Molecular mapping

分子作图
  • 文章类型: Journal Article
    果皮颜色是影响水果品质和顾客需求的主要因素。然而,西葫芦绿色果皮颜色性状的分子机制尚不清楚。两条家长线,RP16和RP38用于研究C.pepo的果皮颜色性状。亲本系RP16显示白色果皮颜色,而RP38表现出绿色的果皮颜色。384个F2种群用于鉴定南瓜中与绿色果实和白色果皮相关的遗传模式。293个F2个体为白色,91个F2个体是绿色的,导致比例为3:1。因此,白色果皮在绿色果皮性状上占主导地位,和单个隐性绿皮基因(Cpgp)控制绿色果皮。在RP16品系中,随着果实的成熟,果实叶绿素(Chll)含量降低。相比之下,在RP38品系的果皮上,Chll在果实生长期增加。BSA序列分析揭示了Chr5上2.3Mb区域内的Cpgp基因座。随后的精细映射分析,使用699个F2植物,将该区域缩小到同一染色体上的23.90kb。在这个区域内,两个注释基因,即Cp4.1LG05g02070和Cp4.1LG05g02060存在。预测这些基因编码两组分拟南芥假反应调节因子2样蛋白(APRR2),这可能与植物的绿色色素沉着过程有关。因此,在各个果实发育阶段的序列比对和基因表达分析支持Cp4.1LG05g02070可能是负责调节葫芦绿色果皮颜色性状的主要候选基因。这项研究可能为进一步研究控制南瓜果皮颜色的基本机制提供基础。
    在线版本包含补充材料,可在10.1007/s11032-024-01492-7获得。
    Fruit peel color is a major factor that influences fruit quality and customers\' demand. However, the molecular mechanisms underlying the green fruit peel color trait of Cucurbita pepo L. remain unknown. Two parental lines, RP16 and RP38, were used to study the fruit peel color trait in C. pepo. The parental line RP16 shows white peel color, whereas RP38 exhibits green peel color. 384 F2 populations were used to identify the inheritance pattern associated with green fruit and white fruit peel in Cucurbita pepo L. 293 F2 individuals were white, and 91 F2 individuals were green, resulting in a ratio of 3:1. Hence, white peel is dominant over the green fruit peel trait, and a single recessive green peel gene (Cpgp) controls the green fruit peel. The fruit chlorophyll (Chll) content decreases as fruit matures in the RP16 line. In contrast, Chll increases during the fruit growing periods on fruit peels of the RP38 line. The BSA-sequence analysis revealed the Cpgp locus on Chr5, within a 2.3 Mb region. Subsequent fine-mapping analysis, using 699 F2 plants, narrowed down this region to 23.90 kb on the same chromosome. Within this region, two annotated genes, namely Cp4.1LG05g02070 and Cp4.1LG05g02060, are present. These genes are predicted to encode a two-component Arabidopsis Pseudo-Response Regulator 2-like protein (APRR2), which may be involved in green pigmentation processes in plants. Consequently, sequence alignment and gene expression analyses at various fruit development stages supported that Cp4.1LG05g02070 may be the primary candidate gene responsible for regulating the green fruit peel color trait in Cucurbita pepo L. This study may provide a basis for further study on the basic mechanisms that control the fruit peel colors in Cucurbita spp.
    UNASSIGNED: The online version contains supplementary material available at 10.1007/s11032-024-01492-7.
