Introduction
This is a quick guide in using Numerical INner Filter Effect Corrector - NINFE.Although different methods are available for countering IFE, this method can be implemented into laboratory workflow with minimal hassle.
User guide
Necessary steps:
1) Determine the geometric parameter for the respective microplate reader and microplate type.2) Measure fluorescence for known fluorophore concentration series to be used for NINFE calibaration. At least two fluorescence measurements at two different z-axis positions (z-positions) for the same sample in the microplate well are required.
3) Format calibration data table (see details below).
4) Run NINFE calibration routine to obtain optimal z-position combination and linear response parameters.
5) Measure fluorescence for samples with unknown fluorophore concentration at the determined optimal z-position combination.
6) Format concentration readout data table (see details below).
7) Run NINFE readout routine to retrieve unknown fluorophore concentrations. The NINFE calibration routine needs to be performed previously. Readout can be used multiple times while using the same calibration.
8) Click START AGAIN for another cycle.
Use ACF in order to compare corrections using absorbance measurements in transparent 96-well plates.
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©Friganovic, Sakic, Weitner 2020-2021.
Data formats (input/output)
Input format (calibration):Output format (calibration):
Input format (readout):
*Designation = user-defined or sample-specific label, e.g. well position etc.
Output format (readout):
-Parametric model (Pm)
-Black-box model (BBm)
CREDITS
Idea: Tin Weitner, 2020.-2021.
Created by: Tin Weitner, Tomislav Friganović, and Davor Šakić, 2021.
Python version: Tomislav Friganović, 2021.
Web adaptation by: Davor Šakić, 2021.
Funding: HrZZ, GlyMech, 2021.