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TotalSeq™ PhenoCyte

 

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Introducing BioLegend’s end-to-end solution for high-throughput, high-parameter single-cell protein analysis. Combining the power of our pre-titrated, optimized TotalSeq™-A antibody panels with Scale Biosciences’ Single-Cell Quantum Barcoding technology, TotalSeq PhenoCyte provides a streamlined, instrument-free workflow for scalable single-cell immunoprofiling. Blending our expertise in flow cytometry and single-cell proteomics, TotalSeq PhenoCyte can help you amplify both the scale and scope of your single-cell research.

 

Discover the technology and data below.

 

Technology Benefits

Technology Overview

 

 

BioLegend’s TotalSeq-A oligo-conjugated antibodies and cocktails enable single-cell protein profiling by sequencing using a poly(A) capture system. Optimized and integrated with Scale Biosciences’ Single Cell RNA Sequencing reagents, we have created a unique solution to high-throughput proteomics with our TotalSeq PhenoCyte Kit.

 

Scale Biosciences single-cell solutions are designed to empower your research with unmatched flexibility and scalability. Our collaboration combines the power of our TotalSeq antibodies and Multiomics Analysis Software with Scale Bio’s innovative Quantum Barcoding technology. Quantum Barcoding greatly expands the number of cells that can be analyzed with massively parallel barcoding technology by generating hundreds of millions of unique single-cell barcodes in a single workflow, while simultaneously reducing laboratory hands-on time, empowering your high throughput, single-cell protein analysis.

 

This seamless solution kit includes a lyophilized TotalSeq Universal Cocktail plus all the reagents needed through library amplification. Available in two convenient sizes (200K or 1.2M cells) to accommodate your desired output, this solution can help you unravel the complexities of cellular protein expression with ease and accuracy.

Overall Workflow

 

 

Sample staining and fixation: Cells stained with TotalSeq antibodies and subsequently fixed with paraformaldehyde.

Sample indexing (i7 primer): Cells are partitioned and indexed with various Sample Indexing Oligos (i7 primers) which hybridize to the antibody oligos.

Quantum Barcoding plate loading: Sample indexed (i7) cells are loaded onto the Quantum Barcoding plate that partition cells into microwells.

Bead loading: Quantum Barcoding protein beads containing the bead barcode (second index) are loaded onto the plate and hybridize directly to the poly(A) tail of the antibody oligos.

Capture and gap fill ligation: Post-hybridization, the beads now containing the hybridized sample index (i7) and bead barcodes are collected and undergo a gap-fill ligation to bridge the indexing sequences.

Indexing with i5 primer: The ligated product undergoes a final round of indexing via i5 indexing PCR and the library is amplified.

Library QC: Determine library size and quantification.

Sequencing: A final library consists of combinations of unique identifiers: the Sample Index, the antibody barcode, the bead barcode, and the i5 index. The identifiers help facilitate the sequencing of the libraries and with the identification of individual cells in the downstream analysis.

Multiomics Analysis Software: Data can be processed and analyzed with our free cloud-based Multiomics Analysis Software.

Note: A final library consists of combinations of three unique identifiers: the i7 index, the bead barcode, and the i5 index, which facilitate the identification of individual cells in the downstream analysis.

Molecular Details

 

Product Features

Complete End-to-Solution

 

Available in two convenient sizes (200K or 1.2M cells) to accommodate your desired output, this kit includes necessary reagents for cell staining and library amplification. For a detailed breakdown of the kit components, please visit the product Technical Data Sheet (TDS).

 

View TotalSeq™ PhenoCyte kits

 

Hashtag Multiplexing Capabilities

 

Our TotalSeq-A hashing reagents are compatible with the TotalSeq PhenoCyte kit, enabling you to increase sample throughput for more efficient workflows. For staining instructions and recommendations, please refer to the product User Manual. For additional support, please reach out to BioLegend Technical Service for assistance.

 

View all compatible Hashtags

 

Customizable Solutions for Enhanced Flexibility

 

Unlock greater flexibility by customizing this kit to fit your specific research needs by pairing it with the antibody cocktail of your choice. To explore how we can create a tailored solution for you, reach out to our Customs Solution Team for more additional information and assistance.

Sequencing and Data Analysis

 

Sequencing Requirements

 

Parameter

Description

Sequencing Depth

10K Reads per cell

Sequencing Type

Dual indexed, pair-end

Supported Sequencers

Illumina® NovaSeq 6000

Illumina® NovaSeq X

Read Cycles

Number of Cycles

Read 1: 45

Index 1: 16

Index 2: 8

Read 2: 15

 

Data Processing & Analysis Overview

 

Multiomics Analysis Software

 

To simplify data analysis, we offer our free, cloud-based Multiomics Analysis Software (MAS) with a module designed to seamlessly integrate with the TotalSeq PhenoCyte kit for data processing and analysis. Using FASTQ files as the primary input, MAS’s PhenoCyte analysis pipeline enables users to quickly and easily process and explore single-cell protein data to create publication-quality images without extensive bioinformatics knowledge.

 

MAS Benefits

 

  • A simple user interface for exploring single-cell proteomic data.
  • Accessible to all users irrespective of bioinformatics background.
  • Accessible through your web browser, no server access required.
  • Efficiently upload and process PhenoCyte FASTQ files for data alignment.
  • Uses flow cytometry-like gating based on TotalSeq antibody staining.
  • Creates UMAP, wtSNE, or TriMAP dimensionality reduction plots.
  • Easily share any workspace with collaborators or BioLegend Technical Services.
  • Exportable in *.h5ad format for use in other single-cell analysis programs.

 

Download MAS User Manual

MAS is a free cloud-based software. Before using the software suite, you will need to request an account. To request an account please fill in the form below. Please note an institutional email address is required. Generic email addresses such as Gmail, iCloud, Yahoo, etc. will not be accepted.

Request an Account

PhenoCyte Data with MAS

 

Visualizing protein expression in single-cell analysis using techniques like clustering, t-SNE, and UMAP offers a powerful way to elevate your research.

 

Immunophenotype Cells Using Flow-like Gating

 

MAS’s interactive interface allows for immunophenotyping cells with flow-like gating, offering a precise, intuitive approach to analyzing complex cellular populations.

 

 

Reveal Hidden Cell Populations

 

MAS’s clustering algorithms allow you to identify and distinguish unique cell subsets, even in complex and heterogeneous sampling, helping you uncover novel insights that could otherwise be missed with traditional methods.

 

Simplify Complex Data

 

MAS allows for a variety of dimensionality reduction techniques such as weighted t-SNE and UMAP, transforming high-dimensional protein data into intuitive, easy-to-read visualizations making it easier to graps relationships between cells and quickly spot patterns or anomalies in your data.

 

Streamline Data Interpretation

 

These visualization tools enable for intuitive exploration of large, complex data sets, turning millions of data points into actionable insights. Elevate your single-cell research, uncover new biological insights and take your data analysis to next level with the Multiomics Analysis Software.

 

Product Support

 

Find product documentation, protocols, and answers to commonly asked questions here.

 

For questions regarding product availability, shipping, and ordering, or information regarding pricing, discounts, bulk orders, and custom products contact us here.

 

For any technical questions related to TotalSeq PhenoCyte.

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