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  • 文章类型: Journal Article
    小麦(TriticumaestivumL.)是世界上最重要的农作物之一。由Blumeriagraminisf.sp.引起的白粉病。小麦(Bgt)是威胁小麦产量和品质的破坏性疾病。抗性基因和品种的利用被认为是最经济的,环保,控制白粉病的有效方法。小麦育种系京子102在苗期和成株期均对白粉病具有很强的抗性。F1,F2,F2的遗传分析:\"京子102×世新828\"3个种群表明,京子102苗期对白粉病分离株E09的抗性受单一显性基因控制,临时指定PmJZ。使用块状分离体RNA-Seq结合分子标记分析,PmJZ位于2B染色体的长臂上,两侧有标记BJK695-1和CIT02g-20,遗传距离为1.2和0.5cM,分别,对应于中国春季的面包小麦基因组(IWGSCRefSeqv2.1)703.8-707.6Mb。PmJZ根据其物理位置很可能与2BL染色体上记录的Pm基因不同,分子标记分析,和抗性谱。基于基因注释信息,5个与抗病性相关的基因可作为PmJZ的候选基因。为了加快PmJZ的应用,侧翼标记BJK695-1和CIT02g-20可用于小麦抗病育种中PmJZ的标记辅助选择。
    Wheat (Triticum aestivum L.) is one of the most important crops worldwide. Powdery mildew caused by Blumeria graminis f. sp. tritici (Bgt) is a destructive disease threatening wheat yield and quality. The utilization of resistant genes and cultivars is considered the most economical, environmentally-friendly, and effective method to control powdery mildew. Wheat breeding line Jingzi 102 was highly resistant to powdery mildew at both seedling and adult plant stages. Genetic analysis of F1, F2, and F2:3 populations of \"Jingzi 102 × Shixin 828\" showed that the resistance of Jingzi 102 against powdery mildew isolate E09 at the seedling stage was controlled by a single dominant gene, temporarily designated PmJZ. Using bulked segregant RNA-Seq combined with molecular markers analysis, PmJZ was located on the long arm of chromosome 2B and flanked by markers BJK695-1 and CIT02g-20 with the genetic distances of 1.2 and 0.5 cM, respectively, corresponding to the bread wheat genome of Chinese Spring (IWGSC RefSeq v2.1) 703.8-707.6 Mb. PmJZ is most likely different from the documented Pm genes on chromosome 2BL based on their physical positions, molecular markers analysis, and resistance spectrum. Based on the gene annotation information, five genes related to disease resistance could be considered as the candidate genes of PmJZ. To accelerate the application of PmJZ, the flanking markers BJK695-1 and CIT02g-20 can serve for marker-assisted selection of PmJZ in wheat disease resistance breeding.
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  • 文章类型: Journal Article
    小麦白粉病,由生物营养真菌Blumeriagraminisf.sp.小麦(Bgt),是影响全世界小麦的最具破坏性的疾病之一。选育抗病小麦品种是防治该病最经济有效的方法之一,因此,识别和定位新的和有效的抗性基因是至关重要的。Baidatou,一种中国小麦地方品种,表现出优异的抗白粉病性。为了鉴定白大头的抗性基因,分别于2019年、2020年和2021年在陕西杨凌(YL)和甘肃天水(TS)试验田中对来自明县169/白达头杂交的170个F7:8个重组自交系(RILs)进行成株期白粉病响应评价。明县169/白大头F7:8RIL的疾病进展曲线下相对面积(rAUDPC)表明,白大头对白粉病的抗性受数量性状位点(QTLs)控制。基于批量分离分析,结合660K单核苷酸多态性(SNP)阵列和通过整个RIL群体的靶测序(16KSNP)进行基因分型,两个QTL,QPmbdt。nwafu-2AS和QPmbdt。nwafu-3AS,被确认,这些占表型变异的44.5%。其中一个QTL位于竞争性等位基因特异性PCR标记AX-111012288和AX_174233809之间的小麦2AS染色体上的3.32cM遗传区间上,另一个位于SNP标记3A_684044820和3A_686681822之间的3AS染色体上的9.6cM遗传区间上。这些标记可用于小麦白粉病抗性的成功育种。
    Wheat powdery mildew, caused by the biotrophic fungus Blumeria graminis f. sp. tritici (Bgt), is one of the most devastating diseases affecting wheat throughout the world. Breeding and growing resistant wheat cultivars is one of the most economic and effective methods to control the disease, and as such, identifying and mapping the new and effective resistance genes is critical. Baidatou, a Chinese wheat landrace, shows excellent field resistance to powdery mildew. To identify the resistance gene(s) in Baidatou, 170 F7:8 recombinant inbred lines (RILs) derived from the cross Mingxian 169/Baidatou were evaluated for powdery mildew response at the adult-plant stage in the experimental fields in Yangling (YL) of Shaanxi Province and Tianshui (TS) in Gansu Province in 2019, 2020, and 2021. The relative area under disease progress curve (rAUDPC) of Mingxian 169/Baidatou F7:8 RILs indicated that the resistance of Baidatou to powdery mildew was controlled by quantitative trait loci (QTLs). Based on bulk segregation analysis combined with the 660K single nucleotide polymorphism (SNP) array and genotyping by target sequencing (16K SNP) of the entire RIL population, two QTLs, QPmbdt.nwafu-2AS and QPmbdt.nwafu-3AS, were identified, and these accounted for up to 44.5% of the phenotypic variation. One of the QTLs was located on the 3.32 cM genetic interval on wheat chromosome 2AS between the kompetitive allele-specific PCR markers AX-111012288 and AX_174233809, and another was located on the 9.6 cM genetic interval on chromosome 3AS between the SNP markers 3A_684044820 and 3A_686681822. These markers could be useful for successful breeding of powdery mildew resistance in wheat.
