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Used together treatment yeast in urine asacol 400mg with amex, these offer a rational pathophysiologic approach to the laboratory diagnosis of anemia medicine x stanford purchase generic asacol on line. Physiologic Hypoproliferation Excessive M a t u r a t i o n destruction or loss abnormality of red cell Aplastic anemia Hemolytic anemia Megaloblastic anemias Myelophthisic anemia Blood loss M y e l o d y s p l a s i a treatment yeast infection child buy generic asacol on-line, including sideroblastic anemia Renal insufficiency Thalassemia Chronic disease Iron deficiency Endocrine deficiency Stratus 17 medicine 606 buy 400 mg asacol with mastercard. Microcytic anemias An important mechanism of anemia is defective hemoglobin synthesis, which results in small, poorly hemoglobinized erythrocytes. After Wright staining, instead of red cells with pink hemoglobin filling the cytoplasm, the cells are pale with only a rim of 239 Hematology hemoglobin. Since hemoglobin is made up of two components, either of two pathophysiologic mechanisms can lead to decrease hemoglobin synthesis-defective heme or decreased globin production. Heme is made up of iron and porphyrins; deficiencies in either affect heme production. Deficiency of iron store, failure to utilize iron properly, and defective heme or porphyrin synthesis are characteristic of iron deficiency anemia, anemia of chronic disease, and the sideroblastic anemias, respectively. In thalassemia syndromes, globin production is decreased, thereby hindering hemoglobin synthesis and producing a microcytic anemia. Iron deficiency anemia Iron deficiency is the commonest cause of anemia in every country of the world. This is because the body has a limited ability to absorb iron and excess loss of iron due to hemorrhage is frequent. Iron is incorporated form plasma transferrin into developing erythroblasts in the bone marrow and into reticulocytes. Only a small proportion of plasma iron comes from dietary iron absorbed through the duodenum and jejunum. It contains up to 20% of its weight as iron and is not visible by light microscopy. Hemosiderin is an 241 Hematology insoluble protein-iron complex of varying composition containing about 37% of iron by weight. It is probably derived from partial lysosomal digestion of aggregates of ferritin molecules and is visible in macrophages by light microscopy after staining by Perls (Prussian blue) reaction. Iron is also present in muscle as myoglobin and in most cells of the body in ironcontaining enzymes,. This tissue iron is less likely to become depleted than hemosiderin, ferritin and hemoglobin in states of iron deficiency, but some reduction of heme-containing enzyme may occur in severe chronic iron deficiency. Dietary iron Iron is present in food as ferric hydroxides, ferric-protein complexes and heme-protein com complexes. Both the iron content and the proportion of iron absorbed differ from food to food; in general, meat and, in particular, liver is a better source than vegetables, eggs or dairy foods. The average Western diet contains 10-15mg of 242 Hematology iron from which only 5-10% is normally absorbed. The proportion can be increased to 20-30% in iron deficiency or pregnancy but, even in these situations, most dietary iron remains unabsorbed. Iron absorption this occurs through the duodenum and less through the jejunum; it is favored by factors such as acid and reducing agents keeping the iron soluble, particularly maintaining it in the ferrous rather than ferric state. Excess iron is combined with apoferritin to form ferritin, which is shed into the gut lumen when the mucosal cell reaches the tip of the intestinal villus. In iron deficiency, more iron enters the cell and a greater proportion of this intramucosal iron is transported into portal blood; in iron overload, less iron enters the cell and a greater proportion of this is shed back into the gut lumen. Iron transport Most internal iron exchange is concerned with providing iron to the marrow for erythropoiesis. This protein is synthesized in the liver, has a half-life of 8-10 days, and is capable of binding two atoms of iron per molecule. Normally it is one-third saturated but there is a diurnal variation in serum iron, the highest values occurring in the morning and the lowest in the evening. When plasma iron is raised and transferrin is saturated, the amount of iron transferred to parenchymal ells,. Iron requirements the amount of iron required each day to compensate for losses from the body and growth varies with age and sex; it is highest in pregnancy and in adolescent and menstruating females. These groups, therefore, are particularly likely to develop iron deficiency if there is additional iron loss or prolonged reduced intake. Causes Chronic blood loss, especially uterine or from the gastrointestinal tract is the dominant cause. Half a liter of whole blood contains approximately 250mg of iron and, despite the increased absorption of food iron at an early stage of iron deficiency, negative iron balance is usually in chronic blood loss. Increased demands during infancy, adolescence, pregnancy, lactation an in menstruating women account for the prevalence of latent iron deficiency (absent iron stores without anemia) and a consequent