Blood Smear Preparation PDF

Blood Smear Preparation Lab: 2 Assistant Lecturer: Ahmed Mekki  ‫قال الشاعر أبي العالء املعري ‪:‬‬ ‫ما باهلا قُطعت يف ربع دينار؟‬ ‫يدٌ خبمس مئني عسجدٍ فُديت‬ ‫ونستجري مبوالنا من النار‬ ‫تناقضٌ ما لنا إال السكوت له‬...

Blood Smear Preparation Lab: 2 Assistant Lecturer: Ahmed Mekki  ‫قال الشاعر أبي العالء املعري ‪:‬‬ ‫ما باهلا قُطعت يف ربع دينار؟‬ ‫يدٌ خبمس مئني عسجدٍ فُديت‬ ‫ونستجري مبوالنا من النار‬ ‫تناقضٌ ما لنا إال السكوت له‬ ‫ماذا يعني لك هذا البيت من الشعر ؟؟؟‬  Examination of thin blood films is important in the investigation and management of anemia, infections, and other conditions which produce changes in the appearance of blood cells and differential white cell count. A blood film report can provide rapidly and at low cost, useful information about a patient’s condition. Aim of blood smear Three basic steps to make blood film 1.Preparation of blood smear. 2.Fixation of blood smear. 3.Staining of blood smear. Making blood films  Methods may be used to make blood smears. 1 The cover glass smear. 2 The wedge smear. Blood Smear Preparation Specimen: Peripheral blood smear made from EDTA- anticoagulated blood. Smears should be made within 1 hour of blood collection from EDTA specimens stored at room temperature to avoid distortion of cell morphology. Blood smears can also be made from finger sticking blood directly onto the slide. WEDGE BLOOD SMEAR  Spreaders. Clean slides. Blood capillary tube or micropipette 10 μL. Equipment  Fill a capillary tube three-quarters full with the anticoagulated specimen. Place a drop of blood, about 2 mm in diameter approximately an inch from the frosted area of the slide. Place the slide on a flat surface, and hold the narrow side of the non frosted edge between your left thumb and forefinger. With your right hand, place the smooth clean edge of a second (spreader) slide on the specimen slide, just in front of the blood drop. Procedure  Allow the blood to spread almost to the edges of the slide. Push the spread forward with one light, smooth, and fluid motion. A thin film of blood in the shape of a bullet with a feathered edge will remain on the slide. Label the frosted edge with patient name, ID# and date. Allow the blood film to air-dry completely before staining. (Do not blow to dry. The moisture from your breath will cause RBC artifact). Hold the spreader slide at a 30° angle, and draw it back against the drop of blood The shape of blood film. 1. A good blood film preparation will be thick at the drop end and thin at the opposite end. 2. The blood smear should occupy the central portion of the slide. 3. The blood smear should not touch the edges. except for point of application. 4. Should be margin free. Characteristics of A Good Smear 1) Drop of blood too large or too small 2) Spreader slide pushed across the slide in a jerky manner. 3) Failure to keep the entire edge of the spreader slide against the slide while making the smear. 4) Failure to keep the spreader slide at a 30° angle with the slide. 5) Failure to push the spreader slide completely across the slide. 6) Irregular spread with ridges and long tail: Edge of spreader dirty or chipped; dusty slide 7) Holes in film: Slide contaminated with fat or grease 8) Cellular degenerative changes: delay in fixing, inadequate fixing time or methanol contaminated with water. Common causes of a poor blood smear A: Blood film with jagged tail made from a spreader.. B: Film which is too thick C: Film which is too long, too wide, D: A well-made blood film.  To preserve the morphology of the cells, films must be fixed as soon as possible after they have dried. It is important to preventcontact with water before fixation is complete. Methyl alcohol (methanol) is the choice, although ethyl alcohol ("absolute alcohol") can be used. Methylated spirit (95% ethanol) must not be used as it contains water. Fixation of blood smear Leishman's Thin smear are air dried. Flood the smear with stain. Stain for 1-5 min. Experience will indicate the optimum time. Add an equal amount of buffer solution and mix the stain by blowing an eddy in the fluid. Leave the mixture on the slide for 10-15 min. Wash off by running water directly to the centre of the slide to prevent a residue of precipitated stain. Stand slide on end, and let dry in air. Staining the film