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  • 文章类型: Journal Article
    Centriole生物发生,和大多数细胞器装配一样,涉及顺序招募将支持其功能的子结构要素。为了揭开这个过程,我们使用扩展显微镜将24个中心摩尔蛋白的空间位置与结构特征相关联。整个人类原粒组装过程中蛋白质分布的时间序列重建揭示了中心粒生物生成步骤的分子结构。我们发现该过程始于没有微管的裸露车轮的形成。接下来,开花阶段随着微管叶片组装而进展,伴随着径向分离和快速的车轮生长。在随后的延伸阶段,微管蛋白骨架随着A-C接头的募集线性增长,其次是内部支架(IS)的蛋白质。通过遵循六个结构模块,我们模拟了人体中心的4D组装。总的来说,这项工作为研究大型大分子的时空组装提供了一个框架。
    Centriole biogenesis, as in most organelle assemblies, involves the sequential recruitment of sub-structural elements that will support its function. To uncover this process, we correlated the spatial location of 24 centriolar proteins with structural features using expansion microscopy. A time-series reconstruction of protein distributions throughout human procentriole assembly unveiled the molecular architecture of the centriole biogenesis steps. We found that the process initiates with the formation of a naked cartwheel devoid of microtubules. Next, the bloom phase progresses with microtubule blade assembly, concomitantly with radial separation and rapid cartwheel growth. In the subsequent elongation phase, the tubulin backbone grows linearly with the recruitment of the A-C linker, followed by proteins of the inner scaffold (IS). By following six structural modules, we modeled 4D assembly of the human centriole. Collectively, this work provides a framework to investigate the spatial and temporal assembly of large macromolecules.
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  • 文章类型: Journal Article
    白粉病,由Blumeriagraminisf.sp.小麦(Bgt),是一种严重的疾病,影响小麦的产量和品质。在生产中推广抗性品种是控制该疾病的首选策略。在本研究中,中国小麦育种系吉麦809在整个生长阶段表现出优异的农艺性能和对白粉病的高抗性。为了剖析这种抗性的遗传基础,将Jimai809与易感小麦品种Junda159杂交,以产生隔离种群。遗传分析表明,单个显性基因,暂时命名为PmJM809,赋予不同的Bgt分离株抗性。然后将PmJM809定位在染色体臂2BL上,并由标记器CISSR02g-1和CIT02g-13侧翼,遗传距离为0.4和0.8cM,分别,对应于704.12-708.24Mb的物理间隔。PmJM809与所报道的染色体臂2BL上的Pm基因的起源不同,电阻谱,绘图间隔的物理位置和/或遗传多样性,这也表明PmJM809位于具有多个抗性基因的复杂区间。为了分析和筛选PmJM809的候选基因,使用另外的一组小麦样品和接种Bgt分离株E09后的时程分析评估候选区间中与抗病性相关的六个基因的表达模式。因此,四个基因被推测为关键候选或调节基因。综合农艺性状和抗性研究结果,PmJM809有望成为小麦抗病育种的宝贵基因资源。为了有效地将PmJM809转移到不同的遗传背景中,19个紧密连锁的标记中的13个被证实适用于标记辅助选择。使用这些标记,从吉麦809和几个易感品种的杂交中选择了一系列具有协调抗病性和农艺性能的小麦育种系。
    在线版本包含补充材料,可在10.1007/s11032-024-01467-8获得。
    Powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is a severe disease that affects the yield and quality of wheat. Popularization of resistant cultivars in production is the preferred strategy to control this disease. In the present study, the Chinese wheat breeding line Jimai 809 showed excellent agronomic performance and high resistance to powdery mildew at the whole growth stage. To dissect the genetic basis for this resistance, Jimai 809 was crossed with the susceptible wheat cultivar Junda 159 to produce segregation populations. Genetic analysis showed that a single dominant gene, temporarily designated PmJM809, conferred the resistance to different Bgt isolates. PmJM809 was then mapped on the chromosome arm 2BL and flanked by the markers CISSR02g-1 and CIT02g-13 with genetic distances 0.4 and 0.8 cM, respectively, corresponding