Dry Smears Research Articles

Wet smear method by a water sprayer and temporary decontamination method by a vacuum cleaner.

Improving the smear efficiency and the stability of smear method on the monitoring of radioactive floor surface contamination, wet smear method by a water sprayer was designed and examined. And as the temporary decontamination method applied to the wide spread radioactive floor surface contamination, the method by a vacuum cleaner (vacuum cleaner method) was examined. These examinations were carried out by the experiment using the lonleum floor sheets waxed and the various types of pharmaceutical compounds labeled with 99mTc. On wet smear method, the smear efficiencies were larger than 50% and coefficient of variations by far smaller than that of dry smear method (usual smear method) : for all the radiopharmaceuticals examined. These results showed the effectiveness of wet smear method. On vacuum cleaner method, the decontamination coefficients were comparatively large and almost same to that by wet duster method (decontamination method by using the wet duster) except Ringer's solution and DMSA what had less removable part of contamination. These results showed the effectiveness of vacuum cleaner method for the removable parts of contamination.

Some chromosomal information in Haemonchus contortus Cobb, 1898.

The present investigation have been carried out on the chromosomes of Haemonchus contortus by heat dry smear technique. The diploid chromosome number in the male of this species is 11 while in the female it is 12. The sex mechanism is of XO/XX type. Heteropycnotic sex chromatin body has been observed at the periphery of interphase nucleus of the male. Meiosis shows that the male and the female both are chiasmatic. Sex chromosome X is easily distinguishable in the male sex due to the differential staining behaviour and an univalent nature.

Characteristics of the Membrane Defect in the Hereditary Stomatocytosis Syndrome

Abstract Cation permeability and lipid composition have been studied in the red cells of five patients with various features of the hereditary stomatocytosis syndrome. Hemolysis was compensated in four patients, and only one patient was anemic. Cell NA+ was increased an average of 3 mueq per ml cells and cell K+ decreased 14 mueq per ml cells. Both active and passive fluxes of Na+ and K+ were increased by two to six times normal. Tritiated ouabain binding was increased an average of 2.5- fold, suggesting a proportionally greater number of cation pumps per cell. The coupling ratio of active Na+:K+ fluxes was normal (3:2). Calcium permeability was increased compatible with the degree of reticulocytosis, and cell Ca2+ content was normal. The lowered sum of Na+ plus K+ was associated with a high MCHC and low cell water. When examined in wet preparations, red cells assumed either a bowl-shaped or an irregular contour, and they appeared as target cells on dry smears. Only when cell water was increased in hypotonic media were stomatocytes seen on smear. The total lipid content of red cells was increased in four patients, although it was normal in one. The mole ratio of cholesterol to phospholipid was always normal; however, phospholipid analysis showed an increased proportion of phosphatidyl choline. The abnormal cells were osmotically resistant due to both an increased membrane surface area and a low total cation content. These patients show two hallmarks of hereditary stomatocytosis: bowlshaped red cells observed on wet preparations and a marked increase in Na+ and K+ permeability. The heterogeneity of this syndrome in our patients and in others reported with hereditary stomatocytosis appears to result from (1) variability in the increase in surface area which results from an excess of membrane lipid content, particularly phosphatidylcholine, and (2) a variability in cell water content which may be either decreased or increased as a result of changes in the sum of Na+ plus K+ ions.

