Why is serial dilution more accurate

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In a mix roughly in the middle of the well volume, the dispensed liquid is forced toward the well bottom while dispensing, and aspirated liquid is pulled from the center of the well. If the mix occurs close to the bottom of the plate, the dispensed liquid is pulled back into the tip during the aspiration. Mixing in the center allows the dispensed liquid to be more evenly distributed in the sample, thus increasing the likelihood of efficient mixing.

The VWorks software controlling the Bravo platform allows the creation of liquid classes, which allows the operator to modify the velocity and acceleration for aspirating, dispensing, and mixing tasks. Precision and accuracy improved as the mix velocity increased. The cause of this is likely due to the creation of more turbulent mixing, which in turn distributed the fluorescein dye more quickly throughout the solution.

Finally, the effect of dynamic tip retraction and extension was explored. This function moved the tips deeper into the well during each aspirate step, and retracted them during each dispensing step.

This allowed a larger volume of the well to be affected by the mix step by adding the movement of the tip into the mix task. There was a marginal improvement less than 0.

Additionally, no effect was observed by utilizing another mix standard, which involved aspirating close to the bottom of the well and dispensing near the top of the solution. This mixing method caused no improvement once the other parameters described above had been optimized.

These experiments mixed homogenous solutions; there may be an improvement with this technique if the solutions are expected to have different viscosities. Based on these experiments, the parameters that had the largest impact on efficient mixing were in decreasing order :. To verify this conclusion, the first experiment varying the number of mix cycles was repeated with the improved mix parameters.

The new parameters provided increased precision and accuracy and improved the accuracy and precision of the 3-mix cycle operation to a level comparable with the mix cycle operation Figure 2. More importantly, the new parameters also decreased the time required to run an effective serial dilution protocol from 20 minutes to just under 5 minutes.

This has tremendous potential in automating a serial dilution assay and ensuring accurate and precise results. Log in to leave a comment. Sign in. Forgot your password? Get help. Privacy Policy. Dilution can also be achieved by mixing a solution of higher concentration with an identical solution of lesser concentration. Diluting solutions is a necessary process in the laboratory, as stock solutions are often purchased and stored in very concentrated forms.

For the solutions to be usable in the lab for a titration, for instance , they must be accurately diluted to a known, lesser concentration. The volume of solvent needed to prepare the desired concentration of a new, diluted solution can be calculated mathematically. The relationship is as follows:. M 1 denotes the concentration of the original solution, and V1 denotes the volume of the original solution; M 2 represents the concentration of the diluted solution, and V2 represents the final volume of the diluted solution.

When calculating dilution factors, it is important that the units for both volume and concentration are the same for both sides of the equation.

Serial dilutions involve diluting a stock or standard solution multiple times in a row. Typically, the dilution factor remains constant for each dilution, resulting in an exponential decrease in concentration.