to a physical interval of 704.12-708.24 Mb. PmJM809 differed from the reported Pm genes on chromosome arm 2BL in origin, resistance spectrum, physical position and/or genetic diversity of the mapping interval, also suggesting PmJM809 was located on a complex interval with multiple resistance genes. To analyze and screen the candidate gene(s) of PmJM809, six genes related to disease resistance in the candidate interval were evaluated their expression patterns using an additional set of wheat samples and time-course analysis post-inoculation of the Bgt isolate E09. As a result, four genes were speculated as the key candidate or regulatory genes. Considering its comprehensive agronomic traits and resistance findings, PmJM809 was expected to be a valuable gene resource in wheat disease resistance breeding. To efficiently transfer PmJM809 into different genetic backgrounds, 13 of 19 closely linked markers were confirmed to be suitable for marker-assisted selection. Using these markers, a series of wheat breeding lines with harmonious disease resistance and agronomic performance were selected from the crosses of Jimai 809 and several susceptible cultivars.
    UNASSIGNED: The online version contains supplementary material available at 10.1007/s11032-024-01467-8.
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  • 文章类型: Journal Article
    多年来,成像神经元结构一直是一个反复出现的挑战,突触蛋白的定位是神经科学中的一个常见挑战。为了定量检测和分析突触的结构,我们最近开发了免费的SODA软件来检测突触前和突触后蛋白的关联。为了充分利用复杂细胞中的空间分布分析,比如神经元,我们还选择了一些新的染料用于质膜标记。使用冰冷的SODA插件,我们可以在常规和单分子定位显微镜中检测和分析突触关联,获得纳米级分子图谱。为了取代神经元三维(3D)形状内的分子分布,我们使用MemBright探针和3DSTORM分析以单分子分辨率水平破译各种树突状脊柱类型的整个3D形状.我们在这里报告了神经元掩模中的突触蛋白的例子,但是这些工具具有更广泛的兴趣,因为它们可以用于任何蛋白质或细胞类型。连同SODA插件,因此,MemBright探针提供了完美的工具包来破译3D细胞环境中蛋白质的纳米分子图。
    Imaging neuronal architecture has been a recurrent challenge over the years, and the localization of synaptic proteins is a frequent challenge in neuroscience. To quantitatively detect and analyze the structure of synapses, we recently developed free SODA software to detect the association of pre and postsynaptic proteins. To fully take advantage of spatial distribution analysis in complex cells, such as neurons, we also selected some new dyes for plasma membrane labeling. Using Icy SODA plugin, we could detect and analyze synaptic association in both conventional and single molecule localization microscopy, giving access to a molecular map at the nanoscale level. To replace those molecular distributions within the neuronal three-dimensional (3D) shape, we used MemBright probes and 3D STORM analysis to decipher the entire 3D shape of various dendritic spine types at the single-molecule resolution level. We report here the example of synaptic proteins within neuronal mask, but these tools have a broader spectrum of interest since they can be used whatever the proteins or the cellular type. Altogether with SODA plugin, MemBright probes thus provide the perfect toolkit to decipher a nanometric molecular map of proteins within a 3D cellular context.
