A well prepared blood smear is necessary for microscopic examination of blood. Increasing the angle makes a shorter, decreasing the angle a longer smear. Do not make the smear too thick or it will fall off the slide. The remarkable property of the romanowsky dyes of making subtle distinctions in shades of staining, and of staining granules differentially, depends on two components. The blood can be obtained from finger heal microhematochrit tubes non heparinised for edta or heparinised for capillary blood wooden stick or glass capillary pipette. It is also used for staining bone marrow aspirates, urine samples and to demonstrate malarial parasites in blood smears. Operate this stain far from an open source of fire. Blood smear preparation and staining cont, 3 3 in a smooth move, draw the spreader slide away it should form a bullet shape. A blood smear is a blood test used to look for abnormalities in blood cells. Slides are just automatically dipped in the stain in the buffer and a series of rinses. The thin blood films are prepared from whole blood with clean, dry and wrapped microscope slides cfr.
Right blood as seen at 400x under light microscope. Learn vocabulary, terms, and more with flashcards, games, and other study tools. Im staining fish blood smears and searching for micronuclei. C m ac m a a drop of blood is placed on a clean microscope slide. A bss is usually performed to verify the automated platelet count, particularly if it is flagged by the analyzer for verification or if it is significantly lower than the lowest limit of the reference range. A peripheral blood smear, or blood film, is a microscope slide made from a drop of blood for microscopic examination. The extra time and care taken during the field season will be rewarded later when the smears must be scanned, and parasites identified and counted. After staining with giemsa, the slide can be examined under the microscope. Making and staining a bloo d smear a wellmade blood smear is a beauty to behold, and likely to yield interesting and significant information for a research project. Preparation of thick blood films and giemsa staining. The word gram is always spelled with a capital, referring to hans christian gram, the inventor of gram staining gram staining principles gram staining is used to determine gram status to classify bacteria broadly.
Module common staining technique microbiology 24 notes indeterminate groups as well. The thick blood films are used for microscopic detection of plasmodium parasites, the causal. Blood cell guide for specimens with low hematocrits. Icsh has published a reference staining method for blood films based on purified azure b and eosin y solutions ichs, 2010. Since good quality control smears are not available commercially, they may be prepared from a patients blood and stored for future use in the following manner. Learn blood smear staining with free interactive flashcards. Blood smear preparation and staining mlab 15 hematology blood smear preparation and. The purpose of preparation of blood smear blood film is to study the morphology of rbcs, differential leukocyte count and reticulocyte count. A variety of methods are available for preparing and staining blood smears, the most common of which are discussed in this atlas. Wait until the thin and thick films are completely dry before staining. The blood smear should occupy the central portion of the slide and should not touch the edges.
Choose from 500 different sets of blood smear staining flashcards on quizlet. The three main blood cells that the test focuses on are. Wrights stain is named for james homer wright, who devised the stain in 1902 based on a modification of romanowsky stain. However, the red blood cells took on more stain too, appearing darker. The excess stain is removed with dilute hydrochloric acid solution acid water.
To ensure that proper staining results have been achieved, a positive smear malaria should be included with each new batch of working giemsa stain. Trichrome staining is inadequate for a definite diagnosis because oocysts will appear unstained. Prior to staining, cells must be fixed to the glass slide with acetone free methanol, either alone or in solution with. Take another clean slide, and holding at an angle of about 45 deg, touch the blood with. The decolourising process is then stopped by immersing the slide in running tap water. Scope and application this procedure describes the preparation of thick blood films. A properly prepared blood smear is essential to accurate assessment of cellular morphology. A simple single differential stain of blood smears using. A blood smear is often used as a followup test to abnormal results on a total blood count cbc to evaluate the various types of blood cells. Besides transporting vital components, the blood plays an important role in the immune functions of the body and is vitally important to coagulation ability of blood to clot properly. Immune hemolytic anemia is classified as autoimmune, alloimmune, or druginduced, based on the antigen that stimulates antibody or complementmediated destruction of red blood cells. B staining of blood smear leave it for 12 minutes for fixation of the smear. Color atlas of clinical laboratory medicine first version. Preparation of thick blood films and giemsa staining projectstudy.