Characteristics of the membrane defect in the hereditary stomatocytosis syndrome

Cation permeability and lipid composition have been studied in the red cells of five patients with various features of the hereditary stomatocytosis syndrome. Hemolysis was compensated in four patients, and only one patient was anemic. Cell NA+ was increased an average of 3 mueq per ml cells and cell K+ decreased 14 mueq per ml cells. Both active and passive fluxes of Na+ and K+ were increased by two to six times normal. Tritiated ouabain binding was increased an average of 2.5-fold, suggesting a proportionally greater number of cation pumps per cell. The coupling ratio of active Na+:K+ fluxes was normal (3:2). Calcium permeability was increased compatible with the degree of reticulocytosis, and cell Ca2+ content was normal. The lowered sum of Na+ plus K+ was associated with a high MCHC and low cell water. When examined in wet preparations, red cells assumed either a bowl-shaped or an irregular contour, and they appeared as target cells on dry smears. Only when cell water was increased in hypotonic media were stomatocytes seen on smear. The total lipid content of red cells was increased in four patients, although it was normal in one. The mole ratio of cholesterol to phospholipid was always normal; however, phospholipid analysis showed an increased proportion of phosphatidyl choline. The abnormal cells were osmotically resistant due to both an increased membrane surface area and a low total cation content. These patients show two hallmarks of hereditary stomatocytosis: bowlshaped red cells observed on wet preparations and a marked increase in Na+ and K+ permeability. The heterogeneity of this syndrome in our patients and in others reported with hereditary stomatocytosis appears to result from (1) variability in the increase in surface area which results from an excess of membrane lipid content, particularly phosphatidylcholine, and (2) a variability in cell water content which may be either decreased or increased as a result of changes in the sum of Na+ plus K+ ions.

Cross immunofluorescent reactions between human and rodent malaria.

By the immunofluorescent antibody method, positive serological diagnosis of human malaria was made using Plasmodium falciparum or P. berghei with a titer of 1:4 or higher as an antigen. There was no difference in titer with either species of plasmodium, and no false positive result was obtained among 30 control sera. The test is simple and may be carried out on dry smears that have been stored in a deep freezer. P. berghei berghei shows considerable cross-reactivity with P. falciparum in the serologic method as well as in the localization of parasites in human tissue. It appears to be a useful antigen which can easily be maintained in laboratory mice.

Atlas of Bone-Marrow Pathology.

A reviewer must always ask himself what there is in a new book which warrants its publication; ie, is there something new, original, or is it merely a correction of false old ideas? In the case of this atlas, there is nothing which could not be found in many other atlases, some of which are at least 20 years old. The illustrations and reproduction are of excellent quality but consist entirely of dry smears without the use of even a single section of the bone marrow. There is no attempt at correlation of the clinical course of disease with bone marrow histopathologic findings. The unsophisticated reader might obtain the idea from this atlas that each and every disease has a certain immutable pattern, that there is no progression in the disease, and no interaction between the patient and his environment as the disease progresses. On pages 42 through 44, in

Growth of Trichinella spiralis Larvae from Birth to Day 13 Postinoculation in the Male Albino Rat

