I am carrying out an experiment on Trichoderma and need to measure OD of the fungal growth after 7 days. Unlike bacteria, fungus clump together making It difficult to get OD for the fungal growth. Any optimized protocol or guidance on how can i can measure OD of the fungal growth in PDB. Thanks in advance.
Harvest Fungal Culture After 7 days of growth, harvest the fungal culture by filtering it through sterile cheesecloth or using a centrifuge to separate the fungal biomass from the broth. Resuspend the fungal biomass in a small volume of sterile saline or buffer. Add sterile glass beads to the resuspended fungal biomass. Vortex the mixture vigorously for 2-3 minutes to break up the clumps and create a uniform suspension. Immediately measure the OD of the homogenized suspension using a spectrophotometer at the appropriate wavelength (usually 600 nm for fungal biomass). Resuspend the fungal biomass in sterile saline or buffer. Use an ultrasonic homogenizer to sonicate the suspension for a few minutes with short pulses to avoid overheating. This will break up the clumps and create a uniform suspension. Measure the OD of the sonicated suspension immediately. Harvest the fungal culture. Resuspend the fungal biomass in sterile saline or buffer. Place the suspension on a magnetic stirrer with a stir bar. Stir continuously to keep the biomass evenly distributed. While stirring, take aliquots to measure the OD. This ensures a more consistent and representative reading. If measuring OD proves too challenging, consider measuring the dry weight of the fungal biomass as an alternative. Harvest the fungal culture, filter or centrifuge to collect the biomass, and dry it at 60–80 °C until a constant weight is achieved. Measure the dry weight using a precision balance. Ensure consistent sampling and handling to avoid variations in OD measurements.Calibrate your OD measurements by correlating them with dry weight or colony-forming units (CFUs) to get a more accurate assessment of fungal growth. Perform measurements in triplicates or more to account for variability.
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