high risk of anemia in these particular clinical groups. Newborn infants have a store of iron derived from the breakdown of excess red cells. From 3 to 6 months, there is a tendency for negative iron balance to occur due to growth. It has been estimated to take 8 years for a normal adult male to develop iron deficiency anemia solely due to a poor diet or malabsorption resulting in no iron intake at all. Bone marrow iron • Bone marrow examination is not essential to assess iron stores except in complicated cases, but iron staining is carried out routinely on all bone marrow aspirations that are performed for any reason. Anemia of Chronic disorders One of the most common anemias occurs in patients with a variety of chronic inflammatory and malignant diseases. The anemia is only corrected by successful treatment of the underlying disease and does not respond to iron therapy despite the low serum iron. Sideroblastic anemia this is a refractory anemia with hypochromic cells in the peripheral blood and increased marrow iron; it is defined by the presence of many pathological ring sideroblasts in the bone marrow. These are abnormal erythroblasts containing numerous iron granules arranged in a ring or collar around the nucleus instead of the few randomly distributed iron granules seen when normal erythroblasts are stained for iron. In the hereditary forms, the anemia is characterized by a markedly hypochromic and microcytic blood picture. In the hereditary and primary acquired diseases, 15% or more of marrow erythroblasts are ring sideroblasts. Ring sideroblasts also occur with lesser frequency in the marrow disorders, especially the other types of myelodysplasia, the myeloproliferative diseases, acute myeloid leukemia 252 Hematology and myeloma. They may also occur in the bone marrow of patients taking certain drugs, excess alcohol or with lead poisoning. Lead poisoning Lead inhibits both heme and globin synthesis at a number of points. The anemia may be hypochromic or predominantly hemolytic, and the bone marrow may show ring sideroblasts. Clinically they are divided into hydrops fetalis, fithalassemia major, which is transfusion dependent, thalassemia intermedia characterized by moderate anemia usually with splenomegaly and iron overload, and thalassemia minor, the usually symptomless carrier. As there is duplication of the fi-globin gene, deletion of four genes is needed to completely suppress fi chain synthesis. Since the fi chain is essential in fetal as well as in adult hemoglobin, deletion of both fi genes on both chromosomes leads to failure of fetal hemoglobin synthesis with death in utero (hydrops fetalis. Hemoglobin electrophoresis is normal but occasionally Hb H bodies may be observed in reticulocyte preparations. Beta-thalassemia syndromes Anemia in fi–thalassemia is a result of (1) decreased synthesis of the fi-globin chains of hemoglobin and (2) precipitation and subsequent removal of excess fi-globin chains, which in turn lead to ineffective erythropoiesis and hemolysis. Hypochromia, microcytosis, fragmented forms, and basophilic stippling are found in blood from thalassemia patients. The hypocrhomia is a result of decreased 254 Hematology cellular content of hemoglobin, a major defect in thalassemia. The bone marrow is hyperplastic but the reticulocyte count only moderately increased. The production abnormality is due to ineffective erythropoiesis, that is, destruction of immature erythroid cells in the bone marrow. Several forms of macrocytosis are not accompanied by megaloblastic changes and some of these are relatively common. It is to be distinguished from the swelling of the red cell membrane that accounts for target cell in some patients with obstructive jaundice. Some authors 255 Hematology believe that it is the result of the reticulocytosis that accompanies the hemolytic component of the anemia associated with liver dysfunction. Similarly, macrocytosis, often in the absence of anemia, is seen in patients who consume large amounts of alcohol, and this is sometimes used as a criterion for the diagnosis of chronic alcoholism.
He first learns to build a certain grade of body and afterwards he learns to live in it treatment 12th rib syndrome buy generic asacol 400 mg on-line. All men work unconsciously at the building of their bodies during ante-natal life until they have reached the point where the quintessence of former bodies—which they have saved—is to be built in 7 medications that can cause incontinence purchase asacol pills in toronto. It will therefore be seen that the more a man advances and the more he works on his vehicles treatment zamrud buy asacol 800mg mastercard, thus making them immortal medications journal cheap asacol 400mg on-line, the more power he has to build for a new life. The advanced pupil of an occult school sometimes commences to build for himself as soon as the work during the first three weeks (which belongs exclusively to the mother) has been completed. When the period of unconscious building has passed the man has a chance to exercise his nascent creative power, and the true original creative process— Epigenesis —begins. Thus we see that man learns to build his vehicles in the Heaven World, and to use them in the Physical World. The more we learn of her wonders, the more we realize