Card Mounts for Handling Root Tips in the Paraffin Method

A method is described for utilizing card mounts to facilitate the handling of root tip samples and similar material in the paraffin method. The freshly collected roots are attached to temporary card mounts having dimensions of approximately 2 cm. × 2.5 cm. with DuPont Household Cement or a similar adhesive that hardens rapidly in the ordinary aqueous fixing fluids and is insoluble in the lower grades of alcohol. After fixation and dehydration to 75% alcohol, the roots are transferred to permanent card mounts on which they are carefully oriented for sectioning. Mucilage or glue which hardens rapidly in 85% alcohol and is insoluble in the ordinary dehydrating and infiltrating media is used in making the permanent card mounts. Detailed instructions are given for preparing and handling the card mounts, and a system of labeling the mounts is also suggested.     

A Method of Permanently Mounting Biological Tissue Cleared in Herr's Four-And-A-Half Clearing Fluid

A technique which should be generally applicable for preparing permanent mounts of tissue cleared in Herr's four-and-a-half clearing fluid is described. This technique involves transferring plant or animal tissues through a series of solutions consisting of Pienarr's fixative, Herr's clearing fluid, chloral hydrate, acetone and finally polyester resin for mounting. Material prepared using this method is exceptionally transparent and well preserved, and is suitable for either phase contrast or Nomarski interference microscopy.     

Laboratory Hints from the Literature: A Department Devoted to Abstracts of Books and Papers from Other Journals Dealing with Stains and Microscopic Technic in General

A technique which should be generally applicable for preparing permanent mounts of tissue cleared in Herr's four-and-a-half clearing fluid is described. This technique involves transferring plant or animal tissues through a series of solutions consisting of Pienarr's fixative, Herr's clearing fluid, chloral hydrate, acetone and finally polyester resin for mounting. Material prepared using this method is exceptionally transparent and well preserved, and is suitable for either phase contrast or Nomarski interference microscopy.     

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.     

Mounting Media for phase Microscope Specimens

-Results with phase microscopy can often be improved by the proper selection of mounting media. Temporary mounts can first be made in liquid mixtures of water and glycerol, butyl carbitol and alpha-chloronaphthalene, alpha-chloronaphthalene and alpha-bromonaphthalene or alpha-bromonaphthalene with methylene iodide to determine what index liquid serves best to emphasize structures of primary interest. Once this has been determined, permanent mounts can be prepared by the substitution of a solid mounting medium that most closely approaches the refractive index of the liquid.     

A coverslip culture technique for preparing permanent fungus mounts

A coverslip culture technique for the growth of fungi, with subsequent preparation of permanent mounts for microscopic examination, is described. This simplified technique allows examination at different stages of development of the fungus, or can be used in the preparation of large numbers of similar mounts for teaching purposes.     

A coverslip culture technique for preparing permanent fungus mounts

A coverslip culture technique for the growth of fungi, with subsequent preparation of permanent mounts for microscopic examination, is described. This simplified technique allows examination at different stages of development of the fungus, or can be used in the preparation of large numbers of similar mounts for teaching purposes.     

A review on the biosynthesis of metallic nanoparticles (gold and silver) using bio-components of microalgae: Formation mechanism and applications

The synthesis of nanoparticles (NP) using algae has been underexploited and even unexplored. In recent times, there are few reports on the synthesis of NP using algae, which are being used as a bio-factory for the synthesis. Moreover, the algae are a renewable source, so that it could be effectively explored in the green synthesis of NP. Hence, this review reports on the biosynthesis of NP especially gold and silver NP using algae. The most widely reported NP from algae are silver and gold than any other metallic NP, which might be due to their enormous biomedical field applications. The NP synthesized by this method is mainly in spherical shape; the reports are revealing the fact that the cell free extracts are highly exploited for the synthesis than the biomass, which is associated with the problem of recovering the particles. Besides, mechanism involving in the reduction and stabilization is well demonstrated to deepen the knowledge towards enhancement possibilities for the synthesis and applications.     

