3 Ways FFPE Technology is Used in Cancer Research

26 Nov 2014
Kerry Parker
CEO

Editorial article

The challenge of finding high quality bio-specimens for cancer research is a huge issue, and there is limited availability of fresh tissue samples from patients in the quantities required for cellular research. The analysis of the protein pathways and cellular interactions in a tumor cell, as opposed to those of a normal cell, are essential to the understanding of the make-up of each cancer variant.

Recently, researchers have moved towards using formalin-fixed paraffin-embedded (FFPE) tissues in their research, as there are greater quantities of these available in tissue archives. However, there are many issues with the extraction of the cells, DNA and RNA from these fixed tissues. Here we summarize three different technologies that enable the use of FFPE tissues in cancer research.

Immunofluorescence imaging
In a recent study, the ability of PerkinElmer’s Vectra® multispectral imaging system and inForm data analysis software, for the analysis of control (tonsil) and clinical breast cancer FFPE samples using immunofluorescence labeling is demonstrated. Investigating signaling pathways involved in tumor progression is another important part of cancer research. Another study, using the Vectra® multispectral imaging system and inForm software, identifies the AKT, ERK, and S6 pathways in FFPE tissue sections.

RNA isolation
A recent publication in Cancer Research suggests that microRNAs (miRNAs) play a role in modulating expression of the protein special AT-rich sequence-binding protein 1 (SATB1) expression in ductual breast cancer. This protein is associated with a poor breast cancer prognosis, as it is an indicator of tumor metastasis. miRNAs were extracted from clinical FFPE samples of primary ductual breast cancer, using QIAGEN’s miRNeasy FFPE kit. Expression levels were analyzed using quantitative RT-PCR.

Flow cytometry isolation
A customer case study, based on research from the Translational Genomics Research Institute (TGen), outlines a flow cytometry-based method (using BD Biosciences BD Influx Cell Sorter) used to isolate pure populations of tumor cell nuclei solid tumor FFPE tissues, including triple negative breast carcinomas (TNBCs), glioblastomas, bladder carcinoma, and small cell carcinoma of the ovary. The group also developed a DNA extraction methodology compatible with Agilent’s Human Genome oligonucleotide array CGH kits and used whole exome sequencing analyses.

References:
doi: 10.1158/1538-7445.AM2014-5199
doi: 10.1371/journal.pone.0050586

Human Genome CGH Microarrays

Agilent Technologies

The Agilent Human Genome CGH Microarray is a dual color array containing 60-mer oligonucleotide probes synthesized in situ using Agilent’s inkjet SurePrint technology. Its comprehensive probe coverage allows genome-wide DNA copy number variation profiling while focusing on known genes, promoters, miRNAs, pseudoautosomal and telomeric regions.

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Flow Cytometry / Cell CountingFlow cytometers are used to count, sort and examine multiple characteristics of cells. Other cell analysis equipment includes image cytometers, cell counters, fluorescence-activated cell sorters (FACS), magnetic-activated cell sorters (MACS), and a range of flow cytometry assay kits. Flow cytometers can reveal information on cell viability, cell proliferation, apoptosis and cell cycle progression, as well as identify cell populations and intracellular or cell-surface molecules. Additionally, some flow cytometers, known as FACS, have an additional sorting function after analysis. Cell counters and image cytometers count live and dead cell populations and can also conduct cell proliferation assays. Find the best flow cytometers, cell counters and cell sorters in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Microarray AnalysisMicroarrays, also known as biochips, are used for the detection and analysis of multiple genes, proteins, antibodies, or biomarkers on a single microchip. This can reveal information on protein or gene expression, single nucleotide polymorphism (SNP), copy number variation (CNV), epigenetics and patient health in clinical diagnostic tests. Discover a range of microarray scanners and prefabricated antibody, protein, RNA and DNA microarrays for your analysis or consider creating your own custom microarrays with a microarray printer. Find the best microarray products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Cell-Based AssaysCell-based assays are used to monitor the presence, quantity and activities of a desired cellular analyte including drug molecules or biomarkers. This can reveal information on cell health (apoptosis, cytotoxicity, viability and proliferation assays), cell metabolism, cell migration and cell signaling mechanisms. Find the best cell-based assay products, kits and equipment with our peer reviewed product directory: compare products, check customer reviews and receiving pricing direct from manufacturers.DNA / RNA Extraction and PurificationPurified DNA and RNA are required for numerous downstream molecular biology applications. Consequently, the importance of high-quality DNA/RNA extraction and purification equipment cannot be underestimated. Many purification kits are available and are typically optimized for nucleic acid type and source, including plasmid DNA, genomic DNA, mRNA, RNA and viral nucleic acid purification kits. Automated extraction and purification of nucleic acids can be implemented with magnetic bead separator instruments or high-throughput purification workstations. Find the best DNA/RNA extraction and purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Clinical GeneticsMolecular Genetics covers the analysis of hereditary genetic disease and chromosomal abnormalities. Genetics can be analysed using DNA, RNA, and protein microarrays, PCR, RT PCR and DNA sequencing. Genetic equipment includes genetic workstations, thermal cyclers, cooling blocks and electrophoresis products. Diagnostic kits are used for DNA / RNA extraction and purification.Cancer ResearchAlthough cancer is often referred to as a single condition, it actually consists of more than 100 different diseases. Microscopy, mass spectrometry, high throughput sequencing and flow cytometry are some of the most common techniques employed in cancer research labs.CytometryCytometry is a broad term for the quantitative analysis of cells and cell systems. The most popular methods to study cytometry are flow cytometry and image cytometry.Tissue ProcessingTissue processing involves preparing biological tissue samples for microscopic analysis, including fixation, embedding, and sectioning. This process is essential for histopathology and diagnostic applications. Explore tissue processing products in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.