INFO & USER GUIDE

Numerical INner Filter Effect Corrector - NINFE

by Tomislav Friganović, Davor Šakić and Tin Weitner

University of Zagreb, Faculty of Pharmacy and Biochemistry, Zagreb, Croatia.


Help from Mario Gabričević (discussion), Tino Šeba, Valentina Borko, and Robert Kerep (testing) is greatly appreciated. Please report all suggestion, changes, improvements, and usage to davor.sakic@pharma.unizg.hr.


Finanancial support provided through Croatian Science Foundation grant no. UIP-2017-05-9537 "Glycosylation as a factor in the iron transport mechanism of human serum transferrin" GlyMech at University of Zagreb Faculty of Pharmacy and Biochemistry.

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.

DISCLAIMER:
This website is in DEMO mode, in the hope that it will be useful,
but WITHOUT ANY WARRANTY; without even the implied warranty of
MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE.
Authors assume no responsibility or liability for any errors
or omissions in the content of this site. The information contained
in this site is provided on an "as is" basis with no guarantees of
completeness, accuracy, usefulness or timeliness.
USE this website, associated program, and results at
AT YOUR OWN RISK.
©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.