Case Study
Rapid 96-well plate normalisation to a 10 µg/mL
Veon Scientific evaluated the i.prep 2 on a full 96-well plate normalisation workflow, bringing a variable-concentration source plate to a defined 10 µg/mL target using an independent 8-channel transfer sequence. By allowing each channel to execute its own calculated sample and diluent volume, i.prep 2 can automate true per-well normalisation in a streamlined plate-wide workflow, completing the process in around 5 minutes, compared with roughly 25 minutes on a conventional multi-channel system.
At a glance
Using runtime concentration data supplied as a worklist in Flow Studio, i.prep 2 normalised a full 96-well plate to a 10 µg/mL target in an independent 8-channel workflow. The process was completed in around 5 minutes, compared with roughly 25 minutes for a conventional multi-channel approach, while the final plate achieved a mean concentration of 10.98 µg/mL with a 2.48% CV. The result highlights a practical advantage of i.prep 2's independent channels: true per-well normalisation at plate scale, without sacrificing throughput.
Key takeaways
Results
Target concentration
10 µg/mL
Plate format
96 wells
Mean final concentration
10.98 µg/mL
Median final concentration
10.98 µg/mL
Standard deviation
0.27 µg/mL
CV
2.48%
Within +/-15% of target
96 / 96 wells
Within +/-20% of target
96 / 96 wells
The result is both tight and broad-based. Mean and median align closely, the total spread remains narrow at 0.27 µg/mL, and the full plate stays inside the stated tolerance bands. For a normalisation workflow, that matters because the system is not repeating one fixed dispense across the plate; it is coordinating well-specific volumes while still delivering a clean full-plate outcome.
The challenge
Normalising a plate sounds simple in principle, but in practice it quickly becomes a tedious and error-prone preparation step. When every well starts at a different concentration, each one may require a different sample and diluent volume to bring the full plate to a common target.
That is exactly where conventional multi-channel systems can become limiting. They are highly effective when every channel is repeating the same transfer pattern, but plate normalisation often requires well-specific volume adjustments across the plate. Instead of a simple repeated dispense, the workflow becomes a coordinated set of different transfers that still has to be executed accurately and consistently at throughput. i.prep 2 is designed to support that kind of application, helping automate plate normalisation across a full 96-well format without turning it back into a manual preparation task.
The workflow
Step 1
Load the runtime CSV in Flow Studio
Flow Studio is used to provide the protocol with the measured input concentrations for the plate at run time. That allows the same workflow to be reused with different plates while keeping the transfer plan tied to real sample data.
Step 2
Calculate per-well normalisation volumes
The protocol uses that runtime CSV to calculate the sample and diluent volume needed for each well to reach a 10 µg/mL target at a 120 uL final volume, creating a true per-well normalisation plan rather than one shared dispense recipe.
Step 3
Dispense diluent into the destination plate
Diluent is dispensed from Zone2 into the destination plate in Zone6 using an 8-channel batched workflow. This makes it practical to pre-load each destination well with the specific make-up volume it needs while still maintaining multi-channel throughput.
Step 4
Transfer sample into the prepared plate
Sample is then transferred from the source plate in Zone3 into the destination plate in Zone6 using the calculated per-well sample volumes. Fresh tips are picked up for each 8-well batch, allowing the workflow to stay efficient while handling different transfer volumes across the plate.
Plate heatmap of final concentration
Per-well final concentrations across the full 96-well plate
The heatmap shows a tightly clustered full-plate result, with concentrations remaining within the stated tolerance band across all 96 wells. Rather than drifting by row, column, or position, the plate converges into a consistent final range — exactly the outcome you want from a normalisation workflow where each well requires its own calculated transfer volume.
Ranked concentration profile against target band
Every well remains inside the shaded +/-15% operating band
Ranked across all 96 wells, the concentration profile remains compact from minimum to maximum. The minimum well measures 10.01 µg/mL, the maximum reaches 11.48 µg/mL, and the full plate stays inside the +/-15% target band even though the workflow is executing well-specific transfer volumes rather than one uniform dispense.
Why it matters
For assay teams and integrators, this kind of result matters because it shows a realistic full-plate normalisation workflow rather than a narrow spot check. It gives a clear indication that the platform can prepare plates to a controlled concentration target before downstream assay steps, while still accommodating well-specific volume requirements across the same run.
A system with independently addressable channels is especially useful here. Instead of breaking the task into serial single-well corrections or simplifying the protocol around shared volumes, the i.prep 2 can keep the workflow plate-based while each channel aspirates and dispenses the volume required for its assigned well. That is a meaningful practical advantage for normalisation workflows.
What the data shows
The central message is consistency. Mean and median are aligned, the spread stays narrow, and every well lands within the specified tolerance range. Taken together, that points to a stable full-plate result rather than a mixed plate with isolated success.
In practical terms, this suggests that i.prep 2 can deliver reliable plate normalisation performance across a complete 96-well workflow using independent 8-channel liquid handling. That makes it well suited to applications where controlled inputs matter, but where each well still needs its own calculated transfer volumes to reach the same target.
If you want to discuss plate normalisation, assay setup, or how i.prep 2 fits into your liquid handling workflow, we can share more detail on the platform and its measured performance.