Length measurements of Trichinella spiralis were made daily (from 5 to 13 days postinoculation) on larvae recovered from adult females from the small intestine, abdominal cavity fluid, thoracic duct lymph, blood, and the skeletal muscle of male albino rats. Mean length measurements obtained were as follows: newly born larvae (95 u); larvae from abdominal fluid (111 Au); larvae from lymph (105 ,t); larvae from blood (115 /u); and larvae from skeletal muscle on days 5 to 7 postinoculation (106 At). Analysis of the above data using Student's t test indicated significant differences (P 0.10). Using a 2-factor analysis of variance, effects of the two main pathways (blood vs. abdominal fluid), days postinoculation, and individual rats were significant (P < 0.001) influences on larval lengths. The data suggest that larval length is dependent upon the host's and the parasite's physiology. Explanations are given to account for the observed length differences. The life cycle of Trichinella spiralis (Owen, 1835) Railliet, 1895, is well established; nevertheless, certain discrepancies still persist. examining the literature, conflicting size measurements of newly born larvae, migratory larvae, and larvae recovered from skeletal muscle at the time of their first appearance became evident (Table I). Some workers (Roth, 1939; Pambuccian and Cironeanu, 1962; and others) give length measurements indicating that larvae released by females and those recovered from skeletal muscle at the time of their first appearance are similar in size. Ali Khan (1966) reported that the migratory larvae are 111 /, in length in fixed dry smears and 124 ju in length when heat-killed in saline. He also states that After 148 hr, larvae recovered from muscles were significantly shorter than the blood forms .. but offers no explanation to account for this observation. Other workers, such as Thomas (1965), give length measurements indicating differences between the size of newly born larvae, migratory larvae, and larvae recovered from skeletal muscle at the time of their first appearance. This report is therefore concerned with the growth of T. spiralis larvae from birth to day 13 postinoculation (PI) in the male albino rat. Since nematodes show a more pronounced inReceived for publication 7 April 1970. * Present address: Department of Biological Sciences, Eastern Kentucky University, Richmond, Kentucky 40475. cr ase in length while growing than they do in width (Lee, 1965), comparisons were made only on larval lengths vs. recovery sites. MATERIALS AND METHODS Host and parasite Male albino rats (Sprague-Dawley, Holtzman Co.) were used throughout. They were maintained individually in polycarbonate cages; Purina Laboratory Chow and fresh water were provided ad lib. The strain of T. spiralis used was one that has been maintained in this laboratory since 1956 by passage in albino rats. Infective larvae were recovered by pepsin-HCl digestion of skeletal muscles from stock rats infected at least 60 days previously. Experimental rats weighing 150 to 170 g (x = 159 ? 11) and 42 ? 2 days of age were infected by gavage with 3,000 100 excysted larvae. Recovery of larvae From females: In order to obtain uniform measurements at the time of larviposition, a standard procedure was utilized. On each day PI (from 5 to 13) the rat's entire small intestine was removed, slit lengthwise, and the contents scraped into a triple layer cheesecloth bag (24 by 20 mesh). The intestine was then cut into small segments and these added to the bag. The bag was suspended in a 250-ml beaker containing 0.85% saline. The bottom of each beaker was maintained at 37 C via a slide warming tray, thus creating a thermal gradient. The worms, being positively thermotropic, settled and/or moved to the bottom of the beaker and adult females continued to larviposit. The newly born larvae were then collected by pipette, concentrated by pooling, and suspended in nutrient broth (Difco) to which gelatin was added in order to slow down their movements so that accurate measurements could be made. Length measurements were made immediately thereafter

Rhodanile blue: a rapid and selective stain for Heinz bodies.

Equal volumes of heparinized or EDTA-treated blood and a 0.5% solution of rhodanile blue (E. Gurr, Michrome No. 1156) in 1% NaCl were mixed and allowed to stand for 2 rain. Thin smears were then prepared, air dried and examined under oil. Heinz bodies stained deep purple and contrasted well with the yellow-orange to blue-green cytoplasm. Durable mounts could be made by applying a cover glass with a resinous mediiun to the dry smear (D. P. X. was used). The reticular material in reticulocytes did not stain in 2 min but could be stained by allowing the stain to act 5 min.

The Use of the Supravital Method in a Study of Cells Susceptible to Damage in Smears of Normal and Regenerating Bone Marrow

Abstract Cells susceptible to damage during smearing were identified in the different cellular compartments of normal and regenerating bone marrow of the guinea pig by comparing differential counts on fixed dry smears and on supravitally stained wet preparations. The comparisons are considered valid since 1) the number of damaged cells was negligibly low in supravital preparations; 2) it could be shown that cell classification and 3) cell distribution corresponded on the two types of preparations. In smears of normal marrow the majority of damaged cells were myeloid cells, although the most fragile cell appeared to be the monocyte. In normal marrow, erythroblasts and lymphoid cells did not damage in significant numbers. During recovery from sublethal irradiation the number of damaged cells increased significantly in each cell compartment at the time when that compartment commenced its regeneration. In contrast to normal marrow, at the time of lymphoid regeneration 60 per cent of transitional cells had damaged and at the start of erythroid and myeloid regeneration 40-50 per cent of erythroid and myeloid cells were concealed on smears by damage. Labeling with 3H-thymidine indicated that in smears of normal as well as of regenerating marrow, a large proportion of cells susceptible to damage in the various cell compartments was capable of proliferation.

