Resources:
固定,批量处理,灵活运行
Take advantage of robust fixation protocols that let you run assays on your schedule, simplify sample transport, and optimize your workflow.
分析新鲜、冷冻和固定样本
Easily analyze previously inaccessible samples, from fixed tissues (even FFPE) to cells, nuclei, and FACS-sorted cells.
降低每个样本的成本
批量和混样分析可使得每个样本的成本降低多达75%,并降低测序成本,最大限度利用您的资源。
Ultimate experimental flexibility
Process up to 1 million cells per run, change batch and sample throughput with ease, and capture protein alongside gene expression.
行业领先的性能表现
Be confident in your data, using unmatched sensitivity for whole transcriptome profiling to analyze even damaged RNA or low-expressing genes.
简化的工作流程和数据分析
您可以通过轻松且可重复的实验方案、简化的数据管理和快速处理来探索基因表达谱。
您可以做什么
简化样本的物流运输,并设置自己的实验日程表
样本固定和储存可以实现优化且灵活的样本批量处理,同时减少可变性并保持数据质量。
从挑战性的样本中获得高质量的数据,即使是FFPE样本
创新的探针设计带来了灵敏而可靠的性能,即使是整块组织、FFPE样本以及带有片段化RNA的样本。
将经济高效的scRNA-seq推广到更多样本
降低每个样本的成本,扩大单细胞实验的范围,让更大规模的实验得以开展。
轻松开展多地实验
可靠的操作方案可保留珍贵样本的生物学特征,让全球范围内的研究成为可能。
Capture the data that fits your research needs
Assay flexibility lets you capture protein alongside gene expression, or target specific RNAs with user-designed probe pairs to supplement the whole transcriptome panel.
经过验证的结果
视频
资源
Robust results for scRNA-seq from matched FFPE and freshly fixed samples
Robust results for scRNA-seq from matched FFPE and freshly fixed samples
Data Spotlight
Data Spotlight
Isolation of Cells from FFPE Tissue Sections for Chromium Fixed RNA Profiling
Isolation of Cells from FFPE Tissue Sections for Chromium Fixed RNA Profiling
Demonstrated Protocol
Demonstrated Protocol
Single Cell Gene Expression Flex: Lock in cell states, unlock potential
Single Cell Gene Expression Flex: Lock in cell states, unlock potential
Blog
Blog
完整的解决方案

Chromium单细胞基因表达Flex试剂盒
With our reagent kits, explore cellular diversity through single cell, whole transcriptome analysis of fresh, frozen, or fixed samples for human or mouse, including simultaneous measurement of cell surface proteins.
工作流程
Scroll to see the base workflow for Single Cell Gene Expression Flex. Options are included for sample multiplexing to increase throughput, or for use of Feature Barcode technology for cell surface protein detection in singleplex or multiplex workflows.
- 1
采集样本
Start with fresh, frozen, or fixed cells, nuclei, or whole tissue. 对于新鲜样本,请按照样本制备的最佳做法进行操作,以便获得高质量的单细胞悬液(或跳到第2步进行组织固定)。If desired, stain whole cells with Feature Barcode technology to profile cell surface proteins simultaneously with gene expression.
对于FFPE样本,请按照我们的示范方案进行操作,以便获得如何解离FFPE组织用于此分析的指引。After dissociation, samples can be safely stored at 4°C for up to 1 week or –80°C for up to 6 months. If not storing dissociated samples, you can instead move directly to Step 3.
- 2
样本固定和透化
按照我们简单明了的方案用多聚甲醛(PFA)固定样本,同时锁定生物学状态并保留脆弱细胞。此时,样本可在-80℃进行安全运输或储存,直到您准备好处理这些样本为止。
如果您处理的是FFPE样本,请跳过这一部分,直接开始第3步。
资源 - 3
构建10x文库
Hybridize the probe set to your fixed samples, then partition individual cells on the Chromium iX or X instrument. After ligation and extension reactions, construct a 10x barcoded library. 在每个纳升规模的分区反应体系内,细胞发生以RNA为模板的连接,因此每个由此生成的产物都共享一个来自其单个来源细胞的10x条形码。For multiplexing workflows to increase throughput, hybridize your samples to probe sets, pool samples together, and proceed as usual—with up to 16 samples in a single lane of a Chromium chip. 在这种情况下,附加在每组探针上的另一个独特条形码可帮助您将每个单独的分子分配回它来源的样本。
If, prior to sample fixation, whole cells were stained with Feature Barcode technology to profile cell surface proteins, a Protein Expression Library can be generated alongside the Gene Expression Library.
资源- Chromium Fixed RNA Profiling – Protocol Planner
- User Guide for Singleplex Samples
- User Guide for Singleplex Samples with Feature Barcode technology for Protein
- User Guide for Singleplex Samples with Feature Barcode technology for Protein using Barcode Oligo Capture
- 多个样本分析的操作指南
- User Guide for Multiplexed Samples with Feature Barcode technology for Protein using Barcode Oligo Capture
- 4
测序
Resulting 10x barcoded libraries are compatible with standard NGS short-read sequencing on Illumina sequencers for massive transcriptional profiling of hundreds of thousands of individual cells at once.
资源 - 5
数据分析和可视化
我们的Cell Ranger分析软件可生成每个细胞的基因表达谱,识别表达谱相似的细胞簇,并汇总来自多个样本的数据。我们的Loupe Browser可视化软件可协助您交互式地探索结果。
数据分析流程的输出内容
输出内容包括QC信息和文件,这些文件可使用Loupe Browser可视化软件或第三方R工具及Python工具轻松实现进一步分析。
资源
常见问题
它能够使用基于探针的方法对单个细胞的转录组(人或小鼠)进行全面分析。Feature Barcode技术的加入还能同时分析细胞表面蛋白的表达,帮助您从多个维度探索复杂生物学。此产品还支持混样分析,可提高实验的通量和效率。
一项直接比较研究的结果表明,单细胞基因表达Flex分析的灵敏度是单细胞3’基因表达分析的两倍。
样本固定可以保留细胞状态,消除使单细胞分析具有挑战性的时间限制。有了固定方案,珍贵或脆弱样本能够实现从采集点到研究中心(如中心实验室或服务商)的运输,以便进行样本处理。
固定还可以实现批量分析,简化工作流程,并降低可变性,从而提高效率。固定还能中和传染源,并且在正确验证此步骤后,可能不再需要在生物安全三级实验室(BSL-3)中处理样本。
人类和小鼠的样本都与此分析兼容。您可以从新鲜或冷冻的单细胞/单细胞核悬液或组织开始,按照我们的固定方案进行操作,也可以从FFPE组织开始,按照我们的示范方案进行操作,以便获得解离FFPE组织用于单细胞基因表达Flex分析的指引。
Fresh human and mouse samples are compatible with this option. Prior to fixation, whole cells can be labeled with antibody-oligonucleotide conjugates (using Feature Barcode technology) to profile cell surface proteins alongside gene expression.
Cell Ranger是一组分析流程,可自动生成每个细胞的表达谱,并识别具有相似表达谱的细胞簇。随后,我们的可视化软件Loupe Browser可用于交互式探索结果。这两个工具都能从我们的支持网站免费下载,您可在自己的计算机上分析10x Genomics数据集。
10x Genomics云端分析(currently available in the US and Canada only)支持您通过简单的web界面来处理单细胞基因表达Flex分析的数据,利用经过优化的可扩展架构来快速获得结果。用户可以通过这个云端界面设置和运行Cell Ranger流程。
