ArticlePloS one2021
Glyoxal fixation facilitates transcriptome analysis after antigen staining and cell sorting by flow cytometry.
Article in PloS one, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
18 citing papers in PubMed, 36 citations in OpenAlex.
- Could glyoxal be an alternative fixative to paraformaldehyde in human spermatozoa?Journal of assisted reproduction and genetics · 2026Article
- Single-nucleus chromatin accessibility and gene expression co-profiling by ISSAAC-seq.Nature protocols · 2026Review
- An at-home blood collection device for remote immune monitoring by high-parameter flow cytometry.JCI insight · 2026Article
- Phenotypic Characterization and Sorting of Circulating Tumor Cells (CTCs) Using a Digital Image-Based Sorter.Methods in molecular biology (Clifton, N.J.) · 2026Article
- The cis-regulatory logic integrating spatial and temporal patterning in the vertebrate neural tube.Developmental cell · 2025Article
- Functional phenotyping of genomic variants using joint multiomic single-cell DNA-RNA sequencing.Nature methods · 2025Article
- Evolutionarily recent transcription factors partake in human cell cycle regulation.Cell genomics · 2025Article
- Glyoxal is a superior fixative to formaldehyde in promoting antigenicity and structural integrity in murine cardiac tissues.Journal of molecular and cellular cardiology plus · 2025Article
- DEPICT-seq: Single-Cell Transcriptomic Analysis of Rare Cell Subsets Isolated via Nucleic Acid Cytometry.Analytical chemistry · 2024Article
- Streamlined Full-Length Total RNA Sequencing of Paraformaldehyde-Fixed Brain Tissues.International journal of molecular sciences · 2024Article
- Defining an Optimized Workflow for Enriching and Analyzing Residual Tumor Populations Using Intracellular Markers.The Journal of molecular diagnostics : JMD · 2024Article
- Single-Cell Transcriptome Analysis of Acute Myeloid Leukemia Cells Using Methanol Fixation and Cryopreservation.Diseases (Basel, Switzerland) · 2023Article
- Glyoxal as an alternative fixative for single-cell RNA sequencing.G3 (Bethesda, Md.) · 2023Article
- Evolutionary trade-offs constraining the MHC gene expansion: beyond simple TCR depletion model.Frontiers in immunology · 2023Article
- Escape from G1 arrest during acute MEK inhibition drives the acquisition of drug resistance.NAR cancer · 2022Article
- Glyoxal Fixation Is Optimal for Immunostaining of Brain Vessels, Pericytes and Blood-Brain Barrier Proteins.International journal of molecular sciences · 2022Article
- From phenotypical investigation to RNA-sequencing for gene expression analysis: A workflow for single and pooled rare cells.Frontiers in genetics · 2022Article
- Dissecting viral infections, one cell at a time, by single-cell technologies.Microbes and infectionReview
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
A simple method for extraction of high quality RNA from cells that have been fixed, stained and sorted by flow cytometry would allow routine transcriptome analysis of highly purified cell populations and single cells. However, formaldehyde fixation impairs RNA extraction and inhibits RNA amplification. Here we show that good quality RNA can be readily extracted from stained and sorted mammalian cells if formaldehyde is replaced by glyoxal-a well-characterised fixative that is widely compatible with immunofluorescent staining methods. Although both formaldehyde and glyoxal efficiently form protein-protein crosslinks, glyoxal does not crosslink RNA to proteins nor form stable RNA adducts, ensuring that RNA remains accessible and amenable to enzymatic manipulation after glyoxal fixation. We find that RNA integrity is maintained through glyoxal fixation, permeabilisation with methanol or saponin, indirect immunofluorescent staining and flow sorting. RNA can then be extracted by standard methods and processed into RNA-seq libraries using commercial kits; mRNA abundances measured by poly(A)+ RNA-seq correlate well between freshly harvested cells and fixed, stained and sorted cells. We validate the applicability of this approach to flow cytometry by staining MCF-7 cells for the intracellular G2/M-specific antigen cyclin B1 (CCNB1), and show strong enrichment for G2/M-phase cells based on transcriptomic data. Switching to glyoxal fixation with RNA-compatible staining methods requires only minor adjustments of most existing staining and sorting protocols, and should facilitate routine transcriptomic analysis of sorted cells.
Indexed as
Identifiers
What OpenQuestion holds
Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.