MICROBIO 4.3

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OBTAINING A PURE CULTURE

Last updated 5:23 AM on 8/12/26
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17 Terms

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Pure culture

Culture growing are only ONE species of cells; originated from ONE cell

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Plating

Process of spreading bacterial culture onto petri dish to isolate colonies

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Supplies for plating

  • Agar plate

  • Culture

  • Sterile loop/swab/needle/wire/L-shaped rod

  • Incubator (37 degrees Celsius)

***Tools & environment MUST be sterilized

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Four-phase dilution

[P1] > [P2] > [P3] > [P4]

***# of phases can inc OR dec depending on culture concentration OR seterile loop can be passed through previous quadrant multiple times

After streaking, samples INVERTED and incubated at 37 degrees C

***Temp & time can be varied to optimize generation time & restrict/enhance certain microbial growth

<p>[P1] &gt; [P2] &gt; [P3] &gt; [P4]</p><p>***# of phases can inc OR dec depending on culture concentration OR seterile loop can be passed through previous quadrant multiple times</p><p>After streaking, samples INVERTED and incubated at 37 degrees C</p><p>***Temp &amp; time can be varied to optimize generation time &amp; restrict/enhance certain microbial growth</p>
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Generation time

Time it takes for bacteria to divide/double

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Quadrant Growth Plate

Tests multiple cultures; will they grow on plate with specified conditions?

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Spread plate

  1. Bacterial culture is serial diluted

  2. Dilute culture is spread on surface of agar by L-shaped agar & placed on turntable

  3. Uniformly spread

Goal: Spread single cells across surface

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Pour plate

  1. Culture serial diluted in broth

  2. When poured onto petri dish, mix culture with warm agar

  3. Mixture & agar solidifies together

Result: Colonies grow THROUGHOUT agar

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Primary advantage of agar plating

Cells are held in place to support formation and visualization of colonies

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Skin specimen collection method

Sterile swab brushed on surface

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Blood specimen collection method

Needle aspiration from vein; anticoagulants included in specimen tube

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Cerbrospinal fluid specimen collection method

Needle aspiration from subarachnoid space of spinal column

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Stomach specimen collection method

Intubation often via nostril

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Urine specimen collection method

Aseptic: catheter is inserted into bladder through urethra

Clean catch: Initial urination washes urethra, THEN specimen is midstream urine

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Lungs specimen collection method

Collection of sputum either by coughing or via catheter

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Diseased tissue specimen collection method

Surgical removal (biopsy)