Although RISH has been around for over 40 years, most of the current RISH protocols produce high background signals due to poor probe design and improper hybridization and washing procedure. These protocols usually use long probes (over 40nt) and relatively low temperature (20C below annealing temperature) for hybridization and washing. Long probes can be easily trapped on cell matrix and at low washing temperature non–specifically bound probes cannot be completely removed after washing, resulting in a high background RISH image. OrioR Lab has established a probe design algorithm that uses only 15-20 nt target-specific sequence for probe synthesis to eliminate non-specific probe hybridization. For each probe highest hybridization and washing temperature will be tested to remove non-specific binding of the probes to eliminate background signal. Precise temperature control is achieved by using micro chip controlled equipments for hybridization and washing. This accurate RNA in situ hybridization ( AccuRISH) technology developed by OrioR Lab, LLC produces the most accurate RNA detection image with no cross binding of probe and generates very low background signal. With AccuRISH , for the first time, precision localization of any RNA targets at subcellular level can be achieved.
In Situ Hybridization Probe Design and Labeling
Our bioinformatics specialist will work with you to design probe for your application. We work with you to choose ideal probe length, location and labeling chemistry to ensure best in situ hybridization results.
CISH and FISH service
Our service laboratory can perform both Chromagenic In Situ Hybridization (CISH) or Fluorescence In Situ Hybridization (FISH) depending on our customer's application and requirements. We can perform CISH or FISH on both tissue sections or prepared cell slides.
Tissue processing: fixed tissue samples are processed and embedded into a paraffin block for subsequent sectioning and staining. The slides can be coated or uncoated according to customer's specification.
H&E Stain - The standard histology stain that differentiates the nucleus from the cytoplasm.
Unstained Sections - Unstained sections can be produced for further processing by the customer.
Serial Sections - Consecutive sections are taken from a tissue block and placed in order on a set of slides.
Step Sections - If you wish to scan a tissue without looking at every section, sections can be taken at designated intervals as the tissue is sectioned.
RNAse free slide preparation - Slides are prepared with RNAse rfree precautions to minimize loss of RNA by RNAses. Blocks are sectioned with new blades and placed in DEPC-H2O and mounted onto slide. Gloves are worn to prevent cross-contamination.
Targeted sectioning using a microscope - If you want to locate a particular area of interest, our experienced histotechnologists can expertly locate the area by mounting unstained sections and confirming the area with a microscope.
Sections placed in tubes - Tissues can be sectioned and individually placed in microfuge tubes for subsequent procedures (eg. PCR).
IHC and IF
We provide customized immunohistochemistry, immunofluorescence and nucleic acid detection services:
In depth consultations to assess staining strategies. Includes customized antibody and control tissue searches -- many human control tissues and cell lines are already available
Chromogenic or fluorescent detection methods offered
Multiplex staining available