In the intricate domain of molecular biology, the advent of advanced techniques has made provision for scientists to hunt through deeper into the inner workings of cells and untangle the secrets hidden within. One such ground breaking technology that has emerged is sequential fluorescence in situ hybridization is a powerful molecular imaging technique used in the field of biology and transcriptomics to study the spatial organization and interactions of RNA molecules within individual cells and tissues. It is a type of RNA fluorescence in situ hybridization (RNA-FISH) that enables researchers to visualize the locations of multiple RNA species simultaneously with single-molecule sensitivity.
This blog post takes you on a journey through the fascinating world of sequential RNA-FISH (Omicsveu’s RNAveu) shedding light on its significance, applications, and potential to revolutionize our understanding of cellular processes.
Unveiling the Invisible: What is RNAveu?
RNAveu is a cutting-edge molecular imaging technique that allows scientists to peer into the heart of cells and tissues like never before. By combining the power of fluorescence microscopy and hybridization techniques, OmicsVeu’s RNAveu technology enable researchers to visualize and analyze multiple RNA molecules within the context of individual cells, tissues, and even whole organisms with exceptional precision. Unlike traditional techniques that primarily focus on protein expression, OmicsVeu’s sequential RNAveu offers a window into the world of gene expression by targeting specific RNA sequences (Figure 1).
Figure 1: Expression of PD1(ornage), PDL1 (Green), TIM3 (Yellow) and LAG3(Red) in Non-small cell lung carcinoma (nucleus- Blue)
How does RNAveu Work?
RNAveu process starts with series of innovative steps that combine principles of in situ hybridization and signal amplification. Here’s a simplified breakdown of the process:
- Probe Design: multiple short oligonucleotide probes, typically 25-30 bases long, are designed to be complementary to the target mRNA sequence. These probes are conjugated with unique barcode oligonucleotides typically 30-32 bases long.
- Complimentary barcode probe: A short oligonucleotide probes (25 bases long) are designed to be complimentary of unique barcode sequence. These probes are tagged with five fluorescent molecules each side of the oligo strand.
- Tissue Preparation: Tissues or cells of interest are carefully preserved and sectioned to maintain their structural integrity.
- Hybridization: The labeled RNA probes are applied to the tissue sections. These probes hybridize (bind) specifically to the target RNA sequences within the cells.
- Signal Amplification: Complimentary probe hybridization steps are performed to enhance the signal generated by the bound probes. This greatly increases the sensitivity of detection.
- Visualization: The labelled probes, now amplified, produce a visible signal that can be detected using fluorescence microscopy.
Multiple cycles of Revelation
OmicsVeu’s RNAveu builds colorful tissue morphology of RNA expression. It does this by performing multiple rounds of hybridization and imaging.
Here’s how it works:
- A set of probes specific to a particular RNA molecule is applied to the slide.
- The slide is imaged to capture the glowing spots where the probes have bound.
- The fluorescent markers are chemically removed or bleached, resetting the same slide for the next round.
- Steps 1 to 3 are repeated, each time using a new set of probes for different RNA molecules.
- The cumulative result is a detailed snapshot of the spatial distribution of various RNA molecules within a cell or tumor microenvironment.
Applications of RNAveu:
OmicsVeu’s RNAveu has proven to be a versatile tool with a wide range of applications across various fields of research:
- Cancer Research: Understanding the gene expression patterns in cancer cells can provide insights into disease progression and potential therapeutic targets.
- Neuroscience: It allows researchers to visualize gene expression in specific regions of the brain, aiding in the study of neurological disorders and brain development.
- Infectious Diseases: RNAveu has been instrumental in studying viral infections by enabling the visualization of viral RNA within infected cells.
- Pharmacology: Assessing the effects of drugs on gene expression patterns provides valuable information for drug development.
Advantages of RNAveu:
RNAveu offers several advantages over traditional techniques. It provides single-cell resolution, allowing researchers to analyze heterogeneity within a tissue sample. Additionally, it enables spatial analysis, offering insights into the localization of mRNA molecules within cells.
Looking ahead, this sequential RNA-FISH holds the potential to reshape our understanding of cellular processes and disease mechanisms. As the technique continues to evolve, it’s likely to become an even more indispensable tool in molecular and cellular biology research.
Sequential RNA-FISH (RNAveu) is a trailblazer in the realm of spatial transcriptomics, a field dedicated to understanding how RNA are expressed in their native context within tissues. It’s opening doors to even more advanced techniques that can simultaneously analyze thousands of genes across entire tissue sections, providing a comprehensive view of gene expression landscapes. Its ability to uncover the cellular profiling of mRNA within cells is transforming our understanding of tumor biology, development, and diseases. As RNAveu continues to evolve and inspire new technologies, the invisible world of cells is becoming increasingly visible, addressing a more accurate and awe-inspiring idea of personalized medicine.