A New Hematological Stain: I. Constituents and Methods of Use

The stain proposed by the author is quickly and simply prepared by mixing equal parts of the following permanent stock solutions:

The mixed stain is usable, for at least eight months, and is applicable to practically all hematological purposes: blood smears, fixed sections, frozen sections, and touch preparations. In the technics utilized to produce its action on preparations treated in different ways, the only variants are the methods for treating the cells, while the stain itself remains totally unchanged for all purposes. For blood smears fixed with methyl alcohol, the technic consists merely of pouring the stain on the slide, leaving it for 5 to 7 minutes, and then washing it off. On sections, a further process of differentiation with acid acetone is rapidly carried out. The various types of granules, including megakaryocyte and platelet granules, are clearly demonstrated. For frozen sections, the technic is extremely rapid, yet yields excellent differentiation.     

The Use of the Feulgen Technic with Certain Plant Materials

The Feulgen technic as modified by Heitz promises to become an extremely useful tool in the solution of certain cytological problems. A procedure is outlined for using this technic with root tip smears, and smears of plant microspores. The chief improvement suggested over previous methods is that the material be mounted in euparal, after immersion in 95% alcohol. The technic is of value in the study of chromosome fragmentation, chromatid coiling, centromeres, etc., in both somatic tissue and in microspores.     

Improvements of Hagmann's Method for Injecting Insect Tracheae

A simple technic for holding and releasing the specimen cage under vacuum by means of paraffin and heat is described. The technic is very easy to follow and greatly facilitates the handling of large numbers of specimens. The specimen cage used is illustrated and the use of a household detergent, Tide, in substitution of Santomerse for preparing Hagmann's solution A is mentioned.     

A simple R banding technic.

A simplified R banding technic is described which provides excellent delineation of major regions, easy identification of all chromosomes, and an accurate comparison of homologue lengths. The technic is simple, requiring only an initial incubation in buffer at 85 degrees C followed by acridine orange staining. The best presentation of the R banded chromosomes was obtained by printing in black and white from color transparency film. Variations in the length of the short arms of the acrocentric chromosomes are clearly and consistently defined. Satellites are not demarcated and appear as part of the short arm. Consistent banding was obtained, and the technic is suitable for use in routine clinical cytogenetic work.     

A Method for Analysis of Protein in Individual Amoebae

SYNOPSIS The microelectrophoresis technic of Hydén and Lange has been adapted for analysis of the proteins of individual Amoeba proteus, A. dubia , and Pelomyxa carolinensis. The method has the sensitivity required for analyzing electrophoretic mobilities of the proteins of these 3 species and is reproducible. The possibility of using this technic to differentiate amoeba species according to their protein composition is discussed.     

Observations on the Technic for Differential Staining of the Cell Cycle Using Safranin and Indigo-Picrocarmine

By means of auto-radiography and Feulgen-microspectrophotometry, a published technic using safranin and inago-picrocarmine was investigated for reliability in staining cells differentially in different stages of the cell cycle. The technic was found to be unreliable.     

A Flagella Staining Technic for Soil Bacteria

In an attempt to stain the flagella of soil bacteria, many of which have flagella so fine that they are hard to stain by most methods, a technic was developed which combines the best points of Hofer and Wilson's method with that of Bailey as developed by O'Toole. Satisfactory preparations have been obtained for organisms of the genera Pseudomonas, Phytomonas, Alcaligenes, Escherichia, Azotobacter, and Bacillus. This technic, therefore, is recommended as a rapid and constant method for routine flagella staining of all motile aerobic organisms; it combines Hofer and Wilson's method of cleaning the slides with O'Toole's technic of spreading the smears and with a modification of Bailey's mordant.     

A Carmine-Giemsa Staining Technic For Meiotic Prophase Chromosomes Of The Genus Beta L

For a detailed study of chromosome morphology in meiotic prophase stages of Beta species, a special double staining technic has been developed. It consists of combined maceration-staining in an ethanol-hydrochloric acid-carmine mixture followed by poststaining of the squashed material in a diluted Giemsa solution. The technic yields well-spread prophase meiotic nuclei showing detailed structures both in weaker stained chromosome segments and in threadlike chromatin structures. This technic proved to be especially favorable for stages which are difficult to interpret, such as pachytene, schizotene and diffuse stages.     

Technical note on the demonstration of rheumatoid factor by immunofluorescence

The authors present a new technic to point out the rheumatoid factor by immunofluorescence and compare this technic and the others to classical technics of agglutination. This technic allows to avoid the absorption of rheumatoid factor in the serological reactions using the immunofluorescence.     

Patterns of Cortical Stability in Tetrahymena*

SYNOPSIS. A simplified technic is presented for determining clonal stability patterns for numbers of ciliary rows in Tetrahymena. The technic involves plotting the variance of meridian number against the mean number of meridians. By this procedure strains of different syngens, difficult to discriminate on other grounds, may sometimes be shown to be distinctive.     

