant Cell. 27 (1), 44-63 (2015).</li></ol>

为了确定参与器官激素的分布, 重要的是使用植物材料在无激素培养基上观察器官, 因为当激素被外部应用于外植体诱导芽或根, 它们影响整个外植体, 使得很难评价植物的内在可塑性的细胞分化和器官。不定芽可诱导其他植物的无激素培养基 (如石竹石竹L.11、 Aegle marmelos (l.) Corrêa12、 Bacopa 床子(l.) Pennell13, 藤苋菜。<s...

Haberlandt (1854-1945) 提出了 "全能" 的概念, 在成熟植物中, 植物细胞可以分化、分化和再生整个植物, 甚至在它们的前向特定细胞类型的差异1。在组织培养中, 能否诱导植物再生是由外部在生长培养基中应用激素的组合和浓度决定的。Skoog 和米勒发现, 在含有高中正比植物生长素的培养基上, 烟草愈伤组织可诱导不定芽, 而不定根可在含有低比值2 的培养基上诱导。自此发现以来, 组织培养已广泛用于经济重要作物的繁殖和转基因植物的再生3。不定芽可诱导从其他组织以外的茎尖分生, 如叶, 根, 和节间。在大多数植物中诱导不定芽需要激素治疗。然而, 最佳的浓度和组合不同的物种和组织之间作为外植体。因此, 大量的努力去确定激素的最佳浓度和组合的实验。
Carapichea ipecacuanha(Brot)l. 安德森 (催吐剂) 是一种药用植物, 含有吐根碱和 cephaeline 等生物碱, 主要在根4 中。根提取物用作祛痰、催吐剂和 amoebicide5。虽然催吐剂自然生长在巴西的热带雨林中, 但它不愿意在文化中设置种子, 在日本的种子储藏过程中, 发芽率下降, 气候较冷的6。相反, 它传播的组织培养, 在节间不定的拍摄形成是最有效的方法7,8。有趣的是, 不定芽可以诱导在这个物种没有激素治疗8。
不定芽形成于茎节顶端区域的表皮上, 不愈伤组织, 但不在基底区域9。这种差异表明茎段的组织极性, 这可能是在 phytohormonal 调节。催吐剂培养系统允许一个独特的机会来分析在不定芽形成期间内源激素水平的变化。在这里, 我们介绍了分析一个生长素 (indole-3-acetic 酸 (IAA)) 和四中正 (isopentenyl 腺嘌呤 (iP), isopentenyl 腺嘌呤核苷 (iPR),反式-玉米 () 和反式-玉米核苷 (tZR) 的内生水平的方法。在茎段中通过使用 LC/ms。