Chromosome number in the lizard genus Uta (family Iguanidae)

A chromosome number of 34 (12 macro- and 22 microchromosomes) was found to be characteristic of the bone marrow in 47 animals including males from the species Uta antiquus, and both males and females from the following species and subspecies: Uta stansburiana stansburiana, Uta stansburiana stejnegeri, Uta stansburiana elegans, Uta stansburiana klauberi, Uta stansburiana mannophorus, Uta nolascensis, Uta palmeri, and Uta squamata. — Diploid chromosome numbers of 34 and haploid numbers of 17 were found in the nine testis smears examined. — The presence of a large number of hypodiploid figures in the bone marrow smears is attributed to cell fragmentation and the problem of distinguishing the small microchromosomes. — Series of polyploid figures whose chromosome numbers increased in arithmetic rather than geometric progressions were observed in the testis dry smears. Possible alternatives for the origin of these figures are presented. — Problems encountered in the use of chromosome number as a taxonomic character are discussed.

Cervical Dry Smears during the luteal phase and the application of 19 nor-methyl-testosterone in the dairy cow

正常牛,繁殖障害牛,低受胎牛計173頭について黄体機能の検討を目的に,黄体期におけるCDS延276例を調べた。また低受胎牛10頭に対して19 N.M.T.の応用を試み,次の成績を得た。(1) 発情終了後, CDSの結晶像の消失は通常,黄体期の第6~7日頃から認められ,第16日頃から次回発情期にかけて再び結晶形成現象の出現がみられた。(2) 3牛群を通じて,黄体初期から開花期にかけて,発育卵胞が黄体と共存している例がかなりみられた。而して,個体によつては黄体機能よりもむしろ共存卵胞の機能が優位の状態でCDSに反映しているとみられるものがあつた。(3) 低受胎牛群の中には,他の2牛群に比べ,黄体初期から開花期におけるCDSの結晶像の消失が遅延するものが多かつた。(4) 3牛群の黄体の大きさ,発育程度に著しい差は認められなかつた。(5) CDSの所見から推定した開花期黄体機能の低下は個体の泌乳期を通じて泌乳最盛期に起り易い現象ではないかと思われる。(6) CDSの所見を示標にして受胎促進のため,低受胎牛10頭に対し19 N.M.T. 40~130mgを人工授精後,黄体初期に1~3回にわたつて経口投与し,中7頭が本処置を契機に受胎した。(7) 以上を通じて,低受胎牛の中のProgesterone適応症の診断にはCDSの所見が不可欠と思われる。また,このことによつて黄体期における内因性Estrogen濃度に対する外因性Progesterone量の盲目的過剰投与が防止出来ると思われる。

Demonstration of a hyaluronidase-sensitive substance in the perilymph of the cod.

It has been demonstrated that the perilymph surrounding the superior part of the labyrinth of the cod contains an acid mucopolysaccharide. There is every probability that this polysaccharide is of the hyaluronic acid type. We arrived at this conclusion by the following reasons:The isolated substance was of a high molecular weight, and its aqueous solution was highly viscous. Dry smears stained heavily metachromatic with toluidine blue at pH = 6.5; the metachromatic reaction was negative after treatment with streptococcal hyaluronidase and/or with pyocyanine. Judged from viscosimetric investigations the substance is readily disintegrated by the streptococcal enzyme. The properties of flow elasticity and displacement potentials faded away after enzymic digestion.

Histopathologic effects of x-rays, radiophosphorus, nitrogen mustard, urethane, and steroids upon the spleen in leukemias and lymphomas.

