NEW YORK – Researchers from St. Jude Children’s Research Hospital, the University of Adelaide in Australia, and the National Center for Genomic Analysis (CNAG) in Spain are looking to spur the adoption of an imaging-based single-cell profiling method that promises to enable scalable and cost-effective profiling of millions of cells without the need for sequencing.

纽约——来自圣犹大儿童研究医院、澳大利亚阿德莱德大学和西班牙国家基因组分析中心 (CNAG) 的研究人员正致力于推动一种基于成像的单细胞分析方法的应用，这种方法有望在无需测序的情况下实现数百万个细胞的可扩展且经济高效的分析。

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The method, dubbed single-cell transcriptomics analysis and multimodal profiling through imaging (STAMP), was published in a paper in

该方法被称为单细胞转录组学分析和多模态成像分析（STAMP），发表在一篇论文中。

Cell

细胞

on Tuesday after being

周二在被

first described

首次描述

in a preprint in

在预印本中

BioRxiv

生物预印本

last year.

去年。

'There has been a lot of interest in the technology,' said Jasmine Plummer, a single-cell and spatial biology expert at St. Jude and one of the corresponding authors of the paper. 'Licensing can help get the method into a place where it would be replicable for other people.'

“这项技术引起了广泛关注，”圣犹大医院的单细胞与空间生物学专家、论文通讯作者之一贾斯明·普卢默表示。“通过授权，可以将这种方法推广到其他人能够复制应用的程度。”

A co-inventor of STAMP, Plummer said the team has filed a patent application for the technology in the US that was led by St Jude, with support from the University of Adelaide, the home institution of the technology's other co-inventor, Luciano Martelotto.

STAMP的共同发明人普拉默表示，该团队已经在美国为这项技术提交了专利申请，该申请由圣裘德牵头，阿德莱德大学（该技术另一位共同发明人卢西亚诺·马特洛托所在的机构）提供支持。

Plummer said that 'several companies' have reached out to St. Jude with interest for licensing the technology, though she declined to disclose their names.

普卢默说，“几家公司”已经与圣裘德接洽，表示有兴趣获得该技术的许可，不过她拒绝透露这些公司的名称。

The STAMP workflow begins with the fixation and permeabilization of cells in suspension, followed by adhering the cells onto glass slides to form monolayers, a step dubbed stamping. After stamping, the cells are incubated with gene probes or oligo-conjugated antibodies, followed by cyclic decoding through commercially available imaging platforms.

STAMP工作流程始于对悬浮细胞的固定和透化，随后将细胞贴附在玻片上形成单层，这一过程被称为“盖印”。盖印后，细胞与基因探针或寡核苷酸偶联抗体一起孵育，然后通过商业成像平台进行循环解码。

STAMP is flexible, given the adaptable format of stamping areas, the study authors noted, which allows for multi-sample profiling..

STAMP 是灵活的，研究作者指出，由于其可调适的印迹区域格式，允许进行多样本分析。

The researchers tested the method on diverse sample types, including peripheral blood mononuclear cells (PBMCs), dissociated cancer cells, and embryonic stem cell cultures, as well as whole cells and nuclei, and concluded that STAMP enables high-resolution single-cell RNA and protein profiling with great flexibility, throughput, and scale..

研究人员在多种样本类型上测试了该方法，包括外周血单核细胞 (PBMC)、分离的癌细胞、胚胎干细胞培养物，以及完整细胞和细胞核，并得出结论：STAMP 能够以极大的灵活性、通量和规模实现高分辨率的单细胞 RNA 和蛋白质分析。

In the

在

Cell

细胞

paper, they demonstrated the method's compatibility with four imaging platforms: the 10x Genomics Xenium Analyzer, the Bruker Spatial Biology CosMx Spatial Molecular Imager, the Merscope platform from Vizgen, and the Akoya Biosciences PhenoCycler-Fusion instrument. 'The whole purpose of STAMP is to be instrument-agnostic,' Plummer said.

在论文中，他们展示了该方法与四个成像平台的兼容性：10x Genomics 的 Xenium 分析仪、Bruker 空间生物学 CosMx 空间分子成像仪、Vizgen 的 Merscope 平台以及 Akoya Biosciences 的 PhenoCycler-Fusion 仪器。普卢默表示：“STAMP 的全部意义在于不受仪器限制。”

'This just shows the method can go on any surface area.' .

“这仅仅表明该方法可以应用于任何表面区域。”

While the preprint focused on cell line samples, in the

虽然预印本专注于细胞系样本，但ใน

Cell

细胞

paper, the researchers further assessed the applicability of STAMP for dissociated tissues, analyzing samples from five major mouse organs: kidney, liver, brain, heart, and lung. The results indicated that the method is reliable and reproducible in single cells from such tissue samples.

论文中，研究人员进一步评估了STAMP对于分离组织的适用性，分析了来自小鼠五个主要器官（肾脏、肝脏、大脑、心脏和肺）的样本。结果表明，该方法在来自此类组织样本的单细胞中是可靠且可重复的。

They also demonstrated multiomics profiling of RNA and protein with STAMP. In one experiment, they profiled the RNA of the stamped cells using the Xenium platform, then reused the same slides for subsequent protein profiling with two separate platforms, the CosMx SMI and the PhenoCycler-Fusion.

他们还展示了使用STAMP进行RNA和蛋白质的多组学分析。在一项实验中，他们使用Xenium平台对标记细胞的RNA进行了分析，然后将相同的玻片用于后续的蛋白质分析，采用了两个独立的平台：CosMx SMI和PhenoCycler-Fusion。

The protein assays produced high-quality signals, the authors noted, illustrating that prior transcriptome analysis did not notably impact protein profiling.

