这项工作是由来自教育，青年，体育和（LO1304）捷克外交部和捷克共和国的技术局（TE01020028）资助。笔者想感谢拉克什曼瓦拉纳西博士采取环保盖的静止图像。

1.琼脂糖包被的平板的制备<ol><li>权衡0.75克低熔点琼脂糖和它的McCoy的5A培养基中的溶液（有或没有酚红）无血清添加到100。加热在微波的溶液，摇动每1-2分钟以完全溶解琼脂糖。高压灭菌的溶液消毒它。 </li><li>冷却该高压灭菌琼脂糖至约70℃，并通过500毫升，0.22微米过滤器顶部通过真空过滤在层流箱过滤。如果不使用整个解决方案立刻分装0.75％的过滤琼脂糖溶液（FAS）为更小的体积。在寒冷的房间或4℃冰箱中长达4周存储这个准备使用的琼脂糖溶液无菌。 </li><li>附加一个小管与胶面纸或金属嘴分配盒到的Combi试剂分配器和素盒与一个流浆箱70％的乙醇（乙醇），然后用无菌磷酸盐缓冲盐水（PBS）。 </li><li>在使用前，加热FAS的i存储的等分试样<html> n个微波融化它。总理与琼脂糖溶液和外套384孔，组织培养（TC）分配盒 - 处理"特殊"与微孔板FAS的15微升。允许在板琼脂糖接种细胞之前冷却15-20分钟。存储琼脂糖涂层板在无菌条件下，包裹在聚乙烯袋中在寒冷的房间或在4℃，并远离直射光。 注:避免0.75％的FAS的反复加热，以防止在琼脂糖中的储备溶液浓度的变化。当在上述条件下保存的琼脂糖包被的板可以存储最多2周。上FAS板的涂覆过程中不需要加热。 </li><li>通过用70-80℃无菌水吸它以除去任何剩余的琼脂糖在盒提示和管清洁分配盒。 </li></ol> 2.细胞培养和形成MCTS <ol><li>培养人结肠直肠癌HCT116细胞，如先前所描述姑娘="外部参照"&gt; 8。 </li><li>除去所需数量的琼脂糖包被，384孔，从冷库TC-处理微孔板和它们平衡至室温（RT）搅拌15分钟。 </li><li>通过吸用70％乙醇和无菌PBS中的标准管分配盒制备用于细胞接种的组合式试剂分配器。调节播种卷到所需微升和使用上的分配器手动设定按钮分配速度到介质。 注:在无菌条件下，并在层流箱中进行整个细胞接种过程。 </li><li>分离使用重组细胞解离酶从组织培养瓶中的贴壁细胞。使在与种子细胞的无菌烧杯的细胞悬浮液的库存在完全生长培养基的50微升一个的每孔2.5×10 4个细胞/ mL的密度。当播种多个384可湿性粉剂，使用磁力搅拌器以防止它们沉降到烧杯的底部搅动细胞。 </li><li>允许板休息在RT 30分钟，然后离心以4×g下15分钟。 </li><li>同时，采取蒸发减少环境盖和用8毫升无菌H型2 O或5％二甲亚砜（DMSO）中的短边填充它使用5毫升吸管（ 图1A）（左和右）。首先，分配通过扫描枪头慢慢上下充填液体进入左侧槽的4毫升。重复此步骤以右侧槽。 注:确保液体加入到侧槽的盖子的中心不会合并，并留下气体交换（ 图1A）的差距。加入的H成过量2 O的结果在H 2 O的渗流到盖的外部，并随后到384-孔TC板的外孔。 </li><li>填用无菌H 2澳384孔TC板中的液体贮存器和替换用充满液体的环境盖子的常规板盖（ 图2A）。 </li><li>放置在用95％的湿度，5％CO 2和20％O 2 37℃的旋转恒温箱板，并且允许细胞聚集成MCTSes 4天。避免为了防止突然跌落湿度水平打开培养箱门太久在随后的日子里。 </li></ol> 3.中使用外汇机器人系统<ol><li>在下面的MCTS形成的第4天，每口井使用自动洗板机饮水机加预热中的30微升。允许MCTSes中生长额外3天。更换培养基每3天定期，直到它们达到所需的大小进行实验。 </li><li>为了吸定义从每个介质的体积以及凭经验调整垫圈歧管的Z -height。吸30％以及介质μL，并与新鲜，预热介质的30微升更换。 注意:在该垫圈歧管向下传播设置配送速度和速度成以最低速率孔，以尽量减少在孔中的湍流。 </li></ol> MCTS 4.高内涵成像和半自动图像分析<ol><li>图像的MCTS在使用4X空中目标的内容高度自动化成像系统（NA 16）。 </li><li>设置曝光时间11毫秒和分级，以4×4，调节的Z -stacks的数目和间隔并根据需要进行实验以每孔捕获整个MCTS的像素组合。 </li><li>处理图像逐步，如在所描述的"自述"，使用写在编程语言中的内部程序。 注:.M代码和.txt自述文件可作为补充代码文件（ 图1B）。常规措施MCTS面积，长轴和短轴，周长和坚固由2D图像。 </li></ol><img alt="图1" src="/files/ftp_upload/55403/55403fig1.jpg" /> 图1:环境罐盖的制备和半自动程序的流程图。 （A）中填充有正确的（左）和过量（右）填充液体的量（这里，5％DMSO）中的蒸发减少对环境的盖的图像。箭头指示用于将液体中的短边槽。星号示出了加入DMSO的正确体积的以下在盖中间的间隙。 （B）中示出所涉及的半自动化图像分析程序中的步骤的工作流程。 <a href="http://ecsource.jove.com/files/ftp_upload/55403/55403fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

