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

Abstract

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: appropriate supporting medium; surface coating before each sectioning process, the coating being either non-permanent, permanent, or semi-permanent; suitable techniques for affixing the coated sections to the slides using either pressure-sensitive adhesive or solvent-based adhesive; and 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 degrees C or a rotary microtome at room temperature.

Cytochemical Measurement of Lysosomal Responses in the Digestive Cells of Wild Pacific Oyster, Crassostrea gigas on the South Coast of Korea

“…그러나 조직관찰을 위한 파라 핀 블록 제작은 그 준비 기간이 오래 걸리며, 고정과 포매 과정에서 지질성분 혹은 효소의 활성을 잃어버리는 단점 이 있다 (Fink 1986 (Fink 1986;Lowe 1988;Moore 1988;Krishnakumar et al 1994;Dimitriadis et al 2004). 이와 같은 이유로, 대부분의 해산 이매패류 소화맹 낭 리소솜 활성의 연구는 동결절편 된 조직에서 세포화학 적인 방법을 이용하여 이루어지고 있다 (Lowe 1988;Moore 1988;Krishnakumar et al 1994;Petrovi et al 2001;Kagley et al 2003;Dimitriadis et al 2004).…”

Some New Methods for Affixing Sections to Glass Slides. Ii. Organic-Solvent Based Adhesives

“…These tests included prolonged boiling in water, exposure to acids and alkalis, exposure to various solvents at room or elevated temperatures. Other slides with sections were routinely stained with various stains, passed through alcohol and xylene, and then mounted with the usual resinous media or with polymerizable monomers (Fink 1986).…”

“…This was where Fitz-William et al (1960) and Yos (196 1) already had their problems, using rubber solutions for affixing the sections. This can be overcome either by using an adhesive insoluble in xylene (cellulose nitrate, cellulose acetate phthalate, cyanoacrylate adhesive) or avoiding the step with xylene and using aqueous mounting media, alcoholmiscible media (e.g., "Euparal") or preferably polymerizable monomeric media (Fink 1986). …”

Some New Methods for Affixing Sections to Glass Slides. III. Pressure-Sensitive Adhesives

“…Most other sections were first either stained according to different formulas or incubated for 12 h r at 37 C to demonstrate acid phosphatase (Jensen 1962). T h e slides were then either mounted directly with glycerine jelly, or first dehydrated with alcohol, passed through xylene, and mounted with Entellan (Merck) or Euparal, or they were dehydrated with GMA or PEGDMA and mounted using the same monomers as polymerizable mounting media (Fink 1986 and manuscript in preparation). Refractive indices were measured with an Abbe refractometer (Zeiss) and compared with the values from Brandrup and Immergut (1 975).…”