作者指出，蛋白质检测产生了高质量的信号，这表明之前的转录组分析并未显著影响蛋白质谱分析。

'That means those cells stay on [the STAMP slides] long enough to move it from machine to machine,' Plummer said. 'You can do your assay, leave it in the fridge, and with the right conditions, get your RNA off and put it onto another machine — that's pretty amazing and robust.'

“这意味着这些细胞会停留在[STAMP载玻片]上足够长的时间，以便在不同设备之间移动，”普卢默说。“你可以进行检测，然后把它放在冰箱里，在合适的条件下，获取你的RNA并将其转移到另一台设备上——这非常令人惊叹且十分可靠。”

The stability of the STAMP samples also enables researchers to ship slides to different labs with different equipment for collaboration and analysis, she added.

她补充说，STAMP 样品的稳定性还使研究人员能够将玻片运送到配备不同设备的不同实验室，以进行合作和分析。

Besides being able to analyze millions of cells, the researchers also demonstrated STAMP's sensitivity for detecting rare cell types by spiking cancer cells at 1:100,000 and 1:50,000 dilutions into PBMC samples. As such, STAMP could also potentially be used for clinically relevant scenarios, such as detecting circulating tumor cells, they noted.

除了能够分析数百万个细胞外，研究人员还通过将癌细胞以1:100,000和1:50,000的比例稀释到PBMC样本中，展示了STAMP检测稀有细胞类型的灵敏度。他们指出，因此STAMP也可能用于临床相关场景，例如检测循环肿瘤细胞。

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'So many times in single-cell studies, we don't have enough cells,' Plummer noted. '[This experiment] was intentional on our part as researchers, because for us, we are trying to get this assay to work for very low numbers of cells.'

“在单细胞研究中，很多时候我们没有足够的细胞，”普卢默指出。“[这个实验]是我们研究人员有意为之，因为对我们来说，我们正试图让这种检测方法在细胞数量非常少的情况下发挥作用。”

Given that STAMP works without sequencing, the authors also touted its low cost. By their account, analyzing 1,000 PBMC samples at 25,000 cells per sample, for instance, would cost around $3.6 million using single-cell sequencing, whereas the cost for STAMP is $75,000. They estimated that the cost for single-cell sequencing is approximately $.14 per cell, compared to a cost of $.003 per cell for STAMP.

鉴于 STAMP 无需测序即可工作，作者还大肆宣扬其低成本。据他们估算，例如，分析 1,000 个 PBMC 样本，每个样本 25,000 个细胞，使用单细胞测序的成本约为 360 万美元，而 STAMP 的成本为 75,000 美元。他们估计，单细胞测序的成本约为每个细胞 0.14 美元，而 STAMP 每个细胞的成本为 0.003 美元。

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Despite these promises, the method has limitations. Since it relies on predefined gene panels, for instance, it cannot discover novel transcripts. Additionally, it uses probe-based detection, which can introduce hybridization biases, potentially skewing the results, the study authors noted.

尽管有这些优势，该方法仍存在局限性。例如，由于该方法依赖于预定义的基因面板，因此无法发现新的转录本。此外，它使用基于探针的检测方法，这可能会引入杂交偏差，从而可能扭曲结果，研究作者指出。

Without a commercialized product, it also remains to be seen how easily researchers can adopt the method.

没有商业化的产品，研究人员能否轻松采用该方法还有待观察。

'I do know that there are nuances to it,' Plummer said. 'We have been running single-cell [experiments] for over 10 years, so knowing all the tricks and trades of molecular biology will help.' A commercial STAMP kit could help foster the adoption of the method, she added.

“我确实知道这其中有一些细微之处，”普卢默说。“我们已经进行了超过10年的单细胞[实验]，所以了解分子生物学的所有技巧和方法会有帮助。”她补充说，一个商用的STAMP试剂盒可能有助于促进该方法的采用。

As for applications, Plummer said her lab is currently using STAMP to streamline CRISPR Perturb-seq experiments for drug discovery.

至于应用，普卢默说她的实验室目前正在使用STAMP来简化用于药物发现的CRISPR Perturb-seq实验。

'One of the overwhelming things for the Perturb-seq assay is that there is a lot of cloning involved, and a lot of sequencing to find that individual single clone. It is really laborious to do,' said Plummer, adding that with STAMP, they can now process a large number of cells without sequencing.

“Perturb-seq检测的一个艰巨之处在于，它涉及大量的克隆工作，并且需要进行大量的测序来找到单个单克隆。这个过程非常费力。”普卢默补充说，使用STAMP技术，他们现在可以处理大量细胞而无需进行测序。

Given STAMP is a slide-based approach, the method, which retains cellular structure and morphology, also enables hematoxylin and eosin (H&E) staining, a gold standard in pathology, making it compatible with clinical applications, Plummer said.

鉴于STAMP是一种基于幻灯片的方法，普卢默说，该方法保留了细胞结构和形态，还能够进行苏木精和伊红（H&E）染色这一病理学的金标准，使其适用于临床应用。

'Most clinical diagnostics, especially for solid tissue, are done on a slide,' Plummer said. 'Now, any discovery that we do, we have kind of put [STAMP and clinical pathology] in the same universe.'

“大多数临床诊断，尤其是实体组织的诊断，都是在玻片上完成的，”普卢默说。“现在，我们所做的任何发现，都可以说是将[STAMP和临床病理学]放在了同一个领域。”