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作者什么都没有透露。

涂层384孔TC板用过滤琼脂糖在很多的标准做法是使用1-1.5％低熔点琼脂糖涂覆板，这需要琼脂糖和/或分配单元也可以保持加热，以防止琼脂糖6的胶凝。琼脂糖的凝胶化是可能引起关注的，而使用具有小管的孔的范围直径在0.2和0.4之间的液体分配盒制备多块板。为了克服堵塞分配盒的潜在问题，0.75％的FAS被使用，因为它在分配期间不需要额外的加热。顺便说一?...

在肿瘤癌细胞生理学布置在一个复杂的，三维（3D）细胞外基质和相互作用的细胞所包围的结构。作为近组织中的所有细胞驻留在3D环境中，有必要在该模拟肿瘤性状体外肿瘤模型更生理学相关导致的几个三维培养技术1，2，3的发展。这些模型正在成为基本的研究工具用于研究在三维2转移和细胞响应于治疗肿瘤微环境的作用。此外，相对于2维（2D）细胞培养物4，3D模型允许更好地了解肿瘤-基质的相互作用，从而影响细胞信号通路的。 的肿瘤细胞株多细胞肿瘤球状体（MCTSes）在三维细胞c的常用由于其相对接近体内肿瘤ulture车型。出的使用中的多种技术，MCTS代的琼脂糖包被的平板的液体覆盖技术（LOT）已经获得了铅的优化和靶标确认5，6，7，8，9，10，11显著兴趣。这是从最近的研究已成功能在使用LOT 6,7 MCTS文化运行化合物文库的导频屏幕明显。然而，由于介质的蒸发引起的不均匀损耗油井到井变异MCTS形态学和生长是伴随使用微量滴定板（MPS）的LOT常见障碍。因此，非均匀MCTSes形成危及意义和相关从药理学试验8，12，13的数据。除了再现性的问题，使用自动液体分配单元时影响基于LOT-高通量测定另一实际问题是在MP的涂布用琼脂糖。虽然分配单元可以保持加热，以防止琼脂糖的凝胶化，分配盒和管道的堵塞是机器人系统6的电位的关注。 为了克服这些挑战，我们最近在很多关于MCTS文化8设计了一些修改。这些修改主要是根据可能的方法来防止用户使用该工具在高通量筛查实验室中常见的国会议员中损失不均匀。修改后的很多跨3大小均匀，重现性MCTSes生成的详细过程84孔板（WPS）这里提出。手稿还提出了MCTS大小的评价半自动程序，特别是在不具有横截面面积的测量一个明确定义的边界部分破碎，药物处理MCTSes。