that this world system is not the vast perpetual motion machine unthinking people would have us believe. It would be quite as logical to think that if we toss a box of loose type into the air the characters will have arranged themselves into the words of a beautiful poem by the time they reach the ground. The greater the complexity of the plan the greater the argumental weight in favor of the theory of an intelligent Divine Author. Here, by the ineffable harmony of this higher world, it is strengthened for its next dip into matter. After a time comes the desire for new experience and the contemplation of a new birth. This conjures up a series of pictures before the vision of the spirit—a panorama of the new life in store for it. After he has chosen and begun his journey it is not sure that he can change to another route during the trip. He may stop over in as many places as he wishes, within his time limit, but he cannot go back. Thus as he proceeds on his journey, he becomes more and more limited by his past choice. If he had chosen a steam road, using soft coal, he must expect to be soiled and dusty. Had he chosen a road burning anthracite or using electricity he would have been cleaner. He may have to live a hard life, but he is free to choose whether he will live it cleanly or wallow in the mire. Other conditions are also within his control, subject to limits of his past choices and acts. The pictures in the panorama of the coming life, of which we have just spoken, begin at the cradle and end at the grave. This is the opposite direction to that in which they travel in the after-death panorama, already explained, which passes before the vision of the spirit immediately following its release from the dense body. The reason for this radical difference in the two panoramas is that in the before-birth panorama the object is to show the reincarnating Ego how certain causes or acts always produce certain effects. Nature, or God, does nothing without a logical reason, and the further we search the more apparent it becomes to us that Nature is a wise mother, always using the best means to accomplish her ends. In the first place, let us realize and engrave it deep upon the tablets of our memory that the purpose of life is not happiness, but experience. Sorrow and pain are our most benevolent teachers, while the joys of life are but fleeting. This seems a stern doctrine and the heart cries out passionately at even the thought that it may possibly be true. Nevertheless, it is true, and upon examination it will be found not such a stern doctrine after all. If we could place our hand upon a hot stove and feel no pain, the hand might be allowed to remain until it and perhaps the arm were burned away, without our knowing anything about it until too late to save them. It is the pain resulting from the contact with the hot stove which makes us snatch our hand away before serious damage is done. If we outrage morality the pangs of conscience bring us pain that will prevent us from repeating the act and if we do not heed the first lesson, nature will give us harder and harder experiences until at last the fact is forced into our consciousness that the way of the transgressor is hard. This is the method by which the occult student should learn, instead of requiring the lash of adversity and pain. The more willing we are to learn in that way, the less we shall feel the stinging thorns of the path of pain and the more quickly shall we gain the path of peace. We cannot stay in the higher worlds and learn there until we have mastered the lessons of earth life. That would be as sensible as to send a child to kindergarten one day and to college the next. The child must return to the kindergarten day after day and spend years in the grammar school and the high school before its study has developed its capacity sufficiently to enable it to understand the lessons taught in college. He must return many times before he can hope to master all the knowledge in the world of sense. No one earth life, however rich in experience, could furnish the knowledge, so nature decrees that he must return to Earth, after intervals of rest, to take up his work where he dropped it, exactly as a child takes up its work in school each day, after the intervening sleep of night. It is no argument against this theory to say that man does not remember his former lives. We do not recollect our labors in learning to write, yet we have acquired a knowledge of the art of writing, which proves that we did learn. Some people do remember their past, however, as a remarkable instance related at the end of the next chapter will show, and is but one among many. Why should a life of happiness in an eternal heaven be the reward for a good lifefi What benefit could come from a good life in a heaven where everybody is already happyfi Surely in a place where everybody is happy and contented there is no need for sympathy, self-sacrifice or wise counsel! No one would need them there; but on Earth there are many who need those very things and such humanitarian and altruistic qualities are of the greatest service to struggling humanity. Therefore the Great Law, which works for Good, brings man back to work again in the world for the benefit of himself and others, with his acquired treasures, instead