May 17 Day 4

We coducted a motility test. Positive motility= bacteria spreads from stab
-used asceptic technique to dip a sterile inoculating needle into a broth culture.
-stab the bacteria into the center of the test medium to about halfway or three quarters depth
-incubate at 30'C for 24-48 hours


Bacterial Smear:
1-place a loopful of distilled water in the center of the slide
2-transfer a small amount of culture from the agar surface into the water drop using a sterile loop. Spread the mixture into a thin film
3-Transfer a loopful of liquid culture to the center of the slide using a sterile loop
4-spread the drop of culture into a thin film
5-flame the loop before putting it down
6-allow the smear to air dry
7-pass the slide quickly through flame three times, smear slide up
8-place the slide on a slide warmer(60'C for 10minutes)
9-add 95% methanol to the dried smear for at least 1 minute. Rinse off the methanol
10-Roll the cotton swab in the center of the slide
11-allow the smear to air dry
12-fix the smear by heating

Simple Stain:
1-Cover the fixed smear with several drops of stain
2-rinse the slide with water to remove excess stain
3-blot water from the slide with bibulous paper
4-examine the stained smear under the microscope using oil immersion lens

Gram Stain:
-Place a fixed smear on a rack over a staining tray or sink
-Cover the smear with crystal violet for 20 Seconds
-Rinse the slide with water to remove excess crystal violet
-Cover the smear with Gram's iodine for 1 minute
-Rinse the slide with water to remove excess iodine solution
-decolorize with 95% ethanol or ethanol/acetone. Hold slide at 45' angle while adding decolorizing reagent drop by drop until color stops running
-imediately rinse slide to remove decolorizing agent
-cover smear with safranin for 1 minute
-rinse slide with water to remove excess safranin
-blot water from slide with pieces of bibulous paper

After Examining the smear under the microscope using the oil immersion lens, we concluded our bacteria is Gram Positive