Plant-Virus Differentiation by Trypan-Blue Reactions within Infected Tissue

Trypan blue has proved effective for demonstrating the presence of certain plant viruses within infected tissues. The amorphous and crystalline inclusions which constitute cytological evidence of viruses stain proportionately. The effects produced by different viruses react differently to the stain and those inclusions which do not absorb trypan blue tend to stain with phloxine. This selective staining is the basis for using trypan blue singly and in combination with phloxine as standardized procedures for demonstrating and differentiating cytological evidence of plant viruses. These tests are very rapid and are especially applicable to temporary mounts of living tissue but permanent mounts can be made from material fixed in formalin.     

From extracellular to intracellular: the establishment of mitochondria and chloroplasts.

Paracoccus and Rhodopseudomonas are unusual among bacteria in having a majority of the biochemical features of mitochondria; blue-green algae have many of the features of chloroplasts. The theory of serial endosymbiosis proposes that a primitive eukaryote successively took up bacteria and blue-green algae to yield mitochondria and chloroplasts respectively. Possible characteristics of transitional forms are indicated both by the primitive amoeba, Pelomyxa, which lacks mitochondria but contains a permanent population of endosymbiotic bacteria, and by several anomalous eukaryotic algae, e.g. Cyanophora, which contain cyanelles instead of chloroplasts. Blue-green algae appear to be obvious precursors of red algal chloroplasts but the ancestry of other chloroplasts is less certain, though the epizoic symbiont, Prochloron, may resemble the ancestral green algal chloroplast. We speculate that the chloroplasts of the remaining algae may have been a eukaryotic origin. The evolution or organelles from endosymbiotic precursors would involve their integration with the host cell biochemically, structurally and numerically.     

Permanent mounts of fungus cultures grown on media optimal for production of characteristic structures

Summary 1. Dermatophyte cultures in plastic flasks were compared on four different media for total growth, sporulation, and pigment production. Czapek's agar favored more rapid and more abundant sporulation in most cultures than Sabouraud's dextrose agar, potato dextrose agar, or wort agar.2. A simple technique for embedding agar island cultures of fungi is described. These plastic-embedded cultures form permanent mounts useful for study of microscopic morphology.3. A relatively durable slant culture in a similar plastic flask is suggested as a companion for study of gross morphologic features of the cultures.     

Durable Whole Mounts of Sebaceous Glands Colored with Oil Blue N

Studies of the distribution, abundance, size and shape of sebaeous glands are most readily made with relatively permanent whole mounts of skin samples which are: fixed in 10% formalin, washed, stained 8-16 hours with oil blue N (2 parts sat. oil blue N in 60% isopropanol: 1 part water), washed, dissected free of obscuring connective tissue, washed, and mounted on slides in glycerol gelatin. After 2 or 3 days, the edges of the cover glass are sealed with clear lacquer.     

Whole Mounts of Flat and Round worms for Morphological Studies

A method for making permanent whole mounts of flat and round worms is described. The specimens are mounted in a drop of acid fuchsin lacto-phenol on a slide and warmed for 6 hours at 60°C. The acid fuchsin is replaced by light cotton-blue (anilin blue, W. S.) in lacto-phenol, till the desired contrast is obtained. After this, the forms are mounted in pure lacto-phenol, using the coverglass. The margin of the coverglass is sealed with the sealing media devised by Dade and Waller (equal parts of damar balsam and beeswax)     

Effects of Fixation and Dehydration Procedures on Marine Nematodes

Different combinations of fixation and dehydration procedures for the preparation of permanent mounts of marine nematodes of the subfamily Oncholaiminae were tested and compared. Qualitatively, the best specimens resulted from Seinhorst''s killing method and fixation in FAA; the dehydration procedure was of less significance. Quantitatively, no significant modification of measurements resulted from any of the methods used. Sources of error in measurements are discussed.     