of letting them go to waste in a heaven where no one needs them. Previous to taking the dip into matter, the threefold spirit is naked, having only the forces of the four seed atoms (which are the nuclei of the threefold body and the sheath of mind. Its descent resembles the putting on of several pairs of gloves of increasing thickness, as previously illustrated. The forces of the mind of the last incarnation are awakened from their latency in the seed atom. This begins to attract to itself materials from the highest subdivision of the Region of Concrete Thought, in a manner similar to that in which a magnet draws to itself iron filings. Its attractive power is of a certain kind and is limited to a certain quantity of that kind. It can take, in each Region, nothing except the material for which it has an affinity and nothing beyond a certain definite quantity even of that. Thus the vehicle built around this nucleus becomes an exact counterpart of the corresponding vehicle of the last incarnation, minus the evil which has been expurgated and plus the quintessence of good which has been incorporated in the seed atom. The material selected by the threefold spirit forms itself into a great bell-shaped figure, open at the bottom and with the seed atom at the top. If we conceive of the illustration spiritually we may compare it to a diving-bell descending into a sea composed of fluids of increasing density. The matter taken into the texture of the bell-shaped body makes it heavier, so that it sinks into the next lower subdivision and takes from that its proper quota of matter. Thus it becomes still heavier and sinks yet deeper until it has passed through the four subdivisions of the Region of Concrete Thought and the sheath of the new mind of the man is complete. It places itself at the top of the bell, inside, and the materials of the seventh Region of the Desire World draw around it until it sinks to the sixth Region, getting more material there, and this process continues until the first Region of the Desire World is reached. The bell has now two layers—the sheath of mind outside and the new desire body inside. The material, of a given quantity and quality, is attracted in the same manner and under the operation of the same law as in the case of the higher bodies, but the building of the new body and the placement in the proper environment is done by four great Beings of immeasurable wisdom, which are the Recording Angels, the Lords of Destiny. It (the vital body) is built by the inhabitants of the Heaven World and the elemental spirits in such a manner as to form a particular type of brain. But mark this, the reincarnating Ego itself incorporates therein the quintessence of its former vital bodies and in addition to this also does a little original work. This is done that in the coming life there may be some room for original and individual expression, not predetermined by past action.
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Those patients with cervical muscle pain demonstrated a significantly higher level of psychoticism compared with the patients without cervical muscle pain and the controls and a significantly higher state anxiety level than the controls treatment for hemorrhoids effective 800 mg asacol. They also demonstrated 41 higher pain intensities in the masseter and temporalis muscles compared with patients without cervical muscle pain medicine and health buy asacol 400 mg fast delivery, however as there were only 10 patients with cervical muscle pain treatment using drugs is called purchase generic asacol pills, these results should be treated with some caution symptoms 7dp3dt buy asacol with a visa. Their main conclusions were that subjects with psychological distress are prone to temporomandibular disorders, or psychological distress is a manifestation of existing chronic pain conditions. Somatisation: Somatisation is defined as fia tendency to experience and communicate somatic distress in response to psychosocial stress and to seek medical help for itfi (Lipowski, 1988. Somatoform disorders are conditions in which the patient reports somatic complaints, yet no physical evidence of organic disease is present (American Psychiatric Association, 1994. Somatisation has been correlated with frequent use of health services (Jyvasjarvi, 2001) and about 20 % of frequent attendees have been classified as chronically somatising patients (Karlsson et al. It has been suggested that patients with masticatory muscle pain may be more prone to report symptoms as compared with normal controls (Wilson et al. Depression: Depression is a disorder that can be defined as a collection of symptoms such as depressed mood, loss of interest or pleasure, weight loss or gain, insomnia or hypersomnia, feelings of worthlessness, and a diminished ability to concentrate (American Psychiatric Association, 1994. Epidemiological studies have shown that depression is the most common mental disorder, with the prevalence of a clinically significant depressive disorder around 4% among males and 8% among females. It affects at least 20% of women and 10% of men during their lifetimes (Kessler et al. There is 42 evidence of a greater prevalence of depressive symptoms in subjects with chronic pain than in controls (VonKorff et al. To avoid causing or