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ACME dissociation: a versatile cell fixation-dissociation method for single-cell transcriptomics.

ABSTRACT: Single-cell sequencing technologies are revolutionizing biology, but they are limited by the need to dissociate live samples. Here, we present ACME (ACetic-MEthanol), a dissociation approach for single-cell transcriptomics that simultaneously fixes cells. ACME-dissociated cells have high RNA integrity, can be cryopreserved multiple times, and are sortable and permeable. As a proof of principle, we provide single-cell transcriptomic data of different species, using both droplet-based and combinatorial barcoding single-cell methods. ACME uses affordable reagents, can be done in most laboratories and even in the field, and thus will accelerate our knowledge of cell types across the tree of life.

SUBMITTER: Garcia-Castro H

PROVIDER: S-EPMC8028764 | biostudies-literature | 2021 Apr

REPOSITORIES: biostudies-literature

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ACME dissociation: a versatile cell fixation-dissociation method for single-cell transcriptomics.

García-Castro Helena H Kenny Nathan J NJ Iglesias Marta M Álvarez-Campos Patricia P Mason Vincent V Elek Anamaria A Schönauer Anna A Sleight Victoria A VA Neiro Jakke J Aboobaker Aziz A Permanyer Jon J Irimia Manuel M Sebé-Pedrós Arnau A Solana Jordi J

Genome biology 20210408 1

Single-cell sequencing technologies are revolutionizing biology, but they are limited by the need to dissociate live samples. Here, we present ACME (ACetic-MEthanol), a dissociation approach for single-cell transcriptomics that simultaneously fixes cells. ACME-dissociated cells have high RNA integrity, can be cryopreserved multiple times, and are sortable and permeable. As a proof of principle, we provide single-cell transcriptomic data of different species, using both droplet-based and combinat ...[more]

PMID: 33827654

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ACME dissociation: a versatile cell fixation-dissociation method for single-cell transcriptomics

Project description:Single-cell sequencing technologies are revolutionizing biology, but are limited by the need of dissociating fresh samples that can only be fixed at later stages. We present ACME (ACetic-MEthanol) dissociation, a species-versatile cell dissociation approach that fixes cells as they are being dissociated. ACME-dissociated cells have high RNA integrity, can be cryopreserved multiple times, can be sorted by Fluorescence-Activated Cell Sorting (FACS) and are permeable, enabling combinatorial approaches of single-cell transcriptomics. ACME is based on cheap reagents and it can be done in most labs and even sampling trips. As a proof of principle, we have performed SPLiT-seq with ACME cells to obtain around ~35K cells from two planarian species and identified all previously described cell types in similar proportions. This technique allows fixed, dissociated cells to be obtained from diverse organisms that can be cryopreserved and subjected to combinatorial barcoding methods for single-cell transcriptomics and thus will accelerate our knowledge of cell types across the tree of life.

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