BUCCAL SMEAR:

©David B. Fankhauser, Ph.D.,
Professor of Biology and Chemistry
            University of Cincinnati Clermont College,
Batavia OH 45103
Buccal Smear,
methylene blue
1000x
12 Nov 1986, rvsd 18Oct94, 12Sept95, 16Dec96, 3July97, 17Sept97, 11Sept99, 29June00, 28June01, 6July04
This page has been accessed Counter times since 11 July 2001.
Buccal Smear,
methylene blue
2000x

    The cells which line the inside of your cheeks are classified as a stratified squamous epitheliumtissue and are the surface of a mucous membrane. These flat, scale-like buccal cells (pronounced "buckle") are shed constantly as the tissue is renewed. By gently scraping the inside of your cheek, these cells can be harvested, and when smeared and stained, may be used to illustrate a number of important biological phenomena including cell and tissue structure, oral bacterial floraand morphology, etc. This tissue is non-keratinized and therefore the surfacecells are living and still possess their nuclei, in contrast with shed epidermalcells. See DiFiore's Atlas of Histology, 9th Ed, pp 326&327for similar tissue found lining the vagina. (Note that the keratinized surfacecells of the epidermis have no nucleus.)  Here is a labeled image of a buccal smear stained with methylene blue.

Compare the following steps with those in the Bact. Smear and Staining Protocol .
 

EQUIPMENT
AND SUPPLIES:
 soap and water
microscope slides
dropper bottle of dH2O
dropper bottle of 0.3% methylene blue
toothpicks (optional)
Bunsen burner or alcohol lamp, striker
bibulous paper or paper towel

PREPARATION OF SLIDE AND INDEX FINGERNAIL:
 

1. Clean a microscope slide well withsoap & water, dry with a non-linty paper towel.
Clean under fingernail.
2. Cleanse very thoroughly under thenail of your index finger.
3. Place a small drop of dH 20 in the center of the very clean slide.

HARVEST THE CELLS, PREPARE THE SMEAR:
 

harvest buccal cells
4. GENTLY scrap the inside ofyour cheek to pick up some of the shed stratified squamous cells. Do NOT scrape chunks. A toothpick may be used if you have no fingernails. Gentle scraping is the watchword, there should be no discomfort...
press out the cells from under the nail

apply cells to slide
5. Express the material from under your nail by pressing with your thumb, and press the material into the drop of water on the slide, mix and spread the material around to the size of a dime:


FIX THE SMEAR:
 

6. Pass the slide through the flame several times to fix the smear. Do NOT heat the slide above a temp whichis comfortable.

You are merely "gluing" the smear to the slide.

STAIN THE SMEAR:
 

7. Place a drop of 0.3% methylene blue on the specimen. Let sit for 1 minute.
8. Rinse off the excess stain with tap water. (Do not splash on your white shirt!)
  9. Blot dry with an non-linty paper towel or bibulous paper. Do not rub.
10. Flame again briefly to dry slide.

EXAMINE UNDER THE MICROSCOPE:
 

11. Examine first with the 4x objective, scanning the entire field to find a well-distributed region.  Avoidregions where cells may be piled up to thickly . Then view with the 10x and 40x objectives, illustrating the view at both powers. Note 1) the nucleus, 2) nucleolus, 3) cell boundary and 4) the variety of bacteria colonizing the surface of the cells.
For Microbiology only: you will be instructed on oil immersion use , then illustrate bacterial morphologies with the 100x oil immersion objective.

CLEAN UP:
 

12. When finished, scrub the slide well in hot soapy water, rinse well and drain dry in a plastic test tube holder.