<table border="0" cellpadding="0" cellspacing="0" style="width:677px;" width="678">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">[2H5]indole-3-acetic acid</td>
			<td style="width:176px;">Olchemlm Ltd</td>
			<td style="width:119px;">031 1531</td>
			<td style="width:167px;">Internal standard for LC-MS/MS</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">[2H5]trans-zeatin</td>
			<td style="width:176px;">Olchemlm Ltd</td>
			<td style="width:119px;">030 0301</td>
			<td>Internal standard for LC-MS/MS</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">[2H5]trans-zeatin riboside</td>
			<td style="width:176px;">Olchemlm Ltd</td>
			<td style="width:119px;">030 0311</td>
			<td>Internal standard for LC-MS/MS</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">[2H6]N6-isopentenyl adenine</td>
			<td style="width:176px;">Olchemlm Ltd</td>
			<td style="width:119px;">030 0161</td>
			<td>Internal standard for LC-MS/MS</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">[2H6]N6-isopentenyl adenosine</td>
			<td style="width:176px;">Olchemlm Ltd</td>
			<td style="width:119px;">030 0171</td>
			<td>Internal standard for LC-MS/MS</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">indole-3-acetic acid</td>
			<td style="width:176px;">Wako</td>
			<td>098 00181</td>
			<td>standard for LC-MS/MS</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">trans-zeatin</td>
			<td style="width:176px;">SIGMA-ALDRICH</td>
			<td style="width:119px;">Z0876 5MG</td>
			<td>standard for LC-MS/MS</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">trans-zeatin riboside</td>
			<td style="width:176px;">Wako</td>
			<td>262 01081</td>
			<td>standard for LC-MS/MS</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">N6-isopentenyl adenine</td>
			<td style="width:176px;">SIGMA-ALDRICH</td>
			<td style="width:119px;">D7674 1G</td>
			<td>standard for LC-MS/MS</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">N6-isopentenyl adenosine</td>
			<td style="width:176px;">ACROS ORGANICS</td>
			<td style="width:119px;">22648 1000</td>
			<td>standard for LC-MS/MS</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">acetonitrile hypergrade for LC-MS LiChrosolv</td>
			<td style="width:176px;">MERCK</td>
			<td style="width:119px;">1.00029.1000</td>
			<td>solvent for LC-MS/MS</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Water for chromatography LiChrosolv</td>
			<td style="width:176px;">MERCK</td>
			<td style="width:119px;">1.15333.1000</td>
			<td>solvent for LC-MS/MS</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">HPLC</td>
			<td style="width:176px;">SHIMADZU</td>
			<td style="width:119px;">Prominence</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">MS</td>
			<td style="width:176px;">Sciex</td>
			<td style="width:119px;">3200QTRAP</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Oasis HLB 30 mg/1 cc</td>
			<td style="width:176px;">Waters</td>
			<td style="width:119px;">WAT094225</td>
			<td>cartridge column</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Oasis MCX 30 mg/1 cc</td>
			<td style="width:176px;">Waters</td>
			<td style="width:119px;">186000252</td>
			<td>cartridge column</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">screw neck total recovery vial</td>
			<td style="width:176px;">Waters</td>
			<td style="width:119px;">186002805</td>
			<td></td>
		</tr>
		<tr height="61">
			<td height="61" style="height:61px;width:216px;">blue, 12 x 32mm screw neck cap and PTFE/silicone septum</td>
			<td style="width:176px;">Waters</td>
			<td style="width:119px;">186000274</td>
			<td></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Acquity UPLC BEH C18, 2.1x100 mm</td>
			<td style="width:176px;">Waters</td>
			<td style="width:119px;">186002350</td>
			<td>UPLC column</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Proshell 120 EC-C18, 2.1x50 mm</td>
			<td style="width:176px;">Agilent</td>
			<td style="width:119px;">699775-902</td>
			<td>UPLC column</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Digital microscope</td>
			<td style="width:176px;">Leica</td>
			<td style="width:119px;">DHS1000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">TissueLyser II</td>
			<td style="width:176px;">QIAGEN</td>
			<td style="width:119px;">85300</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Surgical blade</td>
			<td style="width:176px;">Feather</td>
			<td style="width:119px;">No. 22</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Scalpel handle</td>
			<td style="width:176px;">Feather</td>
			<td style="width:119px;">No. 4</td>
			<td></td>
		</tr>
		<tr height="61">
			<td height="61" style="height:61px;width:216px;">Savant SpeedVac/Refregerated vapor trap</td>
			<td style="width:176px;">Thermo Fisher Scientific</td>
			<td style="width:119px;">SPD111/RVT4104</td>
			<td>vacuum concentrartor</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Disposable glass tobe (13x100 mm)</td>
			<td style="width:176px;">IWAKI</td>
			<td style="width:119px;">9832-1310</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Sterile petri dish</td>
			<td style="width:176px;">INA OPTICA</td>
			<td style="width:119px;">I-90-20</td>
			<td></td>
		</tr>
	</tbody>
</table>

quantification of endogenous auxin and cytokinin during internode culture of ipecac

不定芽形成是重要的技术, 对重要作物的繁殖和转基因植物的再生。在大多数物种中诱导不定芽需要激素治疗。不定芽能否诱发是由生长素与细胞分裂素 (CK) 水平的平衡决定的。大量的努力, 以确定最佳的浓度和组合激素在每个组织作为外植体和在每个植物物种。在催吐剂中, 无激素处理可诱导培养基中茎段的不定芽。这使得催吐剂的内在可塑性可以被评估。为了诱导催吐剂中的不定芽, 我们在15µmol m−2 s−1的茎段中培养了24摄氏度, 在14小时 light/10-h 黑循环中, 在无激素 B5 培养基上固化0.2% 结冷胶胶5周。通过液相色谱-串联质谱 (LC ms) 测定了不定芽形成过程中的激素动态, 并对 indole-3-acetic 中的内源性酸和中正进行了检测。这种方法可以简单地分析内源性 indole-3-acetic 酸和中正水平。可用于研究其他植物器官中内源生长素和 CK 的动态变化。

在 1st周, 没有形成不定芽。在 2nd周, 出现了小芽。在 3rd和 4th周, 芽数增加主要在顶端区域 (I 和 II) (图 2A)。在 5th周, 芽的数量在区域 i 和 5 (图 2B) 中大约为7。相比之下, 第三和 iv. 区只形成了几枝芽。
在培养之前, IA...

