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ViewRNATM miRNA ISH Cell Assay for Fluorescence miRNA and RNA In Situ Hybridization (RNA FISH)

Assay Performance Highlights

ViewRNATM miRNA ISH Cell Assay

Sample Type Adherent and suspension cells
Species Any
Sensitivity Single-molecule detection
Plex Level 1 miRNA and up to 2 mRNAs
Assay Format 24-well plates*
96-well optical bottom imaging plate
Detection Fluorescent
Instrumentation Fluorescence microscope or imaging system

* 24-well plates - cover slips placed on the bottom of the 24-well plate


Simultaneous detection of miRNA, mRNA and/or protein in dissociated mouse hippocampal cultures

MAP2 and GFAP markers

Dissociated mouse hippocampal culture was stained by immunofluorescence (IF) for Map2 (neuronal cells) and Gfap (glia cells) markers to positively identify neurons and glial cells in the mixed culture (Figure 1). The same mixed culture was subjected to miRNA in situ hybridization using the ViewRNA miRNA ISH Cell Assay and probed against a neuronal specific miRNA target. Read More ...

 


Specificity of the mRNA probe in the mRNA in situ hybridization assay

visualize_miRNA_mRNA

Accurately and precisely study and visualize miRNA and mRNA of various expression levels. The assay has single-molecule RNA sensitivity – where one punctated dot corresponds to one molecule of miRNA or mRNA target. Read More ...


Monitor siRNA knockdown using the miRNA in situ hybridization assay

siRNA knockdown using miRNA in situ hybridization

Simplify gene expression studies by visualizing siRNA transfection and gene expression in the same sample. The miRNA in situ hybridization assay allows the monitoring of siRNA and the mRNA of interest for fast and easy analysis of siRNA knockdown efficiency. Read More ...

 


Visualization and quantitation of the miRNA/mRNA in situ hybridization assay

visualization and quantitation of the miRNA in situ hybridization assay

The miRNA in situ assay results can be analyzed using two analysis software: MetaMorph and CyteSeer. In this example the expression levels of four miRNA targets of various expression levels, were analyzed in HeLa cells. The results were imaged using Olympus fluorescence microscope, visualized using MetaMorh (Figure 4A) and quantified using CyteSeer (Figure 4B). Read More ...

 


miRNA/mRNA in situ hybridization assay concordance with QuantiGene 2.0 miRNA Assay

Assay Concordance

The ViewRNA miRNA ISH Cell Assay can be used to validate QuantiGene 2.0 miRNA Assay results. To evaluate concordance between these two assays, the expression levels of 10 different miRNA targets in HeLa cell was quantified using the QuantiGene 2.0 miRNA Assay. Separately, in situ miRNA analysis of the same 10 miRNA targets was performed using the ViewRNA miRNA Assay and the mean of total signal intensity for each miRNA target was quantified using CyteSeer software. Read More ...