The development of contemporary molecular biology technology has a trend of interdisciplinary and technological interdisciplinary, Flow FISH is one of them. At present, Flow-FISH is widely used mainly as a telomere measurement technology, which combines FISH and flow cytometry. Biological telomeres have become a key cell element in aging and various diseases including cancer. Flow-FISH technology can measure the average length of telomere repeats in cells, using fluorescent peptide nucleic acid (PNA) probes that specifically bind to telomere repeats (TTAGGG) to quantify. The results obtained help to identify various diseases (telomere biology diseases, TBD) caused by genetic abnormalities in the telomerase gene. Flow FISH telomere detection has unique advantages compared with other detection schemes because it can provide results of various different cell types from one sample. In addition, Flow FISH can also analyze mRNA and protein in single cells. This is based on smFISH using flow cytometry for single-cell fluorescence measurement.
Fig 1. Comparison of mRNA expression analysis by microscopy and FISH-Flow. (Arrigucci R, et al. 2017)
Our Flow-FISH analysis service has a unique agreement for each customer. The core principle of the Telomere Flow-FISH service is to use fluorescence in situ hybridization (FISH) and labeled peptide nucleic acid (PNA) probes to determine telomere-specific repeat sequences, combined with fluorescence measurement by flow cytometry. In contrast, single-cell Flow-FISH analysis can be performed using a commercially available flow cytometer, which can provide multi-parameter information about mRNA in thousands of single cells. The process of this service includes experimental protocol customization, probe customization, flow staining, probe preparation, FISH hybridization, flow detection and data analysis.
We use FISH and labeled peptide nucleic acid (PNA) probes specific for telomere repeats combined with flow cytometry (Flow-FISH) fluorescence measurement to measure the average length of telomere repeats in cells. Flow FISH can accurately measure the median telomere length of a single cell in suspension.
Compared with other methods of measuring telomere length, Flow FISH has unique advantages and can provide multi-parameter information about the length of telomere repeats in thousands of single cells. Increased automation with the use of flow cytometers can enhance the accuracy and repeatability of measurements.
Single Cell Flow-FISH
We provide smFISH analysis services based on flow cytometry to measure mRNA expression in single cells. This method overcomes the limitations of microscope-based platforms, applies sm-FISH to flow cytometry, and uses a drain cytometer instead of a fluorescence microscope to complete data collection. Flow cytometry provides different types of information than fluorescence microscopes: microscopes provide integer counts of mRNA for each cell, while flow cytometry provides comprehensive fluorescence intensity measurements for each cell.
Microscope-based mRNA quantification is usually applied to a small number of cells attached to the glass surface and is labor-intensive. The small sample size limits the ability to assess the behavior of cell populations and precludes the identification of rare cell subpopulations that display specific expression patterns. Compared with the quantitative method using fluorescence microscopy, the advantages of flow cytometry-based measurement include a much larger number of cells analyzed, the ability to perform multi-parameter phenotype analysis, and reduced labor.
Fig 2. Flow-FISH service flow chart.
Creative Bioarray provides a comprehensive Flow-FISH measurement service to help you complete relevant analysis tests quickly and with high quality. You will benefit from our technical expertise and comprehensive experimental platform, and work with you to find the most suitable solution for your project. If you are interested in our service for measuring telomere length and mRNA quantification using Flow-FISH, please contact us for cooperation. We look forward to cooperating with you in the near future.