Making Agar Cushion Lab Report
Making Agar Cushions
1. Start with wearing gloves and swabbing them with 70% ethanol to keep the materials that will be used in the experiment sterile.
2. Wipe the work station with Wescodyne and paper towels. It is important to keep the sterile materials such as slides in petri dish, Pasteur pipette in container and forceps covered as much as possible.
3. Place four to five filter papers in the petri dish and drip few drops of water on the strips, this will maintain moisture in the agar cushions.
4. Remove a Pasteur pipette from its container and attach a bulb to it.
5. Obtain a tube of hot sterile agar, and make two agar cushions per sterile slide using the sterile Pasteur pipette. If the agar starts to solidify, transfer the agar tube to a hot dry …show more content…
Place one slide under the microscope, noting down whether it is the experimental or control slide, locate the hyphal tip. Align the hyphal tip with the ocular micrometer in the direction of the polarized cell movement, this will allow the growth of the tip to be measured.
6. Measure and record the speed with which the hyphal tip is growing using the ocular micrometer to measure the distance and a stop watch to measure the time. You can arbitrarily pick the distance for which you wish to measure the cell movement. In this entire experiment, the distance of 10 ocular units measured in the increments of 2 ocular units will be used.
7. While the hyphal tip is still visible under the bright field phase, switch the microscope to Fs14 filter clock and turn the light intensity of the fluorescence to the maximum. You may have to shield your eyes from the lights in the classroom.
8. After the light intensity is adjusted accordingly, the ocular micrometer and fluorescent nuclei in red will be visible. The fluorescent nuclei are visible because N. crassa has an extra gene that makes the nucleus fluorescent.
9. Record where the nuclei are positioned in comparison to the tip during growth using the ocular
1. Start with wearing gloves and swabbing them with 70% ethanol to keep the materials that will be used in the experiment sterile.
2. Wipe the work station with Wescodyne and paper towels. It is important to keep the sterile materials such as slides in petri dish, Pasteur pipette in container and forceps covered as much as possible.
3. Place four to five filter papers in the petri dish and drip few drops of water on the strips, this will maintain moisture in the agar cushions.
4. Remove a Pasteur pipette from its container and attach a bulb to it.
5. Obtain a tube of hot sterile agar, and make two agar cushions per sterile slide using the sterile Pasteur pipette. If the agar starts to solidify, transfer the agar tube to a hot dry …show more content…
Place one slide under the microscope, noting down whether it is the experimental or control slide, locate the hyphal tip. Align the hyphal tip with the ocular micrometer in the direction of the polarized cell movement, this will allow the growth of the tip to be measured.
6. Measure and record the speed with which the hyphal tip is growing using the ocular micrometer to measure the distance and a stop watch to measure the time. You can arbitrarily pick the distance for which you wish to measure the cell movement. In this entire experiment, the distance of 10 ocular units measured in the increments of 2 ocular units will be used.
7. While the hyphal tip is still visible under the bright field phase, switch the microscope to Fs14 filter clock and turn the light intensity of the fluorescence to the maximum. You may have to shield your eyes from the lights in the classroom.
8. After the light intensity is adjusted accordingly, the ocular micrometer and fluorescent nuclei in red will be visible. The fluorescent nuclei are visible because N. crassa has an extra gene that makes the nucleus fluorescent.
9. Record where the nuclei are positioned in comparison to the tip during growth using the ocular