<table><tbody><tr><td>Agarose</td><td>Sigma-Aldrich</td><td>A9414</td><td>Low-melting</td></tr><tr><td>McCOY&#039;s 5A Medium</td><td>Sigma-Aldrich</td><td>M8403</td><td></td></tr><tr><td>&amp;ldquo;rapid&amp;rdquo; Filtermax filter</td><td>TPP</td><td>99505</td><td>0.22 &amp;mu;m, 500 mL</td></tr><tr><td>Multidrop&amp;trade; Combi Reagent Dispenser&amp;nbsp;</td><td>Thermo Fisher Scientific</td><td>5840300</td><td></td></tr><tr><td>Small Tube Dispensing cassette&amp;nbsp;</td><td>Thermo Fisher Scientific</td><td>24073295</td><td>Metal tip&amp;nbsp;</td></tr><tr><td>384-well TC plate&amp;nbsp;</td><td>PerkinElmer</td><td>6057308</td><td>Plate type- CellCarrier</td></tr><tr><td>Standard Tube Dispensing Cassette</td><td>Thermo Fisher Scientific</td><td>24072670</td><td></td></tr><tr><td>MicroClime Environmental Lid</td><td>Labcyte</td><td>LLS-0310</td><td></td></tr><tr><td>DMSO</td><td>Sigma</td><td>D4540</td><td></td></tr><tr><td>Rotary Incubator (SteriStore )</td><td>HighRes Biosolutions</td><td>23641</td><td>Serial No.: D00384</td></tr><tr><td>Microplate Washer Dispenser&amp;nbsp;</td><td>BioTek</td><td>Unspecified</td><td>Model: EL406&amp;nbsp;</td></tr><tr><td>High-Content Imaging System (CellVoyager )</td><td>Yokogawa Electric Corporation</td><td>Unspecified</td><td>Model: CV7000</td></tr><tr><td>Orange G</td><td>New England Biolabs</td><td>B7022S</td><td></td></tr><tr><td>TrypLE&amp;trade; Express recombinant cell dissociation reagent</td><td>Thermo Fisher Scientific</td><td>12604021</td><td>Phenol red free</td></tr></tbody></table>

evaporation-reducing culture condition increases the reproducibility of multicellular spheroid formation in microtiter plates

肿瘤模型， 在体内条件密切模仿正成为药物发现和开发的潜在的抗癌药物的筛选日益流行。多细胞肿瘤球状体（MCTSes）有效模仿实体瘤的生理条件下，使它们在体外模型优良铅优化和目标验证。从可供MCTS文化的各种技术，在琼脂糖液体覆盖法是对MCTS一代最便宜的方法之一。然而，使用液体的覆盖为高通量筛选MCTS培养物的可靠传输可能由许多限制，包括微量滴定板（MPS）琼脂糖的涂层并在井均匀MCTS地层的不可再现性受到损害。国会议员是显著容易发生边缘从培养基从板的外部过度蒸发造成的影响，防止药物使用整个板块的试验。这个手稿提供到液体覆盖技术来增加均匀MCTS形成的可扩展性和可重复性的详细技术的改进。此外，药物治疗提出后，对MCTS的评价简​​单，半自动，和普遍适用的软件工具的细节特征。

不均匀的损失中，尤其是来自周边井，是小培养体积议员们经常遇到的问题。大大改善培养条件，如与良好控制的温度/加湿系统和蒸发减少国会议员和板盖孵化器，降低介质的整个井8的显著损失。测量相对蒸发，橙G（OG）等体积加入到每孔中，并用3天OG吸光度变化被记录在与在标准和旋转孵化定期和环境盖板。板孔被分成基于从板（ 图2A）...