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  • 文章类型: Journal Article
    低温和冷害是严重降低小麦产量的自然因素。因此,如何提高小麦的抗寒性一直是小麦育种家和遗传学家关注的焦点。然而,这种性状的遗传改进一直很缓慢,主要是因为抗寒性是一个复杂的数量性状,田间表型鉴定相对困难。因此,发现,映射,小麦抗寒基因的克隆为小麦抗寒的遗传改良提供了理论依据,并有助于分析小麦抗寒的分子机制。本研究以小麦品系H261及其EMS突变体LF2099和XiNong239为材料。突变体LF2099和XiNong239的F2代分离以15:1的比例发生了冷性状分离。遗传分析表明,两个显性重叠基因,暂时命名为Wcr-3和Wcr-4,控制小麦的抗寒性。此外,组合的BSA和SNP阵列确定Wcr-3在BU100519(SSR标记)和AX-94843669(SNP标记)之间。标记相距1.32cM,对应于中国春季2B染色体上的5.41Mb物理间隔,具有67个功能注释基因。Wcr-4位于AX-94657955(SNP标记)和LC-23(SSR标记)之间,相距1.79cM,对应于中国春季2D染色体上的2.35Mb物理间隔,其中包含66个功能注释的基因。Wcr-3和Wcr-4是两个新的抗寒基因,为他们精细的作图和克隆奠定了基础。
    在线版本包含补充材料,可在10.1007/s11032-023-01425-w获得。
    Low temperature and cold damage are natural factors that seriously reduce wheat yield. Thus, how to improve the cold resistance of wheat has been the focus of wheat breeders and geneticists. However, the genetic improvement for this trait has been slow, mainly because cold resistance is a complex quantitative trait and field phenotypic identification is relatively difficult. Therefore, the discovery, mapping, and cloning of the cold resistance genes of wheat provide a theoretical basis for the genetic improvement of wheat against cold resistance and facilitate the analysis of the molecular mechanisms of cold resistance in wheat. This study used the wheat line H261 and its EMS mutants LF2099 and XiNong 239 as materials. Cold trait segregation occurred in the F2 generation of mutants LF2099 and XiNong 239 at a 15:1 separation ratio. Genetic analysis showed that two dominant overlapping genes, temporarily named Wcr-3 and Wcr-4, control cold resistance in wheat. Furthermore, a combined BSA and SNP array established that Wcr-3 is between BU100519 (SSR marker) and AX-94843669 (SNP marker). The markers are 1.32 cM apart, corresponding to the 5.41 Mb physical interval on the Chinese Spring 2B chromosome with 67 functionally annotated genes. Wcr-4 is located between AX-94657955 (SNP marker) and LC-23 (SSR marker), which are 1.79 cM apart, corresponding to a 2.35 Mb physical interval on the Chinese Spring 2D chromosome, which contains 66 functionally annotated genes. Wcr-3 and Wcr-4 are two new cold resistance genes, laying the foundation for their fine mapping and cloning.
    UNASSIGNED: The online version contains supplementary material available at 10.1007/s11032-023-01425-w.
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  • 文章类型: Journal Article
    小麦叶锈病(Lr),是由PucciniatriticinaEriks引起的.(Pt)是影响全球小麦生产的最重要的小麦病害之一。使用抗性小麦品种是控制叶锈病最经济,最环保的方法。意大利小麦品种Libellula在田间研究中表现出对Lr的良好抗性。为了确定Libellula中Lr抗性的遗传基础,来自Libellula/Huixianhong杂交的248F6重组自交系在七个环境中进行了Lr严重程度的表型分析:保定的2014/2015、2016/2017、2017/2018和2018/2019种植季节,河北省和周口的2016/2017、2017/2018和2018/2019作物季节,河南省。然后使用大量的分离分析和简单的序列重复标记来鉴定种群中Lr成年植物抗性的定量性状基因座(QTL)。因此检测到六个QTL,指定为QLr。hebau-1AL和QLr。hebau-1AS假定是新的,QLr。hebau-1BL,QLr。hebau-3AL,QLr。hebau-4BL,和QLr。hebau-7DS在与先前报道的QTL相似的物理位置识别。根据染色体位置和分子标记测试,QLr。hebau-1BL和QLr。hebau-7DS分别与Lr46和Lr34具有相似的侧翼标记。Lr46和Lr34是叶锈病的种族非特异性APR基因,条锈病,和白粉病.同样,QLr。Hebau-4BL对叶锈病表现出多种抗病性,条锈病,镰刀菌枯萎病,和白粉病.本研究中确定的QTL,以及它们紧密相连的标记,有可能用于小麦育种中的标记辅助选择。
