[2094] 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.
Category: Techniques

Wednesday, March 6, 2013 9:30 AM

Poster Session V # 274, Wednesday Morning

 

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