CLINICALBACTERIOLOGY- LABORATORY

WEEK 3
Mr. Ariel E. Salvacion
ASEPTIC TECHNIQUE AND ISOLATION ISOLATION STREAKING
STREAKING
• Open the agar plate insert the loop and make the loop touch
the first sector ONCE, drawing a few inoculum from the first
• PRINCIPLE: sector into the second sector.
- The purpose is to create a region where bacterial cells • Also streak using a zigzag motion but less heavier than the
are far from one another. first sector
• Spreading an inoculum with multiple species can result 3. THIRD SECTOR (Sterilize agin before continuing to 3 rd
in the isolation of individual species with one bacterial sector).
cell creating a colony as it multiplies. • Open the agar plate and insert the loop. Make the cooled
loop touch the second sector once, drawing a few of the
bacteria to the third sector.
• This sector has the widest gap between the rows.
• Once done sterilize the loop and invert the plate so that the
agar is on the up side and incubate the plates for 18-24 hours.

ISOLATION STREAKING PROCEDURE

1. Label your plates (EDGE OF THE PLATE)
• Organism/specimen name • 4 streak isolation method
• date inoculated; and • Same with T streak isolation technique but with an
• plater's name/initials. additional sector. And turning the plate must be in 90
2. Sterilize the inoculating loop by flaming in an alcohol degree angles to give space for each sector.
lamp until red hot. • COMMENTS AND TIPS:
3. Open the culture and collect a sample of the specimen 1. Use the 4 quadrant streak pattern when large amounts of
using the sterile loop bacteria are expected.
4. Remove and hold the test tube cap using your right 2. Always wait until the loop turns glowing red when
hand's little finger sterilizing
5. Flame the tube's mouth using your left hand then insert 3. PRACTICE. Get familiar with your technique to avoid
the loop into the suspension. digging or pitting the agar.
6. Remove a loopful of broth then flame it again over the
test tube's mouth
7. Replace the caps and put the test tube back into the test
tube rack
8. STREAK THE PLATE- T Streak isolation method & 4
streak isolation method

STREAKING OF PLATE
• T streak isolation method
1. FIRST SECTOR:
• Raise the agar plate to insert the loop and make the loop touch
the first sector of the agar.
• Spread the inoculum in the first sector by moving the loop in
a zigzag pattern.
2. SECOND SECTOR (Sterilize the loop again first befor
continuing to 2nd sector)
• Remove the loop from the agar plate and sterilize the loop
before continuing to the second sector.
• Allow the loop to cool off before proceeding to the streaking
process.

MONTALBAN, KIMBERLY N. 1