Venetian Turpentine as an Aid in Squashing and Concomitant Production of Durable Chromosome Mounts

Root tips are hydrolyzed in 1 N HCl at 60 C for 10-12 min, Feulgen stained, and macerated in a minimal amount of propiono-carmine. A drop of Venetian turpentine mounting medium (Harleco brand was used) is mixed with the carmine stain, the cover slip applied, the tissue pressed gently while observed under a dissecting microscope to spread the chromosomes, and finally firmly squashed. High quality slides of over 1 yr durability are obtained which are well suited to morphological studies, photographing under oil, or routine screening in polyploidy studies. The carmine stain in conjunction with the Feulgen aids to give contrast to chromosomes which are difficult to stain (e.g. hops) but it may be omitted for other species.     

Xylene-Cellulose Caprate as a Histologic Mounting Medium of Low Refractive Index

A 50% solution of cellulose caprate in xylene forms a clear pale yellowish syrupy solution of proper consistency for mounting stained and unstained sections. Cover slips become immovable in 30 minutes and slides do not stick together after one-day drying. The refractive index of the xylene solution is 1.4860, of the dry resin 1.4734, and unstained sections remain easily visible. Most stains are well preserved. Prussian blue is well preserved, cobalt sulfide decolorizes in a month.     

Advantages of Epoxy Resin as a Mounting Medium for Light Microscopy

The need for very durable mounting is especially felt in the teaching of parasitology and mycology; otherwise, the availability of microscope slides may depend on the use of fresh specimens. Resinous mounts and those in aqueous media sealed with fingernail lacquer, paraffin or asphalt do not preserve specimens satisfactorily. Polyvinylpyrrolidone (pvp), a water-soluble mounting medium described by Burstone (1962), cannot be applied directly for mounting of insects and certain other parasites which have water-repelling integuments; moreover, pvp bleaches eosin. Grimley et al. (1965) prepared large epoxy sections of tissues from which areas for electron microscopy could be selected. This procedure however is designed for electron microscopic techniques whereas the present paper describes a direct epoxy mounting method to produce permanent mounts for light microscopy.     

Euparal as A Mounting Medium for Preserving Fluorescence of Aniline Blue in Plant Material

Keeping quality of aniline blue-stained plant material can be greatly enhanced by substituting Euparal for aqueous aniline blue as the mounting medium. Pollen tubes which have been stained for other purposes (e.g. nuclei and chromosomes) and mounted in resinous medium can be restained with aniline blue and fluorescence of pollen tubes can still be observed.     

Permanent Mounting of Unstained and Aceto-Orcein Stained Cells in the Water-Soluble Medium,Abopon

For cellular morphology, mammalian cells were grown on cover slips in Leigh ton tubes, fixed in 1% osmic acid vapor for 2 min, decolorized with 30% H₂O₂ in 5% ammonium oxalate solution (1:7) for 2 min, then washed thoroughly, and finally mounted in a water-soluble medium consisting of a saturated solution of Abopon in 0.2 M phosphate buffer, pH 7.0. For chromosomal analysis of similarly cultured cells, aceto-orcein preparations were made by conventional methods, with the following minor modifications: following pretreatment with colchicine and hypotonic expansion, the cells on the cover slips were fixed in acetic-alcohol (1:3), air dried, incubated at 37° C for 15 min in 2% orcein in 45% acetic acid, rinsed in 45% acetic acid, washed several times in distilled water, and finally mounted in Abopon mounting medium. Both kinds of preparations were allowed to harden for 24 hr before being handled. Such slides will keep for years at room temperature. Studies requiring frequent comparisons of cellular and chromosomal morphology of cultured cells can thus be extended over long periods of time.     

The Piccolyte Resins as Microscopic Mounting Media

A new series of synthetic resins, the “Piccolytes” (β-pinene polymers), is recommended for permanent mounting media in histological work. These resins, which are available with various melting points, are of correct refractive indices, very low acid numbers, are pale, non-yellowing, have good adhesion to glass, and are freely soluble in xylene. Of a considerable variety of newer synthetic resins which were tried (stimulated by the recent scarcity of Canada balsam), only these terpene resins were found entirely satisfactory. Being of controlled manufacture, they are uniform in characteristics as well as readily available and very cheap.

A partial review of the literature of synthetic resin mountants is included.     

Staining and Mounting Helminths

The following procedure is recommended: Fix ces-todes and trematodes (while held flat between glass slides) 0.5-2.0 hr. in the following mixture: formalin, 15; acetic acid (gl.), 5; glycerol, 10; 95% ethyl alcohol, 24; distilled H₂O, 46; all proportions by volume. After freeing them from the slides, wash thoroughly in running water and stain immediately thereafter. Stock staining solution: ferric ammonium alum (violet cryst.), 2 g.; distilled H₂O (cold) 100 ml.; after solution, add 2 ml. concentrated H₂SO₄, bring to a boil; add 1 g. coelestin blue B (Nat. Aniline), boil 3-5 min.; cool and add 10 ml. absolute methyl alcohol and 10 ml. glycerol. Dilute 1 vol. with 3 vol. distilled H20 for use. Stain 5-30 min., depending on size of specimens. Wash with 2 changes 0.5 hr. each of distilled H₂O, then 50% isopropyl alcohol 12-16 hr., 50% isopropyl alcohol 2 hr., followed by graded isopropyl alcohol for dehydration. Ether: ethyl alcohol (equal parts), 1 hr., is followed by embedding in celloidin in a sheet just thick enough to cover the specimens. Trim embedded specimens and dehydrate with isopropyl alcohol, 80%, 90% and absolute. Clear in beechwood creosote. Mount in balsam with cover glasses that overlap the edges of the celloidin 1-2 mm. While drying at 37°C, refill edges of mount with fresh balsam as needed. When dry, remove excess balsam and ring the edges with ordinary gloss enamel paint.     

