64206 2025 eng poster 0 -- -- -- The standardization and calibration of fluorescence microscopy have become increasingly vital due to the wide-spread use of in life and materials sciences. As the demand for reliable and user-friendly methods to assess micro-scope performance grows, universal calibration tools accessible to both researchers and vendors are needed. To support the standardization of characterization methods in microscopy, it is crucial to provide calibration tools together with standardized operating procedures for their effective implementation. The public-funded project "FluMiKal"* develops calibration tools in the shape of typical microscopic slides to assess key parameters such as spatial resolution, point spread function, spectral sensitivity, linearity and sensitivity of the detection system. The focus is on creating calibration tools that are user-friendly, robust, and versatile in their application. This work addresses the critical parameter of wavelength-dependent spectral sensitivity, which affects the meas-ured signals from the instrument side, yielding instrument-specific data and instrument aging-induced changes over time. For this purpose, μ-slides from ibidi with six channels are used, allowing them to be filled with different solutions containing molecular or nanoscale fluorophores with well-characterized absorption and fluorescence properties. The certified spectral fluorescence standards BAM-F003, F004, F005, and F007 assessed provided as ethanolic solutions by the Federal Institute for Materials Research and Testing (BAM), cover a broad spectral range from the blue to the near-infrared [1], [2]. Dye-based slide prototypes have been used to determine the spectral sensitivity of confocal microscopes from different vendors with various detector types by acquiring the spectral data of the BAM dyes under standardized measurement conditions, demonstrating the applicability of this concept. Proof-of-concept experiments could demonstrate the proper sealing of the slides. Further experiments will explore long-term stability and their potential as standards for relative intensity calibrations. * FluMiKal is funded by the Federal Ministry for Economic Affairs and Climate Action, Germany