Practically all prior investigations of the effect of therapy upon the spleen of patients with leukemias and lymphomas have been limited to studies made at autopsy. It is the purpose of this paper to investigate the histopathologic effects of treatment of these diseases by comparing the morphology of biopsy specimens obtained following treatment with a control biopsy obtained immediately prior to therapy. Materials and Methods Technic: Biopsy of the spleen was performed with a Vim-Silverman needle by transthoracic or transabdominal route (1). A piece of tissue about half to two-thirds as thick as the lead in a pencil (1 mm.) and varying from 1 to 3 cm. in length (usually 2 to 3 cm.) was obtained. The larger spleens were biopsied transabdominally, since this is probably safer in the thrombopenic patient, a category into which most of the cases fell. There were no serious complications except for a temporary fall in blood pressure in a single instance. Splenic tissue was prepared by the Maximow technic: Immediate Zenkerformol fixation, sectioning at 6 micra in nitrocellulose, and staining by hematoxylin eosin azure II (2). Slides were also stained for iron (3), and for connective-tissue fibers by the Mallory-azan technic (4). A small piece of tissue was removed from each biopsy specimen for preparation of dry smears by the abklatch technic (5, 6), in which a piece of spleen is gently touched between two slides. These slides were colorized by Wright's stain. All specimens were examined as unknowns and an objective description was made of the microscopic findings. The latter served as a basis for the conclusions drawn in this study. Smears of splenic tissue were not only useless but often misleading, the most important reason being errors inherent in this technic. A multilayered mass of cells with twisted fibers was found wherever the spleen was touched (abklatch of Downey) or smeared on the slide or cover-slip. Because of distortion or opaqueness, the more immature cells, reticular and stem cells, in this mass could not be studied. As a result, only the more mature cells at the periphery of the splenic “touch” or abklatch were available for study. Mitotic cells and nuclear débris were never seen in the smears. A second reason was that it was impossible to determine which type of splenic tissue (normal white or red pulp, fibrous tissue, or tissue specifically involved in the malignant process) was studied. Consequently it was impossible to be certain whether changes seen in smears of repeat biopsies were due to differences in the type of tissue being sampled or to the effect of treatment. Comparability of Biopsies: A study such as this depends upon the comparability of the tissue obtained in sequential biopsies. In the leukemias the spleen has a sufficiently uniform appearance so that this is not a major problem. Serial biopsies from a lymphomatous spleen, however, may vary.

DÉTECTION MICROSCOPIQUE DE LA FLUORESCENCE PRIMAIRE CHEZ LES BACTÉRIES

Thirty-one bacterial species showed a yellow primary fluorescence (auto-fluorescence) which could be observed visually with a commercial fluorescence microscope and recorded on photographic emulsions.The present work emphasizes the importance of this natural property for the study of bacteria, either for taxonomic purposes or as a preliminary step when using fluorochromes. The lack of an easy and reliable technique for detecting the existence of primary fluorescence in bacteria is probably the reason why very little work has been done in this field. Nothing is quoted about this property of bacteria in most textbooks of bacteriology.We deliberately left over the apparently related subject of the well-known macroscopic fluorescence of some Pseudomonas, Azotomonas, and Azotobacter cultures which is associated with diffusible pigments equally present inside and outside the bacteria. The aim of our study was the detection of the primary fluorescence of the bacterial cell itself.In order to examine and photograph the primary fluorescence of bacteria with greater accuracy, two important improvements in technique were introduced (a) elimination of fluorescent particles, present in all the culture media which we used, by the simple expedient of growing the bacteria on cellophane, (b) increased reliability of black and white microphotographs through the use of the bright-field condenser. By using the old and usual method of fluorescence microscopy with the dark-field condenser and ultraviolet rays, it is not always possible to determine on black and white photomicrographs whether or not a given bacterium is fluorescent. With the latter setup the images can be produced not only by the visible light emitted by the fluorescent bacteria but also by the unfiltered rays which eventually pass through the system of filters during long exposures. To overcome this difficulty, we now use the bright-field condenser with ultraviolet or blue rays and the appropriate selective filters. The specimens are best prepared as dry smears, covered with an Euphos selective filter (cover glass) using no immersion liquid. This combination is more likely to give reliable information on the presence or absence of fluorescence.