Watch this Scientific Journal Video about Quantification of Endogenous Auxin and Cytokinin During Internode Culture of Ipecac at JoVE.com

Quantification of Endogenous Auxin and Cytokinin During Internode Culture of Ipecac

催吐剂节间培养中内源生长素和细胞分裂细胞的量化

无激素治疗的催吐剂茎段可诱发不定芽。为了评价不定芽形成过程中的激素动态, 我们用 LC-ms/ms 测定了茎段内源性生长素和细胞分裂素。

Adventitious shoot formation is an important technique for the propagation of economically important crops and for the regeneration of transgenic plants. ...

The overall goal of this procedure is to describe how endogenous phytohormone levels change during adventitious shoot formation in ipecac. This method can help answer key questions related to tissue regeneration in the plant tissue culture wheel. The main advantage of this technique is the endogenous level of Indole-3-acetic acid and cytokinines can be simply analyzed with high sensitivity.The implication of this technique extend throughout establishment of plant tissue degeneration because changes in endogenous phytohormone level trigger adventitious shoot formation. To begin, prepare a phytohormone-free B5 medium and add 0.2 percent gellan gum. Then, autoclave the medium to sterilize it.Using a surgical scalpel on a sterile acrylic plate, slice eight millimeter internodal segments of ipecac plantlets and place them on the medium. Then, culture the ipecac segments at 24 degrees Celsius under 15 micromole per square meter seconds of light in a 14-hour light, 10-hour dark cycle for five weeks. After culturing the ipecac segments, identify regions one through four in each segment.Once a week, use a microscope to count the number of adventitious shoots in each region. First, place a five millimeter zirconia bead into four two-milliliter sample tubes. Then, use a surgical scalpel on a sterile acrylic plate to cut the ipecac segments into regions I through IV.Then collect eight segments from each region into one sample tube.Weigh each of the samples and freeze them in liquid nitrogen. Next, use a bead-based homogenizer to crush the frozen samples. Use a vortex mixer to suspend the samples in one milliliter of the acetonitrile with 500 picograms of each internal standard of oxen and CKs.Home the samples at four degrees Celsius for one hour. And then centrifuge the samples at 3500 G for five minutes at room temperature. Wash the pellet in 80 percent acetal nitrile with one percent acetic acid.Then repeat the centrifugation and use a disposable class tube to combine the supernatants. Add 600 microliters of water with one percent acetic acid to the combined supernatant. Then use a vacuum concentrator to evaporate the acetal nitrile.Load the sample solutions into individual equilibrated HLB cartridges. And wash the cartridges with one milliliter of water with one percent acetic acid. Then allude all of the hormones with two milliliters of 80 percent acetyl nitrile with one percent acetic acid in a glass tube.Using a vacuum concentrator, evaporate the acetal nitrile to obtain extract in water with one percent acetic acid. Load the sample solutions into individual equilibrated MCX cartridges and wash the cartridges with one milliliter of water with one percent acetic acid. Then allude IAA in a glass tube, with two milliliters of 30 percent acetal nitrile with one percent acetic acid.Then wash the cartridges with two milliliters of 80 percent acetal nitrile with one percent acetic acid. Next, wash the cartridges with two milliliters of water and one milliliter of water with five percent aqueous ammonia in a draft chamber. Allude the CKs in a glass tube with two milliliters of 60 percent acetal nitrile with five percent aqueous ammonia in a draft chamber.Then use a vacuum concentrator to evaporate the solvent of each hormone fraction. Stir the samples at minus thirty degrees Celsius until LCMSMS analysis. First, dissolve each hormone extract in a tube with 600 microliters of methanol.Then transfer the solution to a screw-neck total recovery vial and use a vacuum concentrator to evaporate the solvent. Dissolve the IAA fraction in 20 microliters of 30 percent acetal nitrile and the CK fractions in 20 microliters of water with one percent acetic acid. Next use a triple quadrupole MS system equipped with an HPLC system to analyze the samples in positive ion mode.Finally quantify endogenous IAA in CK levels against a standard curve of the ration of unlabeled to deuterium label standards. In this protocol endogenous oxidant cytokinin in adventitious chutes was measured via LCMSMS for five weeks. At the first week, no adventitious chutes had formed and the first small chutes appeared during the second week.During the third and fourth weeks, the number of chutes increased mostly in regents one and two. During the fifth week the number of chutes was approximately seven in region one, and five in region two. Before culture, the IAA level was slightly higher in region one than in regions two, three, and four.During the first week, the IAA level increased greatly in region four, and decreased slightly in regions one, two, and three. By five weeks of culture an IAA concentration gradient emerged. With levels increasing from region one to region four.Before culture, there were only trace levels of most of the CKs. At the first week, the tZR level increased to 13.8 picograms per milligram in region two. And 18.1 picograms per milligram in region three.In contrast the levels of tZ, iP and iPR changed only slightly during culture. While attempting this procedure it's important to remember that endogenous part hormone are stable. After it's developed man, this technique paves the way for researchers in the field of plant tissue culture.To explore adventitious formation in other organisms. After watching this video, you should have a good understanding of how to analyze endogenous Indole-3-acetic acid and cytokinines.