exacerbating an existing problem, dentists should not apply too much pressure on the jaw, push the mandible posteriorly, or leave restorations fihighfi. Some patients experience their first symptoms after root canal treatment, whilst other patients may suffer muscle spasms after extraction of third molars. General Summary: the lack of real explanation for temporomandibular joint disorders has set the stage for current aetiological theories. Many professionals utilise treatments which are based on what they perceive to be the cause, as well as their belief in certain treatments. Thus a 43 proportion of patients present with iatrogenic disturbance because of inappropriate surgery, unnecessary occlusal equilibration, unwarranted restorations, orthodontics and incorrect splint therapy (Perry, 1991. It is imperative that aetiological studies from the molecular, biomechanical, neuro-endocrine, physiological, and clinical perspectives be carried out. The participants included all patients between the ages of twelve and nineteen who attended public dental clinics during 2000. The prevalence increased with age and a significant difference was seen between boys (2. They defined a sign as a clinical finding recorded by the examiner of which the patient may have been unaware and a symptom was classed as a finding of which the patient was aware,. She found that, despite methodological and population differences, many consistencies were apparent in the epidemiologic literature. It is also primarily a condition of young and middle-aged adults, rather than of children or the elderly, and is approximately twice as common in females as in males. It does however appear that females outnumber males in seeking treatment and there is wide speculation as to why this is so. Some studies have said that women utilise the health care system more than men, others state that women have a lower tolerance to pain or are more willing to admit to pain than men (Gray et al. Recent research has focused attention on the relationship between sex hormones and pain. Oestrogen administration may increase the incidence of chronic pain (Dao and LeResche, 2000) and this can be explained by actions brought about at a central and peripheral level. In addition it has been found that the pain perception in women varies across the menstrual cycle, with temporomandibular pain at its highest in the pre-menstrual period and during menses (LeResche et al. At the second assessment the results showed that 66% of subjects in both age groups had clinical signs, while 62 % of the 11 year olds and 66 % of the 15 year olds reported subjective symptoms. In most cases the signs were mild, but 11% of the 11 year olds and 17% of the 15 year olds had moderate or, in a few cases severe, signs of dysfunction. Most of the children with subjective symptoms had occasional symptoms but 3% in the younger and 11% in the older age group had frequent symptoms. When comparisons were made with the findings four years earlier at the first assessment, it was noted that the subjective symptoms were greater in frequency in the younger children and the clinical signs had increased in both groups. Nine hundred and twenty Swedish subjects were examined and the questions and clinical examination parameters were in accordance with those suggested by Helkimo in 1974. They found that reported symptoms decreased with age, whilst clinical signs increased. The Indices do not contain several key operational definitions, such as muscle and joint palpation pressures, nor does it endorse a method for scoring joint sound severity. Other problems associated with the Indices relate to the validity and reproducibility (van der Weele and Dibbets, 1987. The instrument was designed to have clearly defined objective criteria, simple clinical methods, and ease in scoring. It is divided into the Dysfunction Index and the Palpation Index and the reliability of both indices was evaluated in the original study (Fricton and Schiffman, 1986. This resulting score does not fiintuitivelyfi describe the patient however (Clark et al. The subjective nature of some items demands that the same rater, who is unaware of the management status of the patient, perform both evaluations. If multiple raters are used, it is recommended that the raters discuss all items and compare scoring of fidemonstration subjectsfi before the study and use a pressure algometer for muscle palpation. The myogenous classification is often further subdivided into muscular hyperarousal due to stress and muscular abnormality associated with parafunctional oral habits (e. The arthrogenous category is subdivided on the basis of specific structural abnormalities (e. In addition these classifications are not always clear, and there can be a considerable overlap or progression from one category to the other (Kuttila et al. Excellent reliability was found for the vertical range of motion measures and for summary indices measuring the overall presence of a clinical sign that could arise from several sources (for example, summary indices of muscle palpation pain. These modest reliabilities could arise from examiner error because clinical signs themselves are unreliable and change spontaneously over time, thus making it difficult to find the same sign on successive examinations. They