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Evaporation-reducing Culture Condition Increases the Reproducibility of Multicellular Spheroid Formation in Microtiter Plates

培养基从微量滴定板的凹凸损失影响均匀的多细胞肿瘤球体形成的再现性。改善培养条件，以减少显著介质损耗将提高球体形成的再现和基于球状体的测定的使用液体覆盖技术的结果。

蒸发减少培养条件的提高多细胞球体形成的微孔板重现

Tumor models that closely imitate in vivo conditions are becoming increasingly popular in drug discovery and development for the screening of potential ...

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JoVE Journal

Cancer Research

6.9K 次观看.  Palacky University in Olomouc. 培养基从微量滴定板的凹凸损失影响均匀的多细胞肿瘤球体形成的再现性。改善培养条件，以减少显著介质损耗将提高球体形成的再现和基于球状体的测定的使用液体覆盖技术的结果。 

视频: Evaporation-reducing Culture Condition Increases the Reproducibility of Multicellular Spheroid Formation in Microtiter Plates

Production of Large Numbers of Size-controlled Tumor Spheroids Using Microwell Plates

Tumor spheroids are increasingly recognized as an important in vitro model for the behavior of tumor cells in three dimensions. More physiologically relevant than conventional adherent-sheet cultures, they more accurately recapitulate the complexity and interactions present in real tumors. In order to harness this model to better assess tumor biology, or the efficacy of novel therapeutic agents, it is necessary to be able to generate spheroids reproducibly, in a controlled manner and in significant numbers.
The AggreWell system consists of a high-density array of pyramid-shaped microwells, into which a suspension of single cells is centrifuged. The numbers of cells clustering at the bottom of each microwell, and the number and ratio of distinct cell types involved depend only on the properties of the suspension introduced by the experimenter. Thus, we are able to generate tumor spheroids of arbitrary size and composition without needing to modify the underlying platform technology. The hundreds of microwells per square centimeter of culture surface area in turn ensure that extremely high production levels may be attained via a straightforward, nonlabor-intensive process. We therefore expect that this protocol will be broadly useful to researchers in the tumor spheroid field.

We introduce a protocol for the generation of large numbers (thousands to hundreds of thousands) of uniform size- and composition-controlled tumor spheroids, using commercially available microwell plates.

生产尺寸控制肿瘤细胞球体采用微孔板大数

Tumor spheroids are increasingly recognized as an important in vitro model for the behavior of tumor cells in three dimensions. More physiologically ...

科研

生物工程

Assessing Cell Viability and Death in 3D Spheroid Cultures of Cancer Cells

Three-dimensional spheroids of cancer cells are important tools for both cancer drug screens and for gaining mechanistic insight into cancer cell biology. The power of this preparation lies in its ability to mimic many aspects of the in vivo conditions of tumors while being fast, cheap, and versatile enough to allow relatively high-throughput screening. The spheroid culture conditions can recapitulate the physico-chemical gradients in a tumor, including the increasing extracellular acidity, increased lactate, and decreasing glucose and oxygen availability, from the spheroid periphery to its core. Also, the mechanical properties and cell-cell interactions of in vivo tumors are in part mimicked by this model. The specific properties and consequently the optimal growth conditions, of 3D spheroids, differ widely between different types of cancer cells. Furthermore, the assessment of cell viability and death in 3D spheroids requires methods that differ in part from those employed for 2D cultures. Here we describe several protocols for preparing 3D spheroids of cancer cells, and for using such cultures to assess cell viability and death in the context of evaluating the efficacy of anticancer drugs.

