Name: SAMONTE, Dennise Ann R.

Date:
Section: NUR203 Score:

Exercise No. 4
Staining
Bacteria and other microorganisms are usually transparent which makes the study of
morphologic details difficult when they are examined in the natural state. The early methods of
fixing and staining allowed the microbiologist to distinguish many structural features not formerly
seen. The microbiologist is advised to use a young culture for routine staining procedures not
unless specific morphologic feature dependent on the age of the culture is to be demonstrated.

Objectives:
1. The student must be able to know the basic concepts behind staining procedures.
2. The student must be able to prepare smears from stock cultures.
3. The students must be able to describe the morphology of the microorganism after staining.
Materials:
Slide Methylene blue
Wire loop Malachite green
Alcohol lamp Safranin
Glass slides Crystal violet
Microscope
Bacterial suspension
Procedure:
1. Get a clean glass slide and gently heat one side to remove any grease. Slides should be
held along the edges to prevent recontamination with the grease from the fingertips. Lay
the slide on the table with the flamed side up.
2. Sterilize wire loop until it is red hot and allow it to cool.
3. Get 2-3 loopfuls of the bacterial culture (butt, butt slant, slant, plated media) and spread
over an area of 1 inch by ½ inch.
4. Air-dry by laying the prepared smear on the table.
5. Heat fix the smear by passing the slide (with smear side up) over the flame 5x or 6x, then
allow to cool. Fixation of the smear causes the preparation to adhere to the slide and will
not be easily washed off during the staining process.
6. Place the slide on a staining rack and cover the smear with methylene blue or crystal violet
stain. Keep the slides level and do not let it dry.
7. Allow stain to act for 1-2 minutes. Then remove the excess stain by washing gently in tap
water.
8. Blot dry the preparation with absorbent paper. DO NOT RUB.
9. Draw your results.
Butt (Throat swab) Butt slant (Throat swab) Slant (Throat swab)
Plated media A (Throat swab) Plated media B (Throat swab)

Plated media C (Throat swab) Plated media D (Throat swab)

Butt (Skin swab) Butt slant (Skin swab) Slant (Skin swab)

Plated media A (Skin swab) Plated media B (Skin swab)

Plated media C (Skin swab) Plated media D (Skin swab)
Experiment worksheet on Staining
I. Complete the table below
Staining techniques Characteristics Examples
Simple Staining

Differential Staining

Diagnostic antibody
staining

Negative staining

Reference: