The small size of bacteria cells makes an analysis in comparison to other cell types more difficult. Image-based methods fail in the measurement of bacterial cell concentrations and the simultaneous determination of cell counting and viability determination is rare. Amphasys offers here a technology which allows the determination of viability and cell count in parallel – without staining, markers, and incubation time.
The two main variables indicating the changes in bacteria are cell number and cell viability. In addition to providing these results, the high sensitivity of Impedance Flow Cytometry can be used to analyze a wide range of bacterial species, including both gram-negative and gram-positive bacteria. It can also be used to monitor bacterial growth, detect bacterial contamination, and measure bacterial viability.
· One instruments – a large range of cell sizes
· Simultaneous result of cell count and cell viability – among other cell properties
· label free, no staining procedures, no markers needed, no incubation
· short sample preparation and measurement times
· highly sophisticated software for data analysis
· Low operational cost
· No impact of turbidity and particles
· No calibrations, no adjustments
Case study: Inactivation of bacteria
To monitor the inactivation of bacteria, measurements with the Ampha X30 can easily be performed, measuring both, cell viability and cell count in parallel. The treatment of a E. coli culture with 70% ethanol on one hand and a culture heated for 30 min at 95°C on the other hand, shows the shift of the viable cells to the area of the dead cells. On the top left of each scatter plot the clouds of 1,5 mm PBS beads are detected which are much bigger than the bacteria cells.
The inactivation of bacteria can be monitored easily, also in a time resolved way. The scatter plots show that a certain part of the bacterial culture can be inactivated by the respective treatment and that results are easily available.
Case study: Efficacy of antibiotic compounds
Amphasys’ technology enables to determine the efficacy of antibiotic compounds on bacterial cultures quickly and easily. To carry out the measurement, neither markers nor staining dyes are needed, and an incubation overnight is not necessary neither.
An unknown strain of E. coli was prepared in three samples and spiked with 1.5 mm PBS beads (top left of each scatter plot). Ampicillin was added in two different quantities: 1.6 mg and 160 mg. The samples were measured 1 hour, 3 hours and 25 hours after the treatment. The scatter plots only display the viable cells.
The Control sample without Ampicillin shows high bacteria concentration throughout the duration of the trial period (top row). The small concentration of Ampicillin shows only a slight reduction on the bacterial cell concentration. The larger dose of Ampicillin is effective already after one hour and reduced the cell concentration effectively in the trial period.
The figure shows the cell concentration of the bacteria and the high efficacy of the higher dose of Ampicillin in comparison to the lower dose. Amphasys’ technology therefore helps to find the right treatment in an easy, very fast and accurate way.
Amphasys Cell Analyzer
The Ampha X30 is Amphasys’ next generation for cell analysis in Bioprocessing. It is tailor made for analyzing a broad range of different cell types like bacteria, mammalian, insect, algae or yeast. The newly designed technological platform enables highly accurate determination of, cell viability, concentration, differentiation, and cell status. This information must be considered in scale-up process during propagation in biofuel, bioplastic, enzyme, pharmaceutical or ethanol production.