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LETTER TO EDITOR  
Year : 2011  |  Volume : 54  |  Issue : 3  |  Page : 654-655
Construction of tissue arrays without prefabricated recipient paraffin block experience of a novel technique in resource poor settings


Department of Pathology, Shrimati Kashibai Navale Medical College and General Hospital, Pune, India

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Date of Web Publication20-Sep-2011
 

How to cite this article:
Pathak GS, Deshmukh SD, Ashturkar AV. Construction of tissue arrays without prefabricated recipient paraffin block experience of a novel technique in resource poor settings. Indian J Pathol Microbiol 2011;54:654-5

How to cite this URL:
Pathak GS, Deshmukh SD, Ashturkar AV. Construction of tissue arrays without prefabricated recipient paraffin block experience of a novel technique in resource poor settings. Indian J Pathol Microbiol [serial online] 2011 [cited 2019 Jun 24];54:654-5. Available from: http://www.ijpmonline.org/text.asp?2011/54/3/654/85139


Sir,

Since its development in 1998 by Kononen et al., [1] tissue microarray (TMA) is becoming a useful tool for research and quality control in immunohistochemistry (IHC) and in situ hybridization methods. [2] Array construction is done with the help of automated tissue arrayers, which are expensive and thus not suitable for laboratories in the developing world. Recently, different manual methods have been described for the development of inexpensive yet efficient microarrays. [3] Most of the techniques described, use a preconstructed paraffin donor block, into which holes are punched for inserting the tissue cores. [4] This is fraught with many difficulties, such as block breakage during punching, nonalignment of holes and mismatched length of the tissue cores and recipient holes. With the desire to construct a costeffective TMA while overcoming the above-mentioned difficulties, we tried an easy method for the same without using prefabricated recipient blocks. This method, which is a modification of the one initially described by Ni Chen et al. [5] requires no special instruments and has proved to be quite effective and reproducible. The tissue arrays thus constructed can be smoothly sectioned using a standard microtome and performed for a panel of immunohistochemical study with satisfactory results.

A disposable, skin punch biopsy needle of 2-mm diameter was used for punching cores from the donor blocks. A long steel wire with a blunt end was modified into a stylet. The skin biopsy needle needs no further modification, has a cylindrical core that ensures uniform tissue sampling and a sufficiently sharp cutting edge. Double-sided adhesive tape, a pair of stainless steel forceps, paper cutters, a 24 × 24 × 12 cmm embedding mould (Leica, Heidelberg, Germany) and plastic cassettes were the basic materials needed. A piece of double-sided adhesive tape was cut exactly according to the size of the embedding mold to provide a uniform cutting surface. The top surface was exposed to receive the tissue cores. The sites for attachment were marked with the help of a ruler and a felt pen, for better core alignment. The area to be cored was marked on the H and E-stained sections and then on the block by super imposing the marked slide on it. The skin punch biopsy needle was drilled into the donor block at the selected site, taking care that its approach was perpendicular for optimum tissue sampling [Figure 1]a. After pulling out the needle, the tissue core was delivered with help of a stylet and transferred to the tape with the help of forceps (cutting surface facing downwards) [Figure 1]b. The tissue cores were arranged in the predecided format. An unrelated core from any normal tissue (e.g., appendix) was attached at the starting point for orientation. With this technique, 5 × 7 to 7 × 7 arrays could be designed, yielding a total of 35 and 49 cores, respectively, per slide. The array platform with the tissue cores in situ was then transferred to the stainless steel mould [Figure 1]c. Melted paraffin was gently poured in to the mould from the sides, taking care to avoid tipping of the cores. Also 2 mm diameter cores have a sufficiently broad base. To ensure gradual and uniform setting of the block, the mould was initially cooled at room temperature and then transferred to a refrigerator for 20 min. After the block was evenly set, the adhesive tape was peeled off to expose the cutting surface [Figure 1]d. From the blocks thus prepared, 3 mm sections were cut on a rotary microtome (LEICA) and stained by routine H and E stains [Figure 2]a. Sections were also taken on poly L lycine coated slides for performing immunohistochemistry. We have successfully performed validation studies of estrogen receptor (ER) and progesterone receptor (PR) markers by comparing the immunoreactivity of whole sections with that of the microarrays [Figure 2]b. This technique has also been used to produce control slides containing positive as well as negative controls. Sections have been obtained till the loss of one complete tissue core. We could thus prepare 15-20 sections from each core. This technique is readily reproducible and does not require specialized personnel training. One technical disadvantage was the less number of sections prepared from each block. This was due to using archival blocks, which had already been sectioned multiple times, leading to thinner tissue cores.
Figure 1: Procedure of constructing a microarray, (a) Taking out a tissue core from the donor block with the skin biopsy needle, (b) Attaching the core to the adhesive tape, (c) The cores platform in an embedding mold, (d) The final paraffin block (e) The fi nal H and E stained slide from the recipient paraffin block

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Figure 2: (a) Tissue array stained with routine H and E, with a representative section of carcinoma breast, (b) Immunostaining for estrogen receptor (ER; Dako, clone-SP1) of the array slide

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Our experience with this novel technique indicates that this method is easy, cost effective, and offers a huge potential in laboratories with minimal technical sophistication and financial limitations.

 
   References Top

1.Kononen J, Bubendorf L, Kallioniemi A, Barlund M, Schraml P, Leighton S, et al. Tissue micro-arrays for high-throughput molecular profiling of tumor specimens. Nat Med 1998;4:844-7.  Back to cited text no. 1
    
2.Packeisen J, Buerger H, Krech R, Boecker W. Tissue microarrays: A new approach for quality control in immunohistochemistry. J Clin Pathol Mol Pathol 2002;55:613-5.   Back to cited text no. 2
    
3.Pires AR, Andreiuolo Fda M, de Souza SR. TMA for all: A new method for the construction of tissue microarrays without recipient paraffin block using custom-built needles. Diagn Pathol 2006;1:14.  Back to cited text no. 3
[PUBMED]  [FULLTEXT]  
4.Singh DK, Sakhuja P, Gondal R. Making and using inexpensive manually constructed tissue micro-array: Experience of a tertiary care hospital in India. Indian J Pathol Microbiol 2009;52:304-9.   Back to cited text no. 4
[PUBMED]  Medknow Journal  
5.Chen N, Zhou Q. Constructing tissue microarrays without prefabricating recipient blocks: A novel approach. Am J Clin Pathol 2005;124:103-7.  Back to cited text no. 5
[PUBMED]  [FULLTEXT]  

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Correspondence Address:
Gayatri S Pathak
S. No. 49/1 and 53/2, Department of Pathology, Shrimati Kashibai Navale Medical College and General Hospital, Narhe, Off Pune-Mumbai Bypass, Pune
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0377-4929.85139

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