Here, we present several simple methods for evaluating viability and death in 3D cancer cell spheroids, which mimic the physico-chemical gradients of in vivo tumors much better than the 2D culture. The spheroid model, therefore, allows evaluation of the cancer drug efficacy with improved translation to in vivo conditions.

评估癌细胞3D球形培养中的细胞活力和死亡

Three-dimensional spheroids of cancer cells are important tools for both cancer drug screens and for gaining mechanistic insight into cancer cell biology. ...

癌症研究

Exploring Mitochondrial Energy Metabolism of Single 3D Microtissue Spheroids Using Extracellular Flux Analysis

Three-dimensional (3D) cellular aggregates, termed spheroids, have become the forefront of in vitro cell culture in recent years. In contrast to culturing cells as two-dimensional, single-cell monolayers (2D culture), spheroid cell culture promotes, regulates, and supports physiological cellular architecture and characteristics that exist in vivo, including the expression of extracellular matrix proteins, cell signaling, gene expression, protein production, differentiation, and proliferation. The importance of 3D culture has been recognized in many research fields, including oncology, diabetes, stem cell biology, and tissue engineering. Over the last decade, improved methods have been developed to produce spheroids and assess their metabolic function and fate.
Extracellular flux (XF) analyzers have been used to explore mitochondrial function in 3D microtissues such as spheroids using either an XF24 islet capture plate or an XFe96 spheroid microplate. However, distinct protocols and the optimization of probing mitochondrial energy metabolism in spheroids using XF technology have not been described in detail. This paper provides detailed protocols for probing mitochondrial energy metabolism in single 3D spheroids using spheroid microplates with the XFe96 XF analyzer. Using different cancer cell lines, XF technology is demonstrated to be capable of distinguishing between cellular respiration in 3D spheroids of not only different sizes but also different volumes, cell numbers,&#160;DNA content and type.
The optimal mitochondrial effector compound concentrations of oligomycin, BAM15, rotenone, and antimycin A are used to probe specific parameters of mitochondrial energy metabolism in 3D spheroids. This paper also discusses methods to normalize data obtained from spheroids and addresses many considerations that should be considered when exploring spheroid metabolism using XF technology. This protocol will help drive research in advanced in vitro spheroid models.

These protocols will help users probe mitochondrial energy metabolism in 3D cancer cell-line-derived spheroids using Seahorse extracellular flux analysis.

利用细胞外通量分析探索单个3D微组织球体的线粒体能量代谢

Three-dimensional (3D) cellular aggregates, termed spheroids, have become the forefront of in vitro cell culture in recent years. In contrast to culturing ...

Establishing a High Throughput Epidermal Spheroid Culture System to Model Keratinocyte Stem Cell Plasticity

Epithelial dysregulation is a node for a variety of human conditions and ailments, including chronic wounding, inflammation, and over 80% of all human cancers. As a lining tissue, the skin epithelium is often subject to injury and has evolutionarily adapted by acquiring the cellular plasticity necessary to repair damaged tissue. Over the years, several efforts have been made to study epithelial plasticity using in&#160;vitro and ex vivo cell-based models. However, these efforts have been limited in their capacity to recapitulate the various phases of epithelial cell plasticity. We describe here a protocol for generating 3D epidermal spheroids and epidermal spheroid-derived cells from primary neonatal human keratinocytes. This protocol outlines the capacity of epidermal spheroid cultures to functionally model distinct stages of keratinocyte generative plasticity and demonstrates that epidermal spheroid re-plating can enrich heterogenous normal human keratinocytes (NHKc) cultures for integrin&#945;6hi/EGFRlo keratinocyte subpopulations with enhanced stem-like characteristics. Our report describes the development and maintenance of a high throughput system for the study of skin keratinocyte plasticity and epidermal regeneration.

