Tissue printing

Cover photo Tissue print of okra fruit stained with acidic silver nitrate for ascorbic acid localization. Photo courtesy of Dr. Rosannah Taylor. 

Kaufmann A, Koenig R, Lesemann D-E. Tissue print-immunoblotting reveals an uneven distribution of beet necrotic yellow vein and beet soil-borne viruses in sugar beet. Arch Virol 1992 126 329-335. 

A breakthrough for tissue printing, at least for the botanical sciences, came when Cassab and Varner (3) combined the use of nitrocellulose and antibody technology. They placed sections of a freshly cut soybean seed on nitrocellulose membranes and probed the resulting imprints using specific antibodies. They were able to show that soluble extensin protein is primarily localized in the seed coat and vascular tissues. 

Actually doing tissue printing is the best way to understand its simplicity (Fig. 1). Wide variations in the technique are possible and can be developed easily and quickly according to need. The following protocol for using tissue prints to detect tissue distribution of IAA-protein con-juguates in okra fruit details the immunoblotting technique as follows. 

Summary of Some Recent Applications of Tissue Printing... 

Soft tissue Cryostat tissue printing Conley and Hanson (6)... 

Sporophytic maize tissues Biochemical and tissue print analyses of hydroxyproline-rich glycoproteins Hood et al. (11)... 

Tissue prints on nitrocellulose membranes (Schleicher Schuell) (0.45 pm pore size) were first stained for protein using Ponceau S (Fig. 2A,B) and photographed immediately with T-max 100 black-and-white film. The membranes were then washed in PBS to remove the stain, and incubated for 2 h with shaking in blocking solution. 

Fig. 2. Tissue-print immunoblots of cross sections from okrafruitpods. Ponceau S protein staining of blots before immunodetection reveals the anatomical detail of sections subsequently probed with control serum (A) or with Phaseolus vulgaris IAA-peptide antibody (B). Arrows indicate the position of seeds remaining within the cross-sectioned pods.

Fig. 2. (Continued). (C) Control protein tissue prints with antibody omitted. (D) Tissue-prints incubated with Phaseolus vulgaris IAA peptide antibody. Crossreaction visualized by chemiluminescence. Arrows indicate strong localization of the IAA-peptide in seeds.

The reader is referred to ref. (35) for an overview of tissue printing and ref. (36) for a tissue-printing protocol for celery. 

Cassab GI, Varner JE. Immunocytolocalization of extensin in developing soybean seed coats by immunogold-silver staining and by tissue printing on nitrocellulose paper. J Cell Biol 1987 105 2581-2588. 

Antikainen M, Griffith M, Zhang J, Hon WC, Yang DSC, Pihakashi-Maunsbach K. Immunolocalization of antifreeze proteins in winter rye leaves, crowns, and roots by tissue printing. Plant Physiol 1996 110 845-857. 

Conley CA, Hanson MR. Cryostat tissue printing An improved method for histochemical and immunocytochemical localization in soft tissues. Biotechniques 1997 22 490-496. 

HoodKR, BaasiriRA, Fritz SE, Hood EE. Biochemical and tissue print analyses of hydroxyprohne-rich glycoproteins in cell walls of sporophytic maize tissues. Plant Physiol 1991 96 1214-1219. 

Roth R, Boudet AM, Pont-Lezica R. Lignification and cinnamyl alcohol dehydrogenase activity in developing stems of tomato and popular a spatial and kinetic study through tissue printing. J Exp Bot 1997 48 247-254. 

Taylor R, Varner JE. The role of calcium in maintaining cell-wall firmness studied by physical printing on nitrocellulose membranes, in Tissue Printing Tools for the Study of Anatomy, Histochemistry, and Gene Expression (Reid P, Pont-Lezica R, del Campillo E, Taylor R, eds.), Academic Press, New York, 1992, pp. 15-17. 

Taylor R, Inamine G, Anderson JD. Tissue printing as a tool for observing immunological and protein profiles in young and mature celery petioles. Plant Physiol 1993 102 1027-1031. 

B.N., Genega, E.M., Rubin, M.A., and Lenkinski, R.E. (2005) Tissue-print and print-phoresis as platform technologies for the molecular analysis of human surgical specimens Mapping tumor invasion of the prostate capsule. Nat. Med. 11 95-101. 

Latham, G., Magotra, A., and Gaston, S.M. (2006) Cancer biomarker quantification using RNA extracted from tumor tissue print micropeels. Ambion TechNotes 13(2) 25-26 www.ambion.com/techlib. 

Anionic direct dyes are the most widely used colourant within the tissue, printing and writing paper sectors. Having a typical molecular weight of 550-2,000, they produce a wide range of shades, although certain bright shades cannot be achieved... 

Tissue printing (pressure) This procedme allows the transfer of proteins to nitrocellulose membranes when a fresh cut organ or the cross sections of seeds, stems, tubers, leaves, and fruits are pressed against the membrane surface. This procedure was further modified to detect plant virus coat proteins on whole leaf blots. In this procedure, plant leaves to be blotted were placed on dry nitrocellulose membrane. The leaf and the nitrocellulose membrane were sandwiched on either side with blotting paper or one 0.75 inch thick block of plywood. The sandwich was positioned into a Carver laboratory press, and 10 000 psi of pressure was applied for 2 min. After disassembly, the membrane was air-dried for 20 min prior to standard immunodetection procedures. This procedure was shown to transfer protein uniformly. Some amount of chlorophyll was found to be transferred along with the protein, but this was not found to interfere significantly with the subsequent visualization of results. 

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