growth of bacteria Flashcards
what are four methods to estimate the growth of a bacterial culture
- Haemocytometer
- Turbidity
- Dilution plating
- Dry mass
Describe how to conduct haemocytometer ?
- Dilute broth sample with equal volume of trypan blue to stain dead cells
- Use a calibrated haemocytometer with volume 0.1mm3. Count the cells in each of the sets of 16 squares and calculate means
- Number of bacterial cells = number counted x 10 to the power of 4 per cm3
Is haemocytometer viable count or total count ?
total
does haemocytometer count the exact no. of cells ?
yes - human error
how fast is the haemocytometer ?
slow
what are the uses of haemocytometer ?
bacteria
explain how to conduct a turbidity ?
- Use colorimeter. Measure absorbable or % transmission of samples of known microorganism counts
- Plot calibration curve
- Record absorbable/ % transmission of unknown sample. The cloudier, the less light is absorbed. Plot graph
Is turbidity viable count or total count ?
total
does turbidity count the exact no. of cells ?
no
how fast is the turbidity ?
fast
what are the uses of turbidity ?
bacteria
explain how to conduct dilution plating
- Start with 10cm^3 of undiluted culture
- Add 1cm^3 of first tube into 9 cm^3 of sterile saline
- Continue to add 1cm^3 from previous tubes to form serial dilutions
why is appropriate dilution a compromise when diluting a culture?
Must have a low enough number of microorganisms to count and a high enough number to provide a reliable sample of the culture
is dilution plating viable or total count ?
viable
does dilution plating give the exact no. of cells ?
yes - human error