also found that, without calibration, experienced clinicians showed low reliability with other clinicians. It also includes assessment of behavioural, psychological and psychosocial factors. The diagnostic agreement was determined for the absence of internal derangement, disc displacement with reduction, disc displacement without reduction, and osteoarthrosis. Use of the Kappa statistic test indicated a poor diagnostic agreement between the clinician and the radiologist. This is based mainly on the description of arch form, tooth position and tooth contacts in the intercuspal position. As such, considerable emphasis is placed on this static occlusal relationship (Clark and Evans, 1998. It has been assumed that an ideal static occlusion is synonymous with an ideal functional occlusion (Andrews, 1976; Roth, 1976), however this may not necessarily be the case. Thus it is important to evaluate the features that are thought to be harmonious with an ideal functional occlusion and those which may be detrimental. If epidemiological studies fail to find this occlusal relationship in the natural dentition, the question arises why should this be the goal following orthodontic treatmentfi Whilst every effort should be made to achieve this goal, treatment need not be unduly lengthened in order to do so. Posterior relationships during lateral excursions Posterior lateral excursions can be either group function or canine guided. Group function occurs when the buccal cusps of the posterior teeth on the working side are in contact during the entire lateral movement and there is no tooth contact on the nonworking side. Canine guidance is said to occur during lateral excursion, when contact occurs between the upper and lower canine and the first premolar on the working side only. It is based on the concept that the canine is the most suitable tooth to guide mandibular excursions for the following reasons: 1.
Preparation of blood films on glass slides has the following advantages: • Slides are not easily broken • Slides are easier to label • When large numbers of films are to be dealt with medications 10325 discount asacol 800 mg otc, slides will be found much easier to handle inoar hair treatment asacol 400mg overnight delivery. Wedge method (Two-slide method) • A small drop of blood is placed in the center line of a slide about 1-2cm from one end medicine 852 purchase asacol. Another slide medicine lock box order asacol australia, the spreading slide placed in front of the drop of blood at an angle of 300 to the slide and then is moved back to make contact with the drop. The drop will spread out quickly along the line of contact of the spreader with the slide. It is essential that the slide used as a spreader have a smooth edge and should be narrower in breadth than the slide on which the film is prepared so that the edges of the film can be readily examined. If the edges of the spreader are rough, films with ragged tails will result and gross qualitative irregularity in the distribution of cells will be the rule. The bigger leucocytes (neutrophils and monocytes) will accumulate in the margins and tail while lymphocytes will predominate in the body of the film. If these are not available, writing can be done by scratching with the edge of a slide. If the drop is not too large and if the cover glasses are perfectly clean, the blood will spread out evenly and quickly in a thin layer between the two surfaces. After they are stained they are mounted film side down with permount film side down on glass slides. Spinner method 70 Hematology Blood films that combine the advantages of easy handling of the wedge slide and uniform distribution of cells of the coverglass preparation may be made with special types of centrifuges known as spinners. The spinner slide produces a uniform blood film, in which all cells are separated (a monolayer) and randomly distributed. White cells can be easily identified at any spot in the film On a wedge smear there is a disproportion of monocytes at the tip of the feather edge, of neutrophils just in from the feather edge, and of both at the later edges of the film. This is of little practical significance, but it does result in slightly lower monocyte counts in wedge films. Desirable qualities of a thin blood film • the availability of sufficient working area. Preparation of thick blood smears Thick blood smears are widely used in the diagnosis of blood parasites particularly malaria. It gives a higher percentage of positive diagnosis in much less time since it has ten times the thickness of normal smears. Five minutes spent in examining a thick blood film is equivalent to one hour spent in traversing the whole length of a thin blood film. Method Place a small drop of blood on a clean slide and spread it with an applicator stick or the corner of another slide until small prints are just visible through the blood smear. Which technique of blood film preparation is commonly employed and how is the method of preparationfi What are the possible effects of using a blood sample that has been standing at room temperature for some time on blood cell morphologyfi Jenner (1880) found that the precipitate formed when eosin and methylene blue are mixed could 74 Hematology be dissolved in methyl alcohol to form a useful stain combining