A single round of amplification yields product sizes ranging from 200 bases to 6 kb. The majority of these products are approximately 1.5 kb in length. A second round of amplification will result in shorter products. We recommend using an Agilent 2100 bioanalyzer to visualize these products. Amplification products can be visualized by agarose gel electrophoresis; they will migrate as a smear. Although this data is still useful, it is less informative than bioanalyzer analysis.
Glass microarray analysis experiments typically require 5-20 µg of total RNA per slide for sample labeling and hybridization. Thus, microarray-based gene expression analysis of very small samples [laser capture microdissection (LCM), tissue biopsies, or other clinical samples] is difficult due to the very low amounts of total RNA recovered from the samples. Linear amplification of RNA from small samples produces sufficient quantities of RNA for sample labeling and hybridization. Since the amplification technique is highly reproducible and maintains representation of the gene expression in the original sample, it is recommended for probe synthesis by most manufacturers of commercially available microarrays.
How do I locate the overview image on the Arcturus XT LCM Instrument?
First, optimize the visualization of the image in the live video at 2X by increasing or decreasing the brightness setting, then right-click on the overview image and select "remember settings". This resets your optimized image settings, and when you select "re-acquire overview image" you will now generate a perfectly exposed overview image.
With the ArcturusXT LCM Instrument, which laser should I use to isolate my region of interest? UV cutting laser or IR capture laser?
For a few cells: The gentle IR-only approach is the best recommendation for preserving nucleic acids and verifying that the desired material is collected on the cap.
For groups of cells or large tumor regions: Mount your tissue on a membrane slide and take advantage of the power of the IR, or IR + UV cutting laser to cut around your region of interest quickly and cleanly.
What is Arcturus Laser Capture Microdissection (LCM)?
The ArcturusXT LCM Instrument has a proprietary combination of a gentle IR laser and a powerful UV laser that work in conjunction to efficiently isolate cells from frozen sections or paraffin embedded sections without changing morphology or integrity of the biological content. The IR laser helps to capture the cells of interest, and the UV laser microdissects the captured cells. This is done without affecting the morphology of the cells, and allows for visual inspection of the remaining specimen to help ensure the quality of the sample collected. In addition to the flexibility that the dual laser system provides, there is the flexibility of multiple stage inserts for various sample types. A wide-slide stage format is available for neurobiology researchers working with very large sections of brain tissue. The petri dish stage insert enables live-cell microdissection applications such as stem cell research and other rare-cell isolations.