Lacto-Phenol Preparations

More or less permanent mounts of fungi, algae, root tips, epidermis, germinating spores, and other small objects may be made readily by transferring the material to Amann's lacto-phenol containing anilin blue, W. S. or acid fuchsin, used singly or mixed. The addition of 20 to 25% of glacial acetic acid to these mixtures is frequently advantageous; or material may be stained with various dyes—acid fuchsin, anilin blue, W. S. (cotton blue), rose bengal, phloxine, hematoxylin—in aqueous solutions containing 5% of phenol, and then mounted in lacto-phenol, 50% glycerin or phenolglycerin, depending on the dye used. The phenol solutions of acid fuchsin and anilin blue are acidified with acetic acid and those of rose bengal and phloxine are made slightly alkaline with ammonium hydroxide. The addition of ferric chloride to acid fuchsin or acidified hematoxylin may improve staining. Fixation may be preferable but may be omitted, especially with fungi. Formulae for the mounting media and ten staining mixtures are given.     

A Technique for C-banding Chromosomes with Pinacyanol Chloride

A technique is presented for rapid C-banding of eukaryotic chromosomes with pinacyanol chloride. This technique has given excellent definition and clarity to chromosome bands in plant and animal chromosomes. In permanent mounts the chromosomes did not fade after storage for up to two years.     

123 Permanent Fluorescent Nuclear Staining of Binucleate Dinoflagellates

Typically, fluorescent microscopy of dinoflagellate nuclei is of poor resolution, due mainly to visual obstruction of the nuclei by plastids, pigment granules, and thecal plates. Moreover, the usual slide mounts using buffered glycerol are temporary, and fade after a week or so. We have developed a procedure to clear pigments from dinoflagellates, followed by fluorescent staining of the nuclei. The cells are then prepared as permanent mounts using an ultraviolet light-catalyzed resin to produce stained samples which may be kept for at least three years with little loss of fluorescence. This procedure can also be used to prepare plastic embedded dinoflagellate cells which can then be sectioned at 1–2 nm, fluorescent stained, and permanently mounted. Suitable nuclear stains are DAPI, Hoechst 33258, ethidium bromide and acridine orange. The dinoflagellate (dinokaryotic), and endosymbiont (eukaryotic) nuclei are clearly visualized, revealing individual chromosomes in the dinoflagellate nucleus, and a highly lobed morphology of the endosymbiont nucleus.     

Notes and queries

A method for making permanent whole mounts of flat and round worms is described. The specimens are mounted in a drop of acid fuchsin lacto-phenol on a slide and warmed for 6 hours at 60°C. The acid fuchsin is replaced by light cotton-blue (anilin blue, W. S.) in lacto-phenol, till the desired contrast is obtained. After this, the forms are mounted in pure lacto-phenol, using the coverglass. The margin of the coverglass is sealed with the sealing media devised by Dade and Waller (equal parts of damar balsam and beeswax)     

Laboratory sources of error for algal community attributes during sample preparation and counting

Applied algal studies typically require enumeration of preserved cells. As applications of algal assessments proliferate, understanding sources of variability inherent in the methods by which abundance and species composition data are obtained becomes even more important for precision of measurements. We performed replicate counts of diatoms on permanently fixed coverglasses and all algae in Palmer–Maloney chambers to assess precision and accuracy of measurements derived from common counting methods. We counted diatoms and all algae with transects and random fields. Variability estimates (precision) of diatom density, species diversity, and species composition on permanent coverglasses were low between replicate subsamples and between replicate transects. However, average density estimates of diatoms settled on coverglasses determined with transect methods were 42–52% greater than density estimates made with random fields. This bias was due to a predictable, nonrandom distribution of diatoms on the coverglass with few diatoms near edges. Despite bias in density when counting diatoms along coverglass transects, no bias was observed in estimates of species composition. Estimates of density and taxa richness of all-algae in Palmer–Maloney chambers also had low variability among multiple transects and high similarity in species composition between transects. In addition, counting method in Palmer–Maloney chambers did not affect estimates of algal cell density, taxa richness, and species composition, which suggested that counting units were distributed randomly in the chambers. Thus, most sources of variability in sample preparation and analysis are small; however, transect counts should not be used to estimate cell density, and sufficient numbers of random fields must be counted to account for edge effects on cell distribution with material settled on permanently fixed coverglasses.