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A new integrated concept for the improved preparation of sections of fresh or frozen tissue for light microscope histochemistry

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Summary

In order to obtain thin sections of plant tissues which combined good morphological preservation and the preservation of the substances and enzyme activities in the tissues, a concept of section preparation by external stabilization was developed. The main components are as follows: (1) appropriate supporting medium; (2) surface coating before each sectioning process, the coating being either non-permanent, permanent, or semi-permanent; (3) suitable techniques for affixing the coated sections to the slides using either pressure-sensitive adhesive or solvent-based adhesive; and (4) mounting media with defined refractive indices (preferably UV-curable, water-soluble monomers). By this approach, sections exhibiting excellent morphological and physiological preservation were obtained using either a cryostat at −30°C or a rotary microtome at room temperature.

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References

  • Altman FP (1971) The use of a new grade of polyvinyl alcohol for stabilising tissue sections during histochemical incubations. Histochemie 28:236–242

    Google Scholar 

  • Ashford AE, Allaway WG, McCully ME (1972) Low temperature embedding in glycol methacrylate for enzyme histochemistry in plant and animal tissue. J Histochem Cytochem 20:986–990

    Google Scholar 

  • Bazer GT, Knight DS (1980) An inexpensive vibrating microtome for sectioning fixed tissue. Stain Technol 55:39–42

    Google Scholar 

  • Boll HU, Reb R, Taugner R (1974) Synthetic coating: An improvement in ultracryotomy. Experientia 30:1103–1104

    Google Scholar 

  • Bonga JM (1961) A method for sectioning plant material using cellulose tape. Can J Bot 39:729

    Google Scholar 

  • Brandrup J, Immergut EH (ed) (1975) Polymer handbook. 2nd edn. John Wiley & Sons, New York

    Google Scholar 

  • Branton D, Ruch F, Waldner H (1964) Sectioning fresh plant material with a microtome. Stain Technol 39:250–251

    Google Scholar 

  • Bush V, Hewitt RE (1952) Frozen sectioning. A new and rapid method. Am J Pathol 28:863–867

    Google Scholar 

  • Clouser TE (1977) Tissue bath improvements for the Oxford vibratome. Stain Technol 52:319–322

    Google Scholar 

  • Cole MB, Sykes SM (1974) Glycol methacrylate in light microscopy: A routine method for embedding and sectioning animal tissues. Stain Technol 49:387–393

    Google Scholar 

  • Elder HY, Gray CC, Jardine AG, Chapman JN, Biddlecombe WH (1981) Optimum conditions for cryoquenching of small tissue blocks in liquid coolants. J Microsc 126:45–61

    Google Scholar 

  • Fasano AV, Berendsen PB, Labriola A (1974) The use of BEEM capsules in the preparation of fresh frozen cryostat sections. Stain Technol 49:53–55

    Google Scholar 

  • Fink S (1983) Histologische und histochemische Untersuchungen an Nadeln erkrankter Tannen und Fichten im Südschwarzwald. Allg Forstzeitschr 38:660–663

    Google Scholar 

  • Fink S (1986a) Some new methods for affixing sections to glass slides. II. Organic solvent-based adhesives. (in preparation)

  • Fink S (1986b) Some new methods for affixing sections to glass slides. III. Pressure-sensitive adhesives. (in preparation)

  • Fink S (1986c) An extended approach to the use of water-soluble monomers for the preparation of tissues for light microscopy. (in preparation)

  • Fitz-William WG, Jones GS, Goldberg B (1960) Cryostat techniques: methods for improving conservation and sectioning of tissue. Stain Technol 35:195–204

    Google Scholar 

  • Franks F (1977) Biological freezing and cryofixation. J Microsc 111:3–16

    Google Scholar 

  • Franks F, Asquith MH, Hammond CC, Skaer HB, Echlin P (1977) Polymeric cryoprotectants in the preservation of biological ultrastructure. J Microsc 110:223–270

    Google Scholar 

  • Fujita M, Harada H (1980) A simple embedding and lining method for sectioning of wood and wood-based materials using an alpha-cyanoacrylate resin. Bull Kyoto Univ Forest 52:216–220

    Google Scholar 

  • Gahan PB, McLean J, Kalina M, Sharma W (1967) Freezing-sectioning of plant tissues: the technique and its use in plant histochemistry. J Exp Bot 18:151–159

    Google Scholar 

  • Gatenby JB, Cowdry EV (1928) Bolles Lee's microtomist's vademecum. 9th edn. J & A Churchill, London

    Google Scholar 

  • Gerrits PO, Zuideveld R (1983) The influence of dehydration media and catalyst systems upon the enzyme activity of tissues embedded in 2-hydroxyethyl methacrylate: an evaluation of three dehydration media and two catalyst systems. Mikroskopie 40:321–328

    Google Scholar 

  • Gray P (1954) The microtomist's formulary and guide. Constable & Co, London

    Google Scholar 

  • Henking H (1886) Technische Mitteilungen zur Entwicklungsge-schichte. Z Wiss Mikrosk 3:470–479

    Google Scholar 

  • Hibben CR (1968) A simplified carbowax embedding technique for sectioning fresh leaves of woody plants. For Sci 14:298–300

    Google Scholar 

  • Higuchi S, Suga M, Dannenberg AM Jr, Schofield BH (1979) Histochemical demonstration of enzyme activities in plastic and paraffin embedded tissue sections. Stain Technol 54:5–12

