BIOL 2070 Study Guide - Midterm Guide: Phase-Contrast Microscopy, Nocodazole, Growth Medium

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13 Aug 2016
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Objective: The purpose of the experiment is to study the effects of the drug Nocodazole on the growth
of Hyphal Tip Growht of Neurospara Crassa.
Protocol
1. Collect the N.crassa that had been incubated for 24 hrs
2. Add 3-4 drops of the growth medium onto one slide (2 cushions) and place a coverslip, this will
serve as the control. Remember to label the slides.
3. Add 3-4 drops of the growth medium mixed with 300 µM of Nocodazole onto another two slides
(4 cushions), this will serve as the experimental. Label the slides.
4. Allow the slides to incubate for 10 minutes.
5. While waiting for the slides to incubate, set up Kohler illumination at 40x magnification in phase
contrast microscopy.
6. Place the slides under the microscope and observe the rate at which the fungus grows.
a. Check the distance the fungus grew every 30 seconds and then once the growth
steadies, record for every minute.
Table 1: Observation table of the hyphal growth of N.crassa for Slide 1 control cushion A.
Time
(Min)
Distance travelled in
ocular units
Distance travelled in
µm
Rate of Growth
(µm/min)
0.5
1
1.5
Table 2: Observation table of the hyphal growth of N.crassa for Slide 1 control cushion B.
Time
(Min)
Distance travelled in
ocular units
Distance travelled in
µm
Rate of Growth
(µm/min)
0.5
1
1.5
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