Comparison of dry smear technics with wet technics for the cytologic study of urine.

Comparison of dry smear technics with wet technics for the cytologic study of urine.

Absorption Microspectroscopy of Bone Marrow Cells

Abstract Using a microspectrographic technic, reflecting optics and narrow bands of the continuum (5500, 4150 and 2650 Å), the author has obtained pictures of human and rat bone marrow cells in unfixed dry smears as well as in suspensions in vivo; erythroblasts showed different absorption patterns. Violet light microspectrography, specific for heme absorption, allowed us to demonstrate the presence of intranuclear heme granules through various stages of maturation of these cells. This confirmed previous and simultaneous results arrived at by cytochemical and other methods. The presence of nucleic acids has been demonstrated in neutrophil granulations.

Crystal structures in dried smears of gastric mucus.

Several characteristic crystal patterns have been observed microscopically in dry smears of pure gastric mucus from canine Heidenhain pouches. Although their chemical composition has not been established, there is reason to believe that they are neither pepsinogen, pepsin, nor one of the mucins.

NAS-benzidine reaction of blood cells-new simple peroxidase staining method using nickel-ammonium sulfate.

We have devised a simple peroxidase staining method using nickelammonium sulfate, and named this NAS-Benzidine reaction. By means of this we can stain the peroxidase positive granules at a time and more rapidly than the existing methods which use separate solutions to be mixed freshly each time. The NAS-Benzidine solution including benzidine, hydrogen peroxide and nickel-ammonium sulfate (NAS), keeps for six months without significant loss of potency. An additional advantage of this stain is the fact that it may be used not only for dry smears but also for the staining of sections and fresh blood. This paper has already been read before the 56th annual Japanese Anatomical Society, April,1951, and the study was carried out with the grant in aid for foundamental scientific research.

TRANSMISSION OF ANTIANEMIC PRINCIPLE ACROSS THE PLACENTA AND ITS INFLUENCE ON EMBRYONIC ERYTHROPOIESIS: II. Comparison of the Effect of Liver Extract and Pteroylglutamic Acid (PGA)

1. Prehepatic embryonic erythropoiesis is influenced when normal pregnant rats are injected with potent antianemic substances.2. Both liver extract and PGA (folic acid) cause a reduction in mean cell diameters, but the former is slightly more effective.3. The rate of mitosis is increased and the relative proportions of individual phases altered.4. The distribution of light areas in the cytoplasm—negative images of organoids as seen in dry smears—is altered to a greater extent by PGA than by liver extract.5. The in vitro and in vivo action of drugs on erythropoietic cells are discussed.

NUCLEAR STRUCTURE VERSUS NUCLEAR PATTERN

Abstract1. Lymphoid cells from rat and human fetal livers and adult rat lymph nodes were studied by means of various cytologic, cytochemical and optical technics.2. Freshly dried smears preserve cytoplasmic detail with an extraordinary degree of accuracy.3. Mitochondria, both numerous and pleomorphic, are preserved in the cytoplasm which lies beyond the nuclear membrane as well as in the thin layers of cytoplasm above and beneath the nucleus.4. In dry-fixed preparations stained with Romanowsky dyes, mitochondria appear as negative images (light areas) in basophilic cytoplasm. These negative images within the nuclear area contribute to the formation of what hematologists in the past have described as parachromatin.5. The dry smear method of studying lymphoid blood cells produces nuclear patterns formed by the sum total of nuclear and cytoplasmic alterations which reflect functional activity.6. The meaning of the terms nuclear structure and nuclear pattern have been contrasted.