Here we describe a protocol for the systematic cultivation of epidermal spheroids in 3D suspension culture. This protocol has wide-ranging applications for use in a variety of epithelial tissue types and for the modeling of several human diseases and conditions.

建立高通量表皮球体培养系统，以模拟角膜细胞干细胞可塑性

Epithelial dysregulation is a node for a variety of human conditions and ailments, including chronic wounding, inflammation, and over 80% of all human ...

发育生物学

Generation of Tissue Spheroids via a 3D Printed Stamp-Like Device

Advances in 3D cell culture have developed more physiologically relevant in vitro models, such as tissue spheroids. Cells cultivated as spheroids have more realistic biological responses that resemble the in vivo environment. Due to their advantages, tissue spheroids represent an emerging trend toward superior, more reliable, and more predictive study models with a broad range of biotechnological applicability. However, reproducible platforms that can achieve large-scale production of tissue spheroids have become an unmet need in fully exploring and boosting their potential. Herein, the large-scale production of homogeneous tissue spheroids is reported using a low-cost and time-effective methodology. A 3D printed stamp-like device is developed to generate up to 4,716 spheroids per 6-well plate. The device is fabricated by the stereolithography method using a photocurable resin. The final device is composed of cylindrical micropins, with a height of 1.3 mm and a width of 650 &#181;m. This approach allows the fast generation of homogeneous spheroids and co-cultured spheroids with uniform shape and size and &gt;95% cell viability. Moreover, the stamp-like device is tunable for different sizes of well plates and Petri dishes. It is easily sterilized and can be reused for long periods. The efficient large-scale production of homogeneous tissue spheroids is essential to leverage their translation for multiple areas of industry, such as tissue engineering, drug development, disease modeling, and on-demand personalized medicine.

<meta charset="utf8"></head><body>通过 3D 打印的邮票状设备生成组织球体

The present protocol describes a technique to produce tissue spheroids on a large scale cost-effectively using a 3D printed stamp-like device.

通过 3D 打印的邮票状设备生成组织球体

Advances in 3D cell culture have developed more physiologically relevant in vitro models, such as tissue spheroids. Cells cultivated as spheroids have ...

生物化学

Semi-Automated Phenotypic Analysis of Functional 3D Spheroid Cell Cultures

We present a protocol that describes the properties and advantages of using a standalone clinostat incubator for growing, treating, and monitoring 3D cell cultures. The clinostat mimics an environment where cells can assemble as highly reproducible spheroids with low shear forces and active nutrient diffusion. We demonstrate that both cancer and non-cancer hepatocytes (HepG2/C3A and THLE-3 cell lines) require 3 weeks of growth prior to achieving functionalities comparable to liver cells. This protocol highlights the convenience of utilizing incubators for 3D cells with cameras monitoring the cell growth, as snapshots can be taken to count and measure spheroids upon treatment. We describe the comparison of THLE-3 and HepG2/C3A cell lines, showing how non-cancerous cell lines can be grown as well as immortalized cancer cells. We demonstrate and illustrate how proteomics experiments can be conducted from a few spheroids, which can be collected without perturbing cell signaling, i.e., no trypsinization required. We show that proteomics analysis can be used to monitor the typical liver phenotype of respiratory chain metabolism and the production of proteins involved in metal detoxification and describe a semi-automated system to count and measure the spheroid's area. Altogether, the protocol presents a toolbox that comprises a phenotypic characterization via image capture and a proteomics pipeline to experiment on 3D cell culture models.

We present a protocol for growing high-reproducible spheroids and their phenotypic characterization using image capture and proteomics.

功能性 3D 球状体细胞培养物的半自动表型分析

We present a protocol that describes the properties and advantages of using a standalone clinostat incubator for growing, treating, and monitoring 3D cell ...

