Slide-to-Slide Arrays for High-Throughput Molecular Profiling or Rare Tumor Specimens
Stephanie E Weissinger, Peter Moller, Jochen K Lennerz. Ulm University, Ulm, Germany
Background: Tissue microarrays (TMA) have become an important tool in high-throughput molecular profiling of tissue samples in the translational research setting. Unfortunately, high-throughput profiling in small biopsy specimens or rare tumor samples (e.g. orphan diseases) is often precluded due to limited amount of tissue. Here, we present a method that allows construction of TMAs from individual 2-5 µm sections for subsequent molecular profiling.
Design: The slide-on-slide TMA technique requires a series of chemical exposures (xylene-methacrylate exchange) in combination with re-hydrated lifting, microdissection of donor-tissues into multiple small tissue fragments and subsequent re-mounting on separate recipient slides. We assessed efficacy of microdissection and mounting by number of dropouts in re-localized tissues. Quality of downstream analyses was compared with traditional assessment for the following variables: antigen-retrieval, primary antibodies, detection systems, histochemical stains, FISH as well as DNA-extractions.
Results: From a technical perspective, building a slide-to-slide array is quick reliable and cost-effective. The three most important aspects for successful re-localization of tissue sections onto the recipient array are: appropriate case selection, precise microdissection, and exact array mounting. We tested 9 different antigen retrieval techniques in combination with over 50 different antibodies, 6 histochemical stains, 8 different FISH probe-sets and several PCR-based techniques including sequencing, which all functioned appropriately. The number of drop outs ranged from 0.8-6.2%; however, we applied the same slide-on-slide technique successfully to fill these dropouts. H&E assessment of donor slides confirmed a transfer efficacy of over 93%, depending on the size of the tissue (range 76-100%). The presented technique requires manual dexterity; however, when directly compared to skills and investments in equipment and personnel for a functioning TMA facility, the tools and effort to learn and perform the slide-on-slide TMA-technique is trivial.
Conclusions: Here, we present a quick, reliable, and cost-effective method that offers the key advantages of traditional tissue microarrays – even when tissue is sparse. The perspectives of this technology in biomedical sciences are promising given that it allows researchers to create more data with less tissue.
Wednesday, March 6, 2013 9:30 AM
Poster Session V # 274, Wednesday Morning