The Acetocarmine Smear Technic

The acetocannine smear technic is described primarily for beginners. An effort was made to bring together into one paper the work-a-day details to be followed in collecting specimens and making preparations of plant chromosomes at prophase, of condensed chromosomes at meiosis and mitosis (including mitosis in microspores), and of coiled chromonemata. A schedule for converting temporary slides into permanent mounts is given, and a technic for making smears of the chromosomes in the salivary glands of Drosophila is described.     

A carmine-Giemsa staining technic for meiotic prophase chromosomes of the genus Beta L.

For a detailed study of chromosome morphology in meiotic prophase stages of Beta species, a special double staining technic has been developed. It consists of combined maceration-staining in an ethanol-hydrochloric acid-carmine mixture followed by poststaining of the squashed material in a diluted Giemsa solution. The technic yields well-spread prophase meiotic nuclei showing detailed structures both in weaker stained chromosome segments and in threadlike chromatin structures. This technic proved to be especially favorable for stages which are difficult to interpret, such as pachytene, schizotene and diffuse stages.     

Polarized Light Technic A Comparison with the Marchi Method

The authors compare polarized light technic for the study of degeneration of myelinated nerves with the Marchi method. While polarized light shows much the same thing, it has the advantages of being much more rapid, more sensitive, and constant. It does not depend on fixation and staining but on the chemical structure of the myelin substance. These changes in structure begin before the third hour after cutting the nerve of a rat. No further technic is involved beside the making of frozen sections. Photographs are presented to illustrate the method.     

Manipulation of Tissues for Improved Aceto-Carmine Staining

The importance of the study of the structural-mechanic properties of any particular material in order to improve the technic for aceto-carmine smears and to obtain better preparations of that material has not been, perhaps, sufficiently emphasized in the large number of papers on such cytological technic. The usefulness of such a study will be shown here in two cases met by the writer.     

A Modification of Mayer's Mucihematein Technic

A relatively simple technic giving consistent results has been evolved from Mayer's mucihematein technic¹ by substituting hematoxylin for hematein and omitting the nitric acid. The hematoxylin is oxidized with sodium iodate (NaIO₃).

This modification is effective on the same types of mucin as Mayer's original mucihematein. With this modified technic, mucin stains a deep violet, cell nuclei pale gray blue, and connective tissue pale gray to colorless in tissues fixed in all the more common fixatives. The modified stain retains this selectivity for at least 200 days.     

The Weigert-Pal Technic for Staining Myelin

The writer gives a schedule for carrying out the Weigert-Pal technic by which three difficulties of the original technic are overcome: the tendency of the sections to become brittle; the difficulty of observing the extent of the reaction occurring in the permanganate solution; and the slowness with which the reaction takes place. By the method proposed as many as 300 sections may be stained and differentiated in the time it formerly took to handle 50.     

An Aceto-Carmine Squash Technic for Mature Embryo Sacs

A method is described by which, whole embryo sacs of Nicotiana, Petunia and no doubt of certain other genera can be obtained readily in aceto-carmine ovule squashes. Although application of the technic to megagametogenesis and fertilization stages is stressed in this paper, use of the method allows development to be traced from the archespore up to the second or third division of endosperm nuclei. The success of the technic depends on four phases:-1) fixation in a medium that causes cell and nuclear structures to become pliable, yet rigid enough that their spatial relationships are not greatly distorted in squashing; 2) heat, which apparently increases the cohesion of cytoplasmic and nuclear constituents; 3) maceration to separate the embryo sac from surrounding cells; and 4) the use of a stain that differentiates the various nuclear structures as well as those of the cytoplasm. Staining of the cytoplasm, essential in some embryological investigations, is one advantage of the aceto-carmine squash method over the Feulgen procedure. In contrast to the Feulgen ovule squash method the aceto-carmine technic will probably be most useful in genera having numerous small ovules. Advantages and defects of the aceto-carmine procedure as compared with the paraffin technic are discussed, likewise the possible usefulness of the former in studies of sterility and in certain other special connections.     

Phloxine As An Histologic Stain, Especially In Combination With Hematoxylin

A staining schedule employing phloxine as a counter-stain to Erlich's acid hematoxylin is presented. Fixation is best with Zenker's fluid, although formalin can be used. The technic is similar to the standard hematoxylin-eosin formulae but because of the staining advantages of phloxine over eosin, the technic is simpler, and quicker, resulting in clearly differentiated sections which do not fade as soon as do eosin-stained slides. A brief summary of the uses of phloxine as a biological stain is given and its advantages over eosin are discussed.     

Alcianblue/PAS or PAS/alcianblue? Remarks on a classical technic used in carbohydrate histochemistry

The combined alcian blue (AB)/PAS technic is widely used for the detection and characterization of mucosubstances in tissue sections. Mostly the sequence AB/PAS is used, occasionally also the reserved sequence PAS/AB. The present study shows clearly that the sequence of the combined technic, i.e. AB/PAS or PAS/AB is substantially influencing the results. So it could be demonstrated that by using the combination PAS/AB originally PAS-positive and AB-negative reacting mucosubstances become AB-positive. This could be caused by periodic acid oxidation followed by addition of hydrogen sulfite to aldehyde group thus providing secondary basophilic resp. AB positive material.