用于基于 FLIM 的代谢和缺氧分析的多细胞球状体形成

Production and Multi-Parameter Live Cell Fluorescence Lifetime Imaging Microscopy (FLIM) of Multicellular Spheroids

Multicellular tumor spheroids are a popular 3D tissue microaggregate&#160;model for reproducing tumor microenvironment, testing and optimizing drug therapies&#160;and using bio- and nanosensors in a 3D context. Their ease of production, predictable size, growth, and observed nutrient and metabolite gradients are important to recapitulate the 3D niche-like&#160;cell microenvironment. However, spheroid heterogeneity and variability of their production methods can influence overall cell metabolism, viability, and drug response. This makes it difficult to choose the most appropriate&#160;methodology, considering the requirements in size, variability, needs of biofabrication, and use as in vitro 3D tissue models in stem and cancer cell biology. In particular, spheroid production can influence their compatibility with quantitative live microscopies, such as optical metabolic imaging, fluorescence lifetime imaging microscopy (FLIM), monitoring of spheroid&#160;hypoxia with nanosensors, or viability. Here, a number of conventional spheroid formation protocols are presented, highlighting their compatibility with the live widefield, confocal, and two-photon microscopies. The follow-up imaging to analysis&#160;pipeline with multiplexed autofluorescence FLIM and, using various types of cancer and stem cell spheroids, is also presented.

作者聚焦:使用荧光寿命成像显微镜标准化用于代谢和氧合分析的球状体形成方法

Here, we describe different multicellular spheroid formation methods to perform follow-up multi-parameter live cell microscopy. Using fluorescence lifetime imaging microscopy (FLIM), cellular autofluorescence, staining dyes, and nanoparticles, the approach for analysis of cell metabolism, hypoxia, and cell death in live three-dimensional (3D) cancer and stem cell-derived spheroids is demonstrated.

多细胞球体的生产和多参数活细胞荧光寿命成像显微镜 （FLIM）

Multicellular tumor spheroids are a popular 3D tissue microaggregate&#160;model for reproducing tumor microenvironment, testing and optimizing drug ...

蒸发减少培养条件的提高多细胞球体形成的微孔板重现

摘要

探索更多视频

此视频中的章节

生产尺寸控制肿瘤细胞球体采用微孔板大数

评估癌细胞3D球形培养中的细胞活力和死亡

利用细胞外通量分析探索单个3D微组织球体的线粒体能量代谢

建立高通量表皮球体培养系统，以模拟角膜细胞干细胞可塑性

通过 3D 打印的邮票状设备生成组织球体

通过 3D 打印的邮票状设备生成组织球体

功能性 3D 球状体细胞培养物的半自动表型分析

多细胞球体的生产和多参数活细胞荧光寿命成像显微镜（FLIM）

多细胞球体的生产和多参数活细胞荧光寿命成像显微镜（FLIM）

多细胞球体的生产和多参数活细胞荧光寿命成像显微镜（FLIM）

蒸发减少培养条件的提高多细胞球体形成的微孔板重现

摘要

探索更多视频

此视频中的章节

生产尺寸控制肿瘤细胞球体采用微孔板大数

评估癌细胞3D球形培养中的细胞活力和死亡

利用细胞外通量分析探索单个3D微组织球体的线粒体能量代谢

建立高通量表皮球体培养系统，以模拟角膜细胞干细胞可塑性

通过 3D 打印的邮票状设备生成组织球体

通过 3D 打印的邮票状设备生成组织球体

功能性 3D 球状体细胞培养物的半自动表型分析

多细胞球体的生产和多参数活细胞荧光寿命成像显微镜 （FLIM）

多细胞球体的生产和多参数活细胞荧光寿命成像显微镜 （FLIM）

多细胞球体的生产和多参数活细胞荧光寿命成像显微镜 （FLIM）

多细胞球体的生产和多参数活细胞荧光寿命成像显微镜（FLIM）

多细胞球体的生产和多参数活细胞荧光寿命成像显微镜（FLIM）

多细胞球体的生产和多参数活细胞荧光寿命成像显微镜（FLIM）