CellCover was developed for fast “one step” stabilizing of biomolecules in life science research. CellCover is a non-toxic formulation for protecting status of expression in human and animal cells, solid tissues, including tumors and cultured cells (adherent, suspension, spheroids). DNA, RNA and proteins are all protected.
- Stabilize RNA and proteins simultaneously for several days
- Cells maintain close to in vivo morphology without chemical crosslinking of biomolecules allowing straight forward RNA isolation.
- No freezing necessary
How it works
As soon as cells get exposed to CellCover their metabolic state is frozen without applying low temperatures- liquid frozen. 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, both leading to exceptional stabilization of DNA, RNA and protein. This is beneficial e. g. for analysis by flow cytometry or in next generation or RNA sequencing approaches.
Cells grown on almost any substrate could be treated with CellCover (liquid freezing) to maintain in vivo morphology without chemical crosslinking of biomolecules during cell fixation. RNA degradation is inhibited leading to great results in RNA sequencing or single cell sequencing experiments. Material destined for a biobank is kept without altering expression due to environmentally induced gene expression changes.
Protein expression analysis has no limits. Compared to conventional formaldehyde fixation, formalin sensitive epitopes are readily accessible for antibodies in immunohistochemical experiments.
High RIN Values
RNA degradation is prevented: Even integrity of high molecular weight RNA is protected (e.g precurser rRNA, which are normally degraded after applying commonly used standards procedures) – and there is no need to hurry! You can store your sample in CellCover at 4°C and isolate RNA later.
1.1: Standard 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 analysed with standard procedures. Results show a stable RIN value of 10 from first till last day of the time course.
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.
In comparison to commonly used molecular biological methods, CellCover adds following new analytical tools to bio-molecular research:
- Stabilize RNA and Protein simultaneously for several days, without doubt of molecule integrity
- 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.