Tissue microarrays, tread carefully
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Letter to the Editor Tissue microarrays, tread carefully Laboratory Investigation (2004) 84, 1677. doi:10.1038/labinvest.3700172 To the editor: Tissue microarrays (TMAs) have provided a significant technological advance for evaluating and validating antibodies for immuno- histochemistry as well as clinical biomarker devel- opment. As with any new method, there are always concerns that need to be addressed prior to wide- spread acceptance of the technique. In a key study, Camp et al showed that antigenicity is preserved in long-term stored blocks and that they are suitable for constructing TMAs. They also demonstrated the need for redundancy to ensure proper sampling of the tissue. 1 DiVito et al. 2 have taken this a step further by showing that prepared TMA slides need to be stored properly in order to prevent the loss of antigenicity. We have also made an observation that we feel is noteworthy in light of the increased use of TMAs. We recently completed a study (Mikhail et al, manuscript in preparation; same cohort was used previously in Harzan et al 3 ) examining PTEN expression in formalin-fixed paraffin-embedded whole section skin/melanoma specimens. Out of 150 section slides tested (dating back to the mid- 1970s), 16 did not show any labeling in the tumor or normal epidermis. We noticed that the unmelted paraffin wax in this subgroup of slides (from the early 1980s) had a distinctly different appearance than the others in the group. Under polarized light, the paraffin crystals were large and tetragonal-like. They ranged in size, from 10 to 50 mm (longest dimension at 20) and were haphazardly arranged. In contrast, the paraffin of the other samples consisted of small hexagonal-like crystals, measur- ing approximately 10 mm. They were uniformly and regularly arranged. The tetragonal-shaped paraffin wax did not melt at the regular incubation tempera- ture of 601C; but did at approximately 801C. After incubating at the higher temperature, 14 out of the 16 samples that did not label previously showed labeling in both tumor and normal epidermis. In addition, our staining protocol requires heat in- duced antigen retrieval (B991C) in 0.01 M citrate for 20 min in a 1200 W microwave oven. Interestingly, antigen retrieval did not initially ‘unmask’ the antigen in these samples; only melting at the higher temperature followed by antigen retrieval induced immunoreactivity. This observation suggests that it is still important to take into consideration the source of the tissue blocks when constructing TMAs so that false positives and negatives are minimized. This is essential if multi-institutional studies are performed where tissues may be collected from several differ- ent hospitals. We commend Camp and DeVito for their efforts in addressing important issues in the use of TMAs. Luis Chiriboga 1 , Iman Osman 2 , Maryann Mikhail 2 and Christie Lau 1 1 Department of Pathology and 2 Department of Dermatology, NYU School of Medicine & Bellevue Hospital, 462 First Avenue, New York, NY 10016, USA References 1 Camp RL, Charette LA, Rimm DL. Validation of tissue microarray technology in breast carcinoma. Lab Invest 2000;80:1943–1949. 2 DiVito KA, Charette LA, Rimm DL, et al. Long-term preservation of antigenicity on tissue microarrays. Lab Invest 2004;84:1071–1078. 3 Hazan C, Melzer K, Panageas KS, et al. Evaluation of the proliferation marker MIB-1 in the prognosis of cuta- neous malignant melanoma. Cancer 2002;95:634–640. Laboratory Investigation (2004) 84, 1677 & 2004 USCAP, Inc All rights reserved 0023-6837/04 $30.00 www.laboratoryinvestigation.org
Transcript of Tissue microarrays, tread carefully
Letter to the Editor
Tissue microarrays, tread carefully
Laboratory Investigation (2004) 84, 1677. doi:10.1038/labinvest.3700172
To the editor: Tissue microarrays (TMAs) haveprovided a significant technological advance forevaluating and validating antibodies for immuno-histochemistry as well as clinical biomarker devel-opment. As with any new method, there are alwaysconcerns that need to be addressed prior to wide-spread acceptance of the technique. In a key study,Camp et al showed that antigenicity is preserved inlong-term stored blocks and that they are suitable forconstructing TMAs. They also demonstrated theneed for redundancy to ensure proper sampling ofthe tissue.1 DiVito et al.2 have taken this a stepfurther by showing that prepared TMA slides needto be stored properly in order to prevent the loss ofantigenicity.
We have also made an observation that we feel isnoteworthy in light of the increased use of TMAs.We recently completed a study (Mikhail et al,manuscript in preparation; same cohort was usedpreviously in Harzan et al3) examining PTENexpression in formalin-fixed paraffin-embeddedwhole section skin/melanoma specimens. Out of150 section slides tested (dating back to the mid-1970s), 16 did not show any labeling in the tumor ornormal epidermis. We noticed that the unmeltedparaffin wax in this subgroup of slides (from theearly 1980s) had a distinctly different appearancethan the others in the group. Under polarized light,the paraffin crystals were large and tetragonal-like.They ranged in size, from 10 to 50mm (longestdimension at � 20) and were haphazardly arranged.In contrast, the paraffin of the other samplesconsisted of small hexagonal-like crystals, measur-ing approximately 10mm. They were uniformly andregularly arranged. The tetragonal-shaped paraffinwax did not melt at the regular incubation tempera-ture of 601C; but did at approximately 801C. Afterincubating at the higher temperature, 14 out of the16 samples that did not label previously showed
labeling in both tumor and normal epidermis. Inaddition, our staining protocol requires heat in-duced antigen retrieval (B991C) in 0.01 M citrate for20 min in a 1200 W microwave oven. Interestingly,antigen retrieval did not initially ‘unmask’ theantigen in these samples; only melting at the highertemperature followed by antigen retrieval inducedimmunoreactivity.
This observation suggests that it is still importantto take into consideration the source of the tissueblocks when constructing TMAs so that falsepositives and negatives are minimized. This isessential if multi-institutional studies are performedwhere tissues may be collected from several differ-ent hospitals. We commend Camp and DeVito fortheir efforts in addressing important issues in theuse of TMAs.
Luis Chiriboga1, Iman Osman2,Maryann Mikhail2 and Christie Lau1
1Department of Pathology and2Department of Dermatology,
NYU School of Medicine & Bellevue Hospital,462 First Avenue, New York,
NY 10016, USA
References
1 Camp RL, Charette LA, Rimm DL. Validation of tissuemicroarray technology in breast carcinoma. Lab Invest2000;80:1943–1949.
2 DiVito KA, Charette LA, Rimm DL, et al. Long-termpreservation of antigenicity on tissue microarrays. LabInvest 2004;84:1071–1078.
3 Hazan C, Melzer K, Panageas KS, et al. Evaluation of theproliferation marker MIB-1 in the prognosis of cuta-neous malignant melanoma. Cancer 2002;95:634–640.
Laboratory Investigation (2004) 84, 1677& 2004 USCAP, Inc All rights reserved 0023-6837/04 $30.00
www.laboratoryinvestigation.org
