CellCover is the only reagent that allows parallel storage of proteins, RNA, and DNA in their cellular context.
- Maintain cellular shape integrity without chemical crosslinking.
- Protect DNA, RNA and proteins, protein expression and gene expression status.
- Instant stabilization in one fast step.
- Compatible with staining (in CellCover).
- Non-toxic, ready to use solution.
- For use in human and animal cells and solid tissues, including tumors and cultured cells (adherent, suspension, spheroids).
- Truly representative molecules for genomic analysis, transcriptome analysis, and proteomic analysis.
- Virtually all downstream applications are compatible with cell fixation using CellCover, including single cell RNA sequencing, NGS, protein sequencing, and more.
Cells Treated with CellCover Maintain Their Native State.
Please scroll down for more information on how to use CellCover.
How to use CellCover
CellCover stabilizes cells in suspension, biopsies, or tissue without cell lysis, keeping cells close to in vivo morphology. CellCover is compatible with morphological analyses and staining procedures, including immunocytochemistry, immunohistochemistry, and flow cytometry, among many other downstream applications.
A unique and special feature of CellCover is the ability to perform additional molecular analysis on cells that have already been analyzed on a morphological or molecular level: DNA (e.g. PCR), RNA (e.g. RNA seq or single cell RNA-seq) or protein (e.g. protein modification analysis, protein sequencing, or proteomic analysis) can be isolated for subsequent applications after initial immunolabeling, immunohistochemistry, immunocytochemistry, or flow cytometry analyses.
Application of CellCover is simple. Remove cell culture medium, apply CellCover and incubate for two minutes. Then proceed with your standard protocol.
CellCover can be used in the following applications (and many other!):
- Batch and single cell analysis
- Flow cytometry / FACS
- DNA Sequencing
- Protein Sequencing
- RNA Sequencing (RNA Seq)
- Single Cell RNA Sequencing
- Northern Blotting
- Western Blotting
- Many more applications
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How it works
CellCover was developed for fast “one step” stabilization of biomolecules in life science research. CellCover is a non-toxic formulation, a formaldehyde alternative designed to protect the status of DNA, RNA, and protein expression and gene expression in human and animal cells and solid tissues, including tumors and cultured cells (adherent, suspension, spheroids). DNA, RNA, proteins and cell morphology are all protected.
As soon as cells are exposed to CellCover their metabolic state is frozen without applying low temperatures- the liquid frozen effect. Synthesis and turnover of bio-molecules are stalled. This includes instant disruption of cellular degradation pathways, making cell fixation by CellCover very fast. Chemical degradation is inhibited with CellCover as well, leading to exceptional protein stabilization and protein fixation, as well as RNA fixation and DNA fixation. CellCover acts as an DNA and RNA stabilizer and shield, which is beneficial e. g. for analysis by flow cytometry or in NGS or RNA seq approaches. CellCover is a key initial step for successful experiments in genome analysis, transcriptome analysis, and proteomic analysis.
Cells grown on almost any substrate can be treated with CellCover by liquid freezing to maintain in vivo morphology without chemical crosslinking of biomolecules during cell fixation.
Cell morphology is preserved, allowing for accurate visualization of molecule location. DNA, RNA, and proteins are also stabilized and preserved when performing cell fixation with CellCover, allowing for effective downstream experiments in genomics, transcriptome analysis, and proteomics.
High RIN Values
With CellCover performing RNA fixation and acting as an RNA stabilizer, RNA degradation is inhibited, leading to great results in RNA sequencing or single cell RNA seq or single cell sequencing experiments. Material destined for a biobank is kept without altering expression due to environmentally induced gene expression changes. Even integrity of high molecular weight RNA is protected (e.g. precursor rRNA, which are normally degraded after applying commonly used standard procedures) – and there is no need to hurry! You can store your sample in CellCover at 4°C and isolate RNA later.
Protein expression analysis has no limits. Study protein modification and proteomics with CellCover taking care of protein fixation and protein stabilization. CellCover is a safe formaldehyde alternative fixation, with formalin sensitive epitopes readily accessible for antibodies in immunohistochemistry experiments.
1.1: Standard RNA Integrity Number (RIN) Analysis of RNA
1.2: Cultured cells (SKMel-28) stored for up to 12 days in CellCover. RNA was isolated and its integrity was analyzed with standard procedures. Results show a stable RIN value of 10 from first till last day of the time course. High RIN is important for many gene expression and transcriptome analysis experiments, including batch RNA seq or single cell RNA-seq / single cell RNA seq.
A unique and special feature of CellCover liquid freezing is that you can perform additional molecular analysis on cells already analysed on morphological or molecular level: DNA (for e.g. PCR), RNA (for e.g RNA seq) or protein (for e.g protein modification analysis) can be isolated for subsequent applications after initial IHC, ICC or flow cytometry analyses.
These unique properties now enable new types of analyses of cellular functions. With CellCover liquid freezing, subpopulations of cells can be separated according to known properties by flow cytometry and subsequently analysed for gene expression profiles on RNA or on protein-level. Thus, with CellCover liquid freezing it is easy to link transcriptome and proteome of a given cell population.
(Left: CK Pan, Right: CD31)
CellCover stabilizes cells in suspension, biopsies or tissue. CellCover is compatible with morphological analyses and staining procedures, with immunocytochemistry and flow cytometry.
(Initial protein analysis & RNA isolation)
Cells treated with CellCover maintain morphology for several days. Epitopes are protected and can be stained later. Highly intact RNA can be isolated from cells in which protein has already been stained by immunolabeling. Quality of RNA after CellCover protection: Cells have been immunostained for protein, RNA was isolated subsequently.
(Left: Formalin, Right: CellCover)
CellCover liquid freeze effect allows visualization of formalin sensitive epitopes like Vimentin, enabling new insights in cells structure and architecture.
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In comparison to commonly used molecular biological methods, CellCover adds following new analytical tools to bio-molecular research:
- Stabilize RNA and proteins simultaneously for several days, without doubt of molecule integrity
- Cells maintain close to in vivo morphology without chemical crosslinking of biomolecules allowing straight forward RNA isolation.
- No freezing necessary
- Reliable analysis HMW-RNA and low stability enzymes even after day of storage
- Apply immunohistochemistry on formalin sensitive epitopes
- Apply in situ hybridization for analysis of gene expression
- Store cells conveniently for several days without change of protein or RNA expression profile or change in cells morphology as found with methanol
- Isolate subpopulations by flow cytometry based on protein labelling and perform subsequent transcriptome and/or proteome (MALDI) analyses.
- Next generation sequencing and single cell RNA seq.
- Reliable sample collection for biobank deposits.