Unmatched specificity and sensitivity
The high affinity and discrimination of the miRCURY LNA™ microRNA Detection Probes result in sensitive detection of microRNAs. Specific in situ
detection of microRNA is possible in whole mounts, single cells, frozen samples and in formalin-fixed, paraffin-embedded tissue sections, including archived biobank samples. The optimized positioning of LNATM within the detection probes substantially reduces background staining.
For maximum sensitivity and detection of even lowly expressed targets, we recommend double labeled miRCURY LNA™ microRNA Detection Probes. To enable multiplexing, a range of haptens are available, to visualize the target microRNA with different combinations of FISH and/or chromogenic ISH.
Get optimal customized ISH Control probes
Custom miRCURY LNA™ microRNA Detection Probes are ideal for the detection of any small RNA sequences, but they also serve as valuable controls in ISH experiments. Our LNA™ experts can design custom probes in a number of different ways in order to confirm signal specificity of the in situ
hybridization reaction. As with any experiment, interpretation of the results is only as good as the controls. For in situ
hybridization there is no single best control, so it is preferable to have as many controls as possible, thus building confidence in the expression pattern.
Below please find examples of Custom miRCURY LNA™ microRNA Detection Probes, designed specifically for use as positive and negative control probes.
Ideal as Mismatch Negative Control probes
Mismatch probes are custom designed specifically to be used alongside the target probe. A mismatch probe differs by 1-3 nucleotides towards the target. Take full advantage of the LNA™ technology. Our LNA™ design experts will identify the optimal number of mismatches and the LNA™ positions, to reach a T
m value close to that of the target probe.
Using the same in situ
protocol on serial sections, one should obtain little or no stain with the mismatch negative control probe. An example of a custom mismatch negative control probe can be seen in Figure 1f from the publication Nielsen et al.
, 2011. High levels of microRNA-21 in the stroma of colorectal cancers predict short disease-free survival in stage II colon cancer patients. Clin Exp Metastasis. 28(1):27-38. PMID: 21069438
Target-specific positive Control probes
The short length of LNA™ probes enables alternative probes to be designed that target slightly different regions, shifted by a few nucleotides – even within the short sequence of a mature microRNA (Figure 2). Obtaining the same hybridization pattern with different probes targeting the same microRNA is a valuable positive control in ISH experiments. Our LNA™ experts custom design alternative probes with LNA™ positions and T
m value similar to that of the target probe.
Pre-microRNA detection probes
Another valuable positive control probe to rule out non-specific cross hybridization is a loop-directed probe, designed by our LNA™ experts to bind to the loop sequence of the precursor microRNA (pre-microRNA) (Figure 2). This probe should confirm the same hybridization pattern as the target probe that binds to the mature microRNA sequence. However, the expression level of the pre-microRNA is likely to be lower than that of the mature microRNA.
An example of target-specific positive control probes, including a loop-directed probe, can be seen in Figure 1 from the publication Rask et al.
, 2011. High expression of miR-21 in tumor stroma correlates with increased cancer cell proliferation in human breast cancer. APMIS. 119(10):663-73. PMID: 21917003
View experimental data on the miRCURY LNA™ microRNA Detection Probes for