When using the regressive staining method, the nucleus is deliberately overstained with a nonacidified haematoxylin. To use and relate the gram stain to the study of bacterial cell morphology, and as an important step in the identification of a bacterial species. A blood smear scan serves to at least a verify the flagged automated hematology results and b determine if a manual differential leukocyte count needs to be performed. As a general rule for judging the quality of a stained blood film, the laboratory must ensure that all cell types in a blood film. The thickness of the spread when pulling the smear is determined by the 1 angle of the spreader slide the greater the angle, the thicker and shorter the smear, 2 size of the blood drop and 3 speed of spreading. Staining blood and bone marrow films romanowsky stains are used universally for routine staining of blood films, and satisfactory results can be obtained. Apply the maygrunwald solution to the dried smear and wait for 35 min. Slides stained 1 week or longer after they are made turn out too blue.
Bloodfilmpreparationand stainingprocedures berendhouwen,md,phd. Microspherocytes on a peripheral smear and a positive direct antiglobulin test are the characteristic findings. Peripheral smear using leishman stain linkedin slideshare. Dried blood smears may be stored at rt for several days before use.
I have seen methods that have only used a 6% giesma stain solution in sorensons buffer nwani et al. Histology laboratory microscope orientation and blood. It might be used to help identify andor keep an eye on numerous conditions that impact blood cell populations. An unstained blood smear is clearly seen by eye on the microscope slide, however, the blood cells are barely visible under the microscope figure 1. Megan brashear, cvt, vts ecc, demonstrates technique in staining a blood smear for microscopic evaluation. Neel, dvm, dacvp clinical associate professor, clinical pathology nc state college of veterinary medicine raleigh, nc, 27607 introduction although tremendous advances have been made in the field of pointofcare hematology analyzers, examination of a well prepared, well stained blood smear remains the. Neel, dvm, dacvp clinical associate professor, clinical pathology nc state college of veterinary medicine raleigh, nc, 27607 introduction although tremendous advances have been made in the field of pointofcare hematology analyzers, examination of a well prepared, well stained blood smear remains the cornerstone of veterinary. Blood films with excessive tails or gritty feathered ends indicate a spreader edge that is rough or dirty, or an accumulation of leukocytes due to either slow spreading or a very high leukocyte count. Blood smears are used to determine leukocyte differentials, to evaluate erythrocyte, platelet and leukocyte morphology, and, if necessary, to estimate platelet and leukocyte counts. The best staining results are obtained on fresh slides because the blood itself acts as a buffer in the staining process. To learn the techniques of gram staining, nigrosin staining and koh test. Making and staining a blood smear a wellmade blood smear is a beauty to behold, and likely to yield interesting and significant information for a research project. It is hoped that this paper will provide an aid to those who wish to employ the blood smear as a diagnostic tool.
Preparation and staining methods for blood and bone marrow. Blood is located in almost every part of the body, because it. Ensure that the newly prepared blood film is completely dried before staining is performed. Practical of clinical hematology islamic university of gaza. Making and staining a blood smear university of vermont. If humidity is high, dry the slide with a slowspeed fan without moisture or heat, or simply wave the blood film in the air. Use of an excessively large drop of blood will result in too thick a smeartoo small a drop of blood will result in too short a smear. Introduction to peripheral blood smear examination. C m ac m a a spreader slide is used to feather the drop of blood. Histology laboratory microscope orientation and blood smear lab for practicing how to use the microscope do not use the blood smear slide it is too boring for the lower mags. Procedure of regressive papanicolaou staining method. Purpose and criteria for blood smear scan, blood smear.
Staining fish blood smears and looking for micronuclei. Objectives to learn the technique of smear preparation. Staining process has begun, no stat slides can be added in the batch. Determine the overall staining quality of the blood smear. If aequous staining procedures will be used fix blood smear 3 min in cold meoh and dry 10 min on the. C m ac m a blood smear preparation materials and supplies sample edta slides fixative, buffer and stain diffquik water 4. Thick blood smear showing trypanosoma species staining the thin and thick blood smear on the same slide with giemsa stain. Aqueous solutions of stains are stable only after 36.
Prepare a thin and thick smear of the specimen on the same slide which can be done by dividing the slides into two parts and then making the thin smear on one side and thick on the other side. It takes about 510 minutes to stain a batch of smears. The mechanism of action of wrights stain is also discussed. The other is pusher or the spreader slide drop of blood 23 mm in diameter. Staining blood smears with dilutions of 10x or 20x in either water, saline, dilute sodium bicarbonate or vitamin c only gave faint blue staining of white blood cells after 3.
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