Table of Contents
What is used in preparing a blood smear slide?
Using a microhematocrit tube or small pipet, place a drop of well mixed EDTA anticoagulated blood 3-4 mm in diameter near the frosted end of the slide or 1⁄4 inch from one end of an unfrosted slide. 3. Rest the spreader slide at a 25° angle on the slide to be made.
How do you prepare a blood slide for a microscope?
How to Make a Blood Smear Slide Place a small drop of a liquid sample onto the slide. Take a second clean slide. Hold it at an angle to the first slide. Use the edge of this slide to touch the drop. At this point, either allow the slide to dry so that it can be stained or else place a coverslip on top of the smear.
What are the three methods of preparing a blood smear?
Four different types of smear preparation methods (conventional method, blood film method, drop and rest method, and water-wash method) were carried out according to the standard reference as described below.
What is the most important thing to remember when creating a blood slide smear?
What is the most important thing to remember when creating a blood smear? The important thing to do when creating a blood smear is to create a feather edge .
How can I improve my blood smear?
Maintain a constant smooth motion, angle and even contact. Note – blood is being dragged behind the spreader slide, not in front of the slide. An optimal smear is ¾ the length of the slide and has a feathered edge. Leave the slide to air dry and make more smears if required.
What is blood smear preparation?
Blood films should be made immediately after collection of the blood, because cell morphology deteriorates rapidly after sample collection. EDTA sample or fresh blood immediately from the collection needle, before the contact with anticoagulant can be used.
How routine and thick blood smears are made?
Smears are most commonly made using the “spread” or “wedge” technique. Manual smears are made by placing a drop of blood on one side of a glass slide, and spreading this by rap- idly moving a second glass slide or spreader across the first slide at an angle.
What stain is used for blood smears?
Blood films are routinely stained with a Romanowsky-type stain (e.g., Wright or Wright-Giemsa) either manually or using an automatic slide stainer. Romanowsky-type stains are composed of a mixture of eosin and oxidized methylene blue (azure) dyes.
How do you make a wedge blood smear?
The wedge method: First, place a 2-3 mm drop of blood about 1 cm from the frosted end of a clean slide that is on a flat surface. Second, with the thumb and forefinger of the right hand, hold the end of a second slide or coverglass (“spreader slide”) against the surface of the first slide at an angle of 30-45 degrees.
How does alcohol fix a blood smear?
Blood smears are fixed by dipping in absolute methanol or ethanol for 30 seconds. Fixation is a technique that is varied depending on the quality of results you wish to achieve. No one method is absolute and alterations to time and type of fixation are normal in a biological laboratory.
What is the best anticoagulant for smear preparation?
Ethylene diamine tetra-acetic Acid (EDTA) is the anticoagulant of choice. Samples should be sent to the laboratory as soon as possible.
What kind of edge is needed for a smear?
Look at the slide and grossly check the smear, making sure it has two edges: These two edges include the feathered edge (a thin portion) and the thick edge (a base). Approximately two-thirds to three-fourths of the slide length should be covered by the smear with no irregularities, streaks, or holes.
Why is it important to stain the blood smear?
Examining the monolayer of a well-prepared and properly stained blood smear allows veterinary technicians to assess erythrocyte (RBC), leukocyte (WBC) and platelet morphology. The presence of nucleated red blood cells in peripheral blood of mammals is indicative of bone marrow stress or damage.
How many drops of deionized water should you use to clean the slide?
a. Heat-fix a smear of Escherichia coli as follows: 1. Using the dropper bottle of deionized water found in your staining rack, place 1/2 of a normal sized drop of water on a clean slide by touching the dropper to the slide (Figure 1).
How do you lengthen or shorten a blood smear?
Invert the tube of EDTA blood 10 times. How long should the blood smear extend? It should extend 2/3s of the total slide area.
Why is thick smear not fixed?
Thick smear. It is not fixed in methanol; this allows the red blood cells to be hemolyzed, and leukocytes and any malaria parasites present will be the only detectable elements.
What are the important precautions in preparing a blood smear?
Precautions to be taken during preparation: Angle should be maintained at 45°. Blood drop should be of proper size. Spreader’s edges should be smooth and it should be smaller than the slide on which smear is being made. Pressure applied should be proper. Drop should be pulled with spreader not pushed with it.
How do you make a smear?
Culture from solid media: Using a sterile pipette, add one drop of sterile saline or sterile water to the center of the microscope slide. Aseptically pick a small amount of an isolated colony with a loop and gently mix into the drop of sterile saline or water using circular motions. Mix evenly to make a thin smear.
How do you stain a Giemsa?
Preparation of the Giemsa Stain Stock solution (500ml) Into 250ml of methanol, add 3.8g of Giemsa powder and dissolve. Heat the solution up to ~60 o C. Then, add 250ml of glycerin to the solution, slowly. Filter the solution and leave it to stand for about 1-2 months before use.
