Measuring Microbial Growth

Lab 6
 Measuring Cell Growth
• Number of Cells per milliliter of liquid
– cells/ml
• Bacterial cultures contain millions of cells
• Direct Vs. indirect counts
– Direct Counts mean actually counting cells
• More accurate, but takes a lot longer
– Indirect Counts mean estimating the number of
cells based on turbidity in liquid or dry weight.
• Less accurate, but quick
• Total Count vs. Viable count
– Total countall cells in a culture
– Viable countonly live cells
 Turbidity Method
• Spectrophotometer
• Measure % transmittance
– Amount of light going through culture
– Transmittance is inversely proportional to
turbidity (cell concentration)
• As turbidity increases cell number increases and
transmittance decreases
• Indirect and Total Count
Turbidity and Transmittance
 Plate Counts
• Direct Viable counts
• The idea is to evenly spread the culture
across a plate and count the colonies
• Each colony represents a cell in the
original culture.
• Problem: If we have a culture that has
1x106 cells/ml and we put 1 ml on the
plate will we have isolated colonies?
 Plate Count
• We must dilute first
• If we put 1ml of culture into 9 ml of sterile media
we have performed a 1/10 serial dilution (SD)
• Formula: SD=AT/TV
– AT=amount of culture transferred
– TV=Total volume after transfer (1ml + 9 ml)
• What if we did 5 serial dilutions on that culture
that had 1 x 106 cells/ml