    Google Scholar 

  • Jensen WA (1962) Botanical histochemistry. WH Freeman, San Francisco

    Google Scholar 

  • Kauer G (1984) Herstellung sehr dünner Paraffinschnitte mit Hilfe einer Klebebandmethode. Mikrokosmos 73:17–23

    Google Scholar 

  • Knox RB (1970) Freeze-sectioning of plant tissues. Stain Technol 45:265–272

    Google Scholar 

  • Kushida H (1961) A new embedding method for ultrathin sectioning using a methacrylate resin with three dimensional polymer structure. J Electron Microsc 10:194–199

    Google Scholar 

  • Kushida H, Fujita K (1975) Improved methods for embedding with water-miscible methacrylates. J Electron Microsc 24:175–176

    Google Scholar 

  • Läuchli A (1966) Cryostat technique for fresh plant tissues and its application in enzyme histochemistry. Planta 70:13–25

    Google Scholar 

  • Mark EL (1885) Notes on section cutting. Am Natural 19:628–631

    Google Scholar 

  • Maxwell A, Ritchie JSD (1971) A comparison of adhesives useable in the cold for the preparation of cryostat sections. Stain Technol 46:167–169

    Google Scholar 

  • Mayahara H, Fujimoto K, Noda T, Tamura I, Ogawa K (1981) The “microslicer”, a new instrument for making non-frozen sections. Acta Histochem Cytochem 14:211–219

    Google Scholar 

  • McLane SR Jr (1951) Higher polyethylene glycols as a water solutble matrix for sectioning fresh or fixed plant tissues. Stain Technol 26:63–64

    Google Scholar 

  • Namba M, Dannenberg AM Jr, Tanaka F (1983) Improvement in the histochemical demonstration of acid phosphatase, β-galactosidase and nonspecific esterase in glycol methacrylate tissue sections by cold temperature embedding. Stain Technol 58:207–213

    Google Scholar 

  • Parameswaran N, Fink S, Liese W (1985) Feinstrukturelle Untersuchungen an Nadeln geschädigter Tannen und Fichten aus Waldschadensgebieten im Schwarzwald. Eur J For Pathol 15:168–182

    Google Scholar 

  • Persidsky MD (1953) A vibratory microtome for sectioning living tissue. J Lab Clin Med 42:468–471

    Google Scholar 

  • Ruddell CL (1967a) Hydroxyethyl methacrylate combined with polyethylene glycol 400 and water; an embedding medium for routine 1–2 micron sectioning. Stain Technol 42:119–123

    Google Scholar 

  • Ruddell CL (1967b) Embedding media for 1–2 micron sectioning. 2. Hydroxyethyl methacrylate combined with 2-butoxyethanol. Stain Technol 42:253–255

    Google Scholar 

  • Rydberg H (1955) A method for microtomy of difficult meterials. Z Wiss Mikrosk 62:341–342

    Google Scholar 

  • Sims B (1974) A simple method of preparing 1–2 μm sections of large tissue blocks using glycol methacrylate. J Microsc 101:223–227

    Google Scholar 

  • Smith RE (1970) Comparative evaluation of two instruments and procedures to cut nonfrozen sections. J Histochem Cytochem 18:590–591

    Google Scholar 

  • Snodgrass MJ, Peterson RG (1969) An inexpensive microtome attachment for cutting 50-micron nonfrozen sections for electron microscopic histochemistry. Stain Technol 44:151–154

    Google Scholar 

  • Stibane FA (1984) Eine verbesserte Tesafilmmethode für die Histologie. Mikrosmos 73:180–182

    Google Scholar 

  • Taugner R, Supply P, Braun A, Droh R, Mahn J (1963) Zur Gefrierschnitt-Autoradiographie am ganzen Tier. Nucl Med 3:397–405

    Google Scholar 

  • Turner JG, Novacky A (1973) Mounting and quenching thin leaves for cryostat sectioning. Stain Technol 48:263–266

    Google Scholar 

  • Vanden Born WH (1963) Histochemical studies of enzyme distribution in shoot tips of white spruce (Picea glauca (Moench) Voss). Can J Bot 41:1509–1527

    Google Scholar 

  • Weaver GM, Layne REC (1965) Cryostat sectioning of woody plant materials. Can J Bot 43:478–481

    Google Scholar 

  • Wedeen RP (1969) Autoradiography of freeze-dried sections: studies of concentrative transport in the kidney. In: Roth LJ, Stumpf WE (ed) Autoradiography of diffusible substances. Academic Press, New York, pp 147–160

    Google Scholar 

  • Wills GD, Briscoe GA (1970) A pectin — agar — sucrose tissue support for cryostat sectioning of fragile plant tissue. Stain Technol 45:92–93

    Google Scholar 

  • Wohlrad F (1971) Möglichkeiten der Herstellung von Gewebsschnitten ohne Frierprozeß und Einbettung des Gewebes. Mikroskopie 27:80–84

    Google Scholar 

  • Ziesmer C (1957) Das Schneiden großer Paraffinblöcke mit Hiffe von Tesafilm. Z Wiss Mikosk 63:236–237

    Google Scholar 

Download references

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  1. Institut für Forstbotanik und Holzbiologie, Albert-Ludwigs-Universität, Bertoldstrasse 17, D-7800, Freiburg im Breisgau, Germany

    S. Fink

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Fink, S. A new integrated concept for the improved preparation of sections of fresh or frozen tissue for light microscope histochemistry. Histochemistry 86, 43–52 (1986). https://doi.org/10.1007/BF00492344

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