Staining and Mounting Helminths

The following procedure is recommended: Fix ces-todes and trematodes (while held flat between glass slides) 0.5–2.0 hr. in the following mixture: formalin, 15; acetic acid (gl.), 5; glycerol, 10; 95% ethyl alcohol, 24; distilled H₂O, 46; all proportions by volume. After freeing them from the slides, wash thoroughly in running water and stain immediately thereafter. Stock staining solution: ferric ammonium alum (violet cryst.), 2 g.; distilled H₂O (cold) 100 ml.; after solution, add 2 ml. concentrated H₂SO₄, bring to a boil; add 1 g. coelestin blue B (Nat. Aniline), boil 3–5 min.; cool and add 10 ml. absolute methyl alcohol and 10 ml. glycerol. Dilute 1 vol. with 3 vol. distilled H20 for use. Stain 5–30 min., depending on size of specimens. Wash with 2 changes 0.5 hr. each of distilled H₂O, then 50% isopropyl alcohol 12–16 hr., 50% isopropyl alcohol 2 hr., followed by graded isopropyl alcohol for dehydration. Ether: ethyl alcohol (equal parts), 1 hr., is followed by embedding in celloidin in a sheet just thick enough to cover the specimens. Trim embedded specimens and dehydrate with isopropyl alcohol, 80%, 90% and absolute. Clear in beechwood creosote. Mount in balsam with cover glasses that overlap the edges of the celloidin 1–2 mm. While drying at 37°C, refill edges of mount with fresh balsam as needed. When dry, remove excess balsam and ring the edges with ordinary gloss enamel paint.     

梁河脂松节油、脂松香的物理和化学特征

研究梁河脂松节油的相对密度、折光率、初馏点、馏程体积、旋光度和化学组成以及脂松香的软化点、酸值、不皂化物含量、乙醇不溶物、灰分、旋光度和化学组成等物理和化学特征。结果表明:梁河的脂松节油在化学组成上因含有大量3-蒈烯导致其蒎烯含量低于GB/T 12901-2006,其物理指标中的折光率偏高。梁河的脂松香因化学组成中含有大量中性物,致其物理指标中酸值偏低,不皂化物含量偏高,超出GB/T 8145-2003。鉴于3-蒈烯这一特殊资源,有必要制定高3-蒈烯脂松节油标准。     

An Improved Squash Technique for Human Male Meiotic Chromosomes: Softening and Concentration of Cells; Mounting in Hoyer's Medium

Human testicular material obtained from autopsies was processed as follows: (1) swelled with 0.3% sodium citrate 1-6 hr; (2) softened with 3 M glucono-delta-lactone 2 hr; (3) stained with aceto- or propiono-carmine overnight; (4) washed in 70% alcohol; (5) macerated in 1:1 alcohol-acetic acid; (6) filtered through gauze to remove tubules and connective tissue; (7) filtrate centrifuged to separate spermatocytes from debris; (8) a drop of supernate from just above the precipitate, containing stained cells, mixed with Hoyer's medium; (9) cover slip applied, preparation warmed, and (10) squashed. Chromosomes remained in good condition for 8 mo.     

The Function of Mounting Behaviour in Farmed Red Deer Calves

The function of mounting behaviour was studied in farmed red deer (Cervus elaphus) calves. On the basis of previous work, we tested two alternative hypotheses about the function of this behaviour. The first hypothesis deals with the proximate function of the behaviour. Three predictions were tested: (1a) mounting behaviour attracts the attention of the mother and/or consolidates the mother–calf bond; (1b) mounting is intended to obtain more milk; (1c) mounting is intended to prevent other calves from sucking from the mother. The second hypothesis deals with an ultimate function in practising for future life with two predictions tested: (2a) mounting behaviour is part of calves' play behaviour promoting development of the calf's locomotor and social skills; (2b) calves gain sexual experience through mounting behaviour. For the study, 50 hind‐calf pairs were observed. The maternity of individual hinds was confirmed by a genetical analysis. The hinds were classified as ‘maternal’ and ‘non‐maternal’ and the calves as ‘filial’ and ‘non‐filial’. We recorded 40 cases of mounting behaviour involving 25 hinds and 21 calves. Our results suggest that the mounting behaviour of red deer calves serves several different functions depending on the circumstances. In association with suckling, calves of both sexes mounted maternal hinds mainly to attract their attention and to achieve another suckling. In situations not associated with suckling, mounting by male calves might be considered part of their sexual training. On the other hand, mounting by female calves probably reflects the attempt to maintain contact with their mother in tense situations.