The Gubra approach
The molecular bench at Gubra works with everything from classic tissue specific assessments of gene expression patterns using radioactive and chromogen based ISH, to state of the art real time qPCR and RNAseq of specific cell compartments isolated by Laser Capture Microdissection.
- In situ hybridization
- qPCR analysis
- Laser capture microscopy
- HTS sequencing and bioinformatics
In situ hybridization
Revisiting an old favourite
Although it’s been replaced by qPCR in recent years, in situ hybridization is still a highly valuable tool for providing complete information about where the gene of interest is located in the tissue. In comparison to qPCR, it provides you with an estimate of total instead of relative expression. Moreover, it gives you comprehensive details on a cellular level, information that is critical to know how and where your compound impacts.
Gubra has extensive experience with P33labelled radioactive ISH (also enabling quantitative assessments) and highly sensitive chromogen/ fluorescent labelled probes for qualitative assessment of any target in any organ.
How it works
If you want to localize your target gene, a fast and often-used approach is immunostaining. This method, however, relies on the availability of specific antibodies, which often are not as specific as they need to be. A particular problem which is often ignored by the commercial suppliers of antibodies is the fact that antibodies can sometimes bind to something that isn’t your gene of interest. You'll get staining, and you think it's the gene, but it may not be. Radioactive in situ hybridization is incredibly specific, and only binds to the gene you want, giving you an accurate location of the gene of interest. As an example, we often use this method to validate our antibodies.
We use radioactively labeled RNA probes to determine the precise location of low abundance genes in various tissues and for semi-quantitative analysis of mRNA levels in different areas.
Laser capture microscopy
The Gubra approach to LCM. Gubra scientists have been working with laser capture microscopy for over 10 years. Watch our newest film to learn more about how our approach to LCM can make a difference in your study.
We use laser capture microscopy to isolate individual cells or cell groups for RNA extraction. We have perfected the sampling, cutting, RNA isolation and amplification processes, to completely avoid RNA degradation, thus allowing for gene expression analysis using qPCR, microarray or HTS sequencing.
We have more than 10 years of experience with LCM and we have perfected the sampling, cutting, RNA isolation and amplification processes, so we can profile gene expression (using qPCR, microarray or HTS sequencing) in small groups of cells in individual animals. Laser capture micro dissection enables you to fish out specific cells from heterogeneous organs and gives you a level of precision not possible with manual extraction methods.
RNAseq and Bioinformatics
We have used RNAseq in combination with LCM to demonstrate that RNAseq is not only more sensitive but also more precise than the existing microarray technologies. Don´t get frustration by information overload- advanced data analyses by skilled bioinformaticians will help you dissect and detect the target of interest.
Gubra bioinformaticians have developed a stringent data analyses platform to help you detect the target of interest from thousands of genes to a manageable number selected for further target validation All data can be combined into the rapidly expanding Gubra RNAseq database allowing you to compare genes and target across different experiments, models and organs.
Let's work together
If you have a study, and you want to know if stereology can be a useful method for gaining accurate results and true quantitative data, give us a call. By applying a methodical approach to sampling, we will help you get useful, usable results.
Contact us before you harvest
We often receive inquiries from organizations interested in using stereology, yet they have already harvested the organ, which, in many cases, impacts our ability to apply stereology in a cost- efficient manner. So if you’re interested in (or if you wish to investigate the application of stereology in your study) contact us as early in the process as possible, so we are in the best position to advise you.
of science Read
We know our results ultimately have to generate value for your company. That’s why we focus on the quality and completeness of the data we provide. We choose the methodology that will yield the most usable results for your company. Scientific research requires care, accuracy and precision. This is the delicate balance we walk. Pursuing methods with scientific merit to give you results that you can use, and that will ultimately generate value for your business.
At Gubra, we are committed to high quality deliverables. Through the strength of our quality systems and processes, from administration and study conduction to data management and reporting, we provide structured, professional and experienced services. All critical processes are governed by Standard Operating Procedures that along with quality control documentation are controlled by our Quality Assurance Group. Consistent training and evaluation processes throughout the organization ensure Gubras dynamic and evolving Quality Assurance System.
keep it simple Read
Just because it’s science, doesn’t mean the research has to be complicated. One of our mantras is the need to get clear, simple answers from research. It’s true, with limited time and resources and infinite pressure to produce results, it can be very tempting to get the most bang for your proverbial buck by trying to include as many endpoints in a study as possible. Yet, what often happens with this approach is that you get inconclusive results that require further investigation, taking more time and using more resources. By taking a straightforward approach to study design, we ensure our customers get clear answers they can use.