certain properties of both parent dye stuffs. Romanowsky (1890) found that when old (ripened and therefore "polychromed") methylene blue solution is mixed with eosin and the precipitate dissolved in methyl alcohol, a stain results that has a wider range than Jenners stain staining cell nuclei and platelet granules (which Jenners mixture failed to stain. Principle of staining Acidic dyes such as eosin unites with the basic components of the cell (cytoplasm) and hence the cytoplasm is said to be eosinophilic (acidic. Conversely, basic stains like methylene blue are attracted to and combine with the acidic parts of the cell (nucleic acid and nucleoproteins of the nucleus) and hence these structures are called basophilic. Romanowsky stains in common use 75 Hematology Modern Romanowsky stains in common,. Wright stain In its preparation, the methylene blue is polychromed by heating with sodium carbonate. Place the air-dried smear film side up on a staining rack (two parallel glass rods kept 5cm apart. When it is planned to use an aqueous or diluted stain, the air dried smear must first be fixed by flooding for 3-5 minutes with absolute methanol. Dilute with distilled water (approximately equal volume) until a metallic scum appears. Without disturbing the slide, flood with distilled water and wash until the thinner parts of the film are pinkish red. Leishman Stain In its preparation, the methylene blue is polychromed by heating a 1 % solution with 0. Staining method the method is similar to that used in Wrights stain except for step 3. With Leshmans stain, dilution is effected with approximately two volume of distilled water to one volume of stain (the best guide is the appearance of a metallic scum. Giemsa stain Instead of empirically polychromed dyes, this stain employs various azure compounds (thionine and its methyl derivative) with eosin and methylene blue. It is commonly used in combination with Jenner or May – Grunwald stains it constitutes panoptic staining". Staining of thick smears the stains used employ the principle of destroying the red cells and staining leucocytes and parasites. Cover the air-dried smear with a 1:10 diluted Giemsa using buffered distilled water at pH 6. Panoptic staining Panoptic staining consists of a combination of a Romanowsky stain with another stain,. This improves the staining of cytoplasmic granules and other bodies like nucleoli of blast cells. Dry the films in the air then fix by immersing in a jar containing methanol for 10-20 minutes. Transfer the slides without washing to a jar containing Giemsa stain freshly diluted with 9 volumes of buffered water pH 6. Dry the films in the air then fix by immersing in a jar containing methanol for 10-20 minutes. It this water-based Romanowsky stain is composed of two solutions, Fields stain A and Fields stand B. It is buffered to the correct pH and neither solution requires dilution when staining thick films. The rapid technique is ideally suited for staining blood films from waiting outpatients and when reports are required urgently. Thin film Fields staining technique Required Fields stain A Fields stain B, diluted 1 in 5 Buffered pH 7. Place the slide on a staining rack and cover the methanol-fixed thin film with approximately 0. Add immediately an equal volume of Fields stain A and mix with the diluted Fields stain B. The stain can be easily applied and mixed on the slide by using 1ml graduated plastic bulb pipettes. Wipe the back of the slide clean and place it in a draining rack for the film to air-dry. Thick film Fields staining technique Required Container of fields stain A Container of Fields stain B Two containers of clean water (need not be buffered) Method 1. Holding the slide with the dried thick film facing downwards, dip the slide into Fields stain A for 5 seconds. Drain off the excess stain by touching a corner of the slide against the side of the container. Wipe the back of the slide clean and place it upright in a draining rack for the film to air-dry. Excessively blue stain • Causes: too thick films, prolonged staining, inadequate washing, too high alkalinity of stain or diluent • Appearance: erythrocytes-blue green, nuclear chromatin-deep blue to black, granules of neutrophils-deeply stained and appear large and prominent. Excessively pink stain • Causes: insufficient staining, prolonged washing, too high acidity of the stain or buffer (exposure of stain or buffer to acid fumes. Precipitate on the film • Causes: unclean slides, drying during the period of staining, inadequate washing of slide at the end of the staining period • Correction: use clean slides, cover the smear with generous amount of the stain, wash the slide until thinner parts of the film are pinkish 84 Hematology Review Questions 1. Describe the appearance of cells and cell components in Romanowskystained thin blood films. Introduction Visual counting of blood cells is an acceptable 86 Hematology alternative to electronic counting for white cell and platelet counts. It is not recommended for routine red cell counts because the number of cells which can be counted within a reasonable time in the routine laboratory will be too few to ensure a precise result. Yet it is still necessary for the technologist to be able to use this method effectively and to know its limitations.