    Wheat leaf rust (Lr), which is caused by Puccinia triticina Eriks. (Pt), is one of the most important wheat diseases affecting wheat production globally. Using resistant wheat cultivars is the most economical and environmentally friendly way to control leaf rust. The Italian wheat cultivar Libellula has demonstrated good resistance to Lr in field studies. To identify the genetic basis of Lr resistance in \'Libellula\', 248 F6 recombinant inbred lines from the cross \'Libellula\'/\'Huixianhong\' was phenotyped for Lr severity in seven environments: the 2014/2015, 2016/2017, 2017/2018, and 2018/2019 cropping seasons at Baoding, Hebei Province, and the 2016/2017, 2017/2018, and 2018/2019 crop seasons at Zhoukou, Henan Province. Bulked segregant analysis and simple sequence repeat markers were then used to identify the quantitative trait loci (QTLs) for Lr adult-plant resistance in the population. Six QTLs were consequently detected and designated as QLr.hebau-1AL and QLr.hebau-1AS that were presumed to be new and QLr.hebau-1BL, QLr.hebau-3AL, QLr.hebau-4BL, and QLr.hebau-7DS that were identified at similar physical positions as previously reported QTLs. Based on chromosome positions and molecular marker tests, QLr.hebau-1BL and QLr.hebau-7DS share similar flanking markers with Lr46 and Lr34, respectively. Lr46 and Lr34 are race nonspecific adult plant resistance (APR) genes for leaf rust and stripe rust and powdery mildew. QLr.hebau-4BL showed multiple disease resistance to leaf rust, stripe rust, Fusarium head blight, and powdery mildew. The QTL identified in this study, as well as their closely linked markers, may potentially be used in marker-assisted selection in wheat breeding.
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  • 文章类型: Journal Article
    质谱成像(MSI)是一种灵敏的,具体,无标签成像分析技术,可以同时获得空间分布,相对含量,以及细胞和组织中数百种生物分子的结构信息,如脂质,药物小分子,肽,蛋白质,和其他化合物。单细胞分子图谱的研究可以揭示生物体的活动模式等重大科学问题,疾病的发病机理,药物靶向治疗,和细胞异质性。将MSI技术应用于单细胞分子图谱可以为单细胞代谢组学的研究提供新的见解和思路。这篇综述旨在为MSI社区中对单细胞成像感兴趣的人提供信息资源。特别是,我们讨论成像方案和样品制备的进展,仪表改进,数据处理和分析,和3DMSI在过去几年中,使MSI成为单细胞分子成像中的强大技术。此外,我们强调了单细胞MSI中一些最尖端的研究,展示了单细胞MSI的未来潜力。在单细胞甚至亚细胞水平上可视化分子分布可以为我们提供更丰富的细胞信息,这极大地促进了生物医学等研究领域的发展,生命科学,药效学试验,和代谢组学。在审查结束时,我们总结了目前单细胞MSI技术的发展现状,并展望了该技术的未来。
    Mass spectrometry imaging (MSI) is a sensitive, specific, label-free imaging analysis technique that can simultaneously obtain the spatial distribution, relative content, and structural information of hundreds of biomolecules in cells and tissues, such as lipids, small drug molecules, peptides, proteins, and other compounds. The study of molecular mapping of single cells can reveal major scientific issues such as the activity pattern of living organisms, disease pathogenesis, drug-targeted therapy, and cellular heterogeneity. Applying MSI technology to the molecular mapping of single cells can provide new insights and ideas for the study of single-cell metabolomics. This review aims to provide an informative resource for those in the MSI community who are interested in single-cell imaging. Particularly, we discuss advances in imaging schemes and sample preparation, instrumentation improvements, data processing and analysis, and 3D MSI over the past few years that have allowed MSI to emerge as a powerful technique in the molecular imaging of single cells. Also, we highlight some of the most cutting-edge studies in single-cell MSI, demonstrating the future potential of single-cell MSI. Visualizing molecular distribution at the single-cell or even sub-cellular level can provide us with richer cell information, which strongly contributes to advancing research fields such as biomedicine, life sciences, pharmacodynamic testing, and metabolomics. At the end of the review, we summarize the current development of single-cell MSI technology and look into the future of this technology.
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  • 文章类型: Editorial
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