Overview of the Applied Biosystems® Arcturus® Laser Capture Microdissection (LCM) Process
Viability of the IR laser-enabled Laser Capture Microdissection (LCM) technique has been shown fora variety of different cell types. The procedure is remarkably simple and robust and available exclusively with Applied Biosystems® Arcturus® Laser Capture Microdissection (LCM) instruments. Any Arcturus® LCM system can be used, including the new, open and modular ArcturusXT™ . The Arcturus® LCM method is extraordinarily gentle and ideal for microdissection of single cells or small numbers of cells. This system uniquely combines LCM and UV laser cutting for ultimate microdissection flexibility.
Simple four-step process to capture cells and recover biomolecules
After locating the cells of interest, a CapSure® LCM Cap is placed over the target area. Pulsing the laser through the cap causes the thermoplastic film to form a thin protrusion that bridges the gap between the cap and tissue and adheres to the target cell. Lifting of the cap removes the target cell(s) now attached to the cap. The isolated cells are now available for extraction of biomolecules.
Complete System for Microgenomics
Applied Biosystems® combines Arcturus® Laser Capture Microdissection (LCM) systems, instruments and microgenomics reagents to give you the industry’s only complete solution for your microgenomic research requirements.
To identify target cells for laser capture microdissection, tissue preparation kits stain tissues to aid in identification and sourcing of biomolecules for further analysis.
HistoGene® Stain is a special solution developed by Arcturus® to stain tissues for LCM subsequently used as sources of RNA, DNA and proteins. It is a fast penetrating stain that provides good contrast by differential staining of nuclei (purple) and cytoplasm (light pink). By minimizing the exposure of tissues to water where nucleases may be activated, the HistoGene® staining process helps to preserve RNA integrity that may be otherwise compromised when using longer staining protocols.
Identify target cells for LCM and gene expression analysis using antigen markers to highlight specific surface or intracellular proteins. The HistoGene® LCM Immunofluorescence Staining Kit is the only kit specifically designed for retrieval of high-quality RNA from immunofluorescently stained frozen tissue.
Paradise® Stain is a special solution developed by Arcturus® to stain formalin-fixed paraffin-embedded (FFPE) tissues for LCM subsequently to be used as sources of RNA, DNA and proteins. The fast penetrating stain provides good contrast by differential staining of nuclei (purple) and cytoplasm (light pink). By minimizing the exposure of tissues to water where nucleases may be activated, the Paradise® staining process helps to preserve RNA integrity that may be otherwise compromised when using longer staining protocols.
Step 2: Select Cells
Once the tissue is stained, the cells of interest are located and selected using specialized microdissection software. The IR laser activates a transfer film on a cap placed on the tissue sample, adhering the film to the underlying cells of choice. The cells of interest are then lifted away from the tissue section, leaving all unwanted cells behind and providing a pure population for cell-specific molecular analysis.
The ArcturusXT™ system combines infrared (IR) Laser Capture Microdissection (LCM) infrared (IR) and Ultraviolet (UV) Laser Cutting in one platform. The open, modular design of the ArcturusXT™ system provides unparalleled research flexibility and versatility. The system removes the guesswork from the microdissection process by allowing researchers to maintain custody of the sample throughout the experiment, ensuring that only the desired material has been collected.
CapSure® LCM Caps enable the precise and rapid extraction of populations of pure cells from tissue sections and cytology samples with Arcturus® Laser Capture Microdissection.
Step 3: Purify DNA/RNA
DNA or RNA is purified from the sample for downstream analysis.
The PicoPure® DNA Extraction Kit offers an easy, streamlined genomic extraction procedure that produces PCR-ready DNA. Extract and amplify DNA in the same tube, without organic extraction or spin columns. Recover DNA from as few as ten cells procured by Laser Capture Microdissection (LCM) or from milligrams of tissue.
The PicoPure® RNA Isolation Kit is designed to recover high-quality total RNA consistently from fewer than ten cells enabling use with small samples containing picograms of RNA and samples containing up to 100 µg of RNA. Since the RNA elutes in as little as 10 µl of buffer, it is ready for gene expression analysis without any vacuum concentration.
The Arcturus® Paradise® PLUS Reagent System enables unprecedented gene expression analysis of millions of previously inaccessible samples. This integrated solution for gene expression studies is optimized for using sample from formalin-fixed, paraffin-embedded (FFPE) tissues, and includes reagents for tissue staining, RNA isolation and RNA amplification. Amplify RNA from as little as 5 ng of fixed RNA using methods that have been specially optimized to overcome the challenges of cross-linked templates.
The RiboAmp® PLUS RNA Amplification Kit offers unique linear amplification from as little as 1 ng of total RNA. Using a proprietary, patent-pending process for high-efficiency, high-fidelity linear amplification, messenger RNA is amplified up to 1000-fold in one synthesis round and up to 1,000,000-fold in two rounds. The RiboAmp® PLUS Kit produces amplified antisense RNA (aRNA), ready for labeling and microarray hybridization or QRT-PCR.
The RiboAmp® PLUS RNA Amplification Kit enables you to perform molecular analysis on RNA samples that, until now, have been too small for microarray analysis. This Kit offers unique linear amplification from as little as 1 ng of total RNA.
The Arcturus® Paradise® PLUS Reagent System enables unprecedented gene expression analysis of millions of previously inaccessible samples. This integrated solution for gene expression studies is optimized for using sample from formalin-fixed, paraffin-embedded (FFPE) tissues, and includes reagents for tissue staining, RNA isolation and RNA amplification. Amplify RNA from as little as 5 ng of fixed RNA using methods that have been specially optimized to overcome the challenges of cross-linked templates.
Step 5: Label RNA
The amplified RNA (aRNA) is labeled for gene expression profiling and analysis to represent the mRNA transcript for downstream analysis.
Turbo Labeling™ Kits offer a proprietary, non-enzymatic technology optimized for the labeling of unmodified, amplified RNA (aRNA) for gene expression profiling. The unmodified aRNA is labeled post-amplification, resulting in unmodified aRNA with higher yields and longer aRNA fragments, to allow for better representation of the mRNA transcript for downstream analysis.
For research use only. Not intended for any animal or human therapeutic or diagnostic use.
