Blood smears can be fun, but be safe and sterile! -K
Assemble everything required whether only the most basic slide is being made or a more complex preparation. It’s always a good idea to begin with the required materials and a clean work area.
- Use a sterile lancet to procure two drops of blood from ones own, or a volunteers, (alcohol swabbed) finger tip. Discard the first drop and place the second on a very clean slip.
- With a second slip pull a smear on the first slip.
- Dry and fix the smear by ones preferred method. It’s simplest to grip the slide firmly and wave it about until dry.
- Stain the smear with ones preferred stain. For Wright’s stain: a. Drop on stain solution to cover the smear and leave for minimum of two minutes. b. Drop on an equal volume of distilled water and leave in place until a greenish scum forms on the surface (1-4 additional minutes). c. Rinse with a few more drops of distilled water. d. Dry in air, do not blot.
- If desired mount under a coverslip with neutral balsam or green euparal.
- Clean, label, and store slide.
Now for a few notes:
If you have trouble getting a suitable amount of blood try using a larger gauge lancet. A common 33 gauge lancet is very narrow and might not be successful for some people, but a 10 gauge (most common size for Unistik spring loaded lancets) might be too painful for others. A 28 gauge is apt to be more universally acceptable.
When pulling the smear one may find that by holding the second slip at a shallower or steeper angle the thickness of the smear can be controlled in a limited way. The speed at which the smear is made also has some effect on thickness. Try to maintain a consistent speed every time but don’t be afraid to experiment to find the angle that is most successful at ones own speed.
Do not try to fix with any substance (except for some methods of vapor fixation) before the smear has been dried, it will come off, the smear will be ruined. If one is working with a large number of smear it may be better to dry and fix in an oven.
Stains are available in many forms. Even something as common Wright’s stain may be found in a one step buffered solution, as a more traditional un-buffered preparation, or a powder. The directions above will give acceptable results with a buffered or un-buffered solution. When provided, follow the manufacturers directions for the stain used. Additionally, remember that all stains have a shelf life. Most solutions of Wright’s will only work their best after seasoning for a few weeks, and lose their potency after eighteen months.
A stain like Wright’s will bleach out somewhat in an acid mountant. It is therefore advisable to use an ostensibly neutral mountant like euparal or green euparal (which retains stain brilliance better than regular euparal). Alternatively one might keep a few pieces of marble in their balsam bottle to cut down on its acidity.
Depending on the size of the smear one may find a certain need for longer than average coverslips. 22x50mm and 22x35mm covers are widely available but one may also use a smaller cover that only covers a portion of the smear. In any case one should take the time to clean the area not covered after the mountant had cured.
There’s a great deal more to the microscopy of blood and for many people a simple smear can get dull quickly. However, it’s still an important skill and covers many of the various aspects of mounting in a way that can be moved through rapidly in just a few minutes. It’s useful also as a demonstration of the importance of specimen preparation as even an unstained smear will show more than a simple drop of blood under a coverslip. -K