in situ HYBRIDIZATION : VISUALIZATION AND QUANTIFICATION OF THE GENE EXPRESSION

in situ hybridization is a technique for visualizing specific RNA or DNA within cell or tissue. Its principle is based on the use of a fluorescent probe which represents a nucleic acid sequence specific to a target-gene.

This technology offers quantitative and visual information about the studied mRNA. It is an alternative to immunohistochemistry or immunofluorescence experiments when there is no available antibody. in situ hybridization can also be of interest for the analysis of  specific gene expression profiles in sensitive samples, where total RNA extraction by RT-qPCR may  not provide satisfactory results.

Bioalternatives can assist you in the implementation and in the analysis of your in situ hybridization experiments.

Characteristics:

  • Good sensitivity (detection of one mRNA  molecule per cell)
  • Good specificity
  • Low noise
  • 3000 available probes and  possibility of designing customized probes
  • Up to 3 different probes in one cell
  • Can be paired with immunohistofluorescence experiments

Applications:

  • Visualization of gene expression
  • Visualization of co-expression of miRNAs and of their target-genes
  • Visualization of the kinetics of gene expression profiles
  • Quantification of the expression of genes of interest
  • Validation of biomarkers in a given cellular typ
  • Validation of knock-out experiments

The Panomics QuantiGene® technology used in Bioalternatives can analyse up to 3 different probes in the same cell. The principle of this technology is based on a fluorescent signal amplification method, by using bDNA(branched DNA) technology.

Data acquisition in in situ hybridization experiments is performed using a fluorescence microscope or with a cell imaging system, such as INCELL Analyzer.

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