Research

JoVE Journal

Environment

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. I. Collection of Virus Samples

EPA方法1615测量肠道病毒和诺瓦克病毒发生水由文化和RT-qPCR的了。病毒样本的收集I.

EPA Method 1615 was developed with a goal of providing a standard method for measuring enteroviruses and noroviruses in environmental and drinking waters. ...

科研

环境科学

Quantification of Heavy Metals and Other Inorganic Contaminants on the Productivity of Microalgae

重金属和微藻的产其他无机污染物的量化

Increasing demand for renewable fuels has researchers investigating the feasibility of alternative feedstocks, such as microalgae. Inherent advantages ...

Simultaneous DNA-RNA Extraction from Coastal Sediments and Quantification of 16S rRNA Genes and Transcripts by Real-time PCR

从海岸沉积物和16S量化的rRNA基因和转录同时DNA-RNA提取采用Real-time PCR检测

Real Time Polymerase Chain Reaction also known as quantitative PCR (q-PCR) is a widely used tool in microbial ecology to quantify gene abundances of ...

Capturing Flow-weighted Water and Suspended Particulates from Agricultural Canals During Drainage Events

在排水过程中捕获农用运河中的流动加权水和悬浮颗粒

The purpose of this study is to describe the methods used to capture flow-weighted water and suspended particulates from farm canals during drainage ...

The Effect of the Application of Thyme Essential Oil on Microbial Load During Meat Drying

百里香精油应用于肉类干燥过程中微生物负荷的影响

Meat is a high protein meal that is used in the preparation of jerky, a popular food snack, where preservation and safety are important. To assure food ...

Stress Distribution During Cold Compression of Rocks and Mineral Aggregates Using Synchrotron-based X-Ray Diffraction

基于同步辐射 x 射线衍射的岩石和矿物团聚体冷压缩应力分布

We report detailed procedures for performing compression experiments on rocks and mineral aggregates within a multi-anvil deformation apparatus (D-DIA) ...

Chemical Isolation, Quantification, and Separation of Skin Lipids from Reptiles

爬行动物皮肤脂类的化学分离、定量和分离

Reptiles signal to conspecifics using lipids in their skin, primarily to enable mate tracking and assessment. The isolation of these lipids has utility in ...

Split Point Analysis and Uncertainty Quantification of Thermal-Optical Organic/Elemental Carbon Measurements

热光学有机/元素碳测量的分割点分析与不确定性定量

Researchers from myriad fields frequently seek to quantify and classify concentrations of carbonaceous aerosols as organic carbon (OC) or elemental carbon ...

Processing Embryo, Eggshell, and Fungal Culture for Scanning Electron Microscopy

用于扫描电子显微镜的胚胎、蛋壳和真菌培养

Although scanning electron microscopy (SEM) is being widely used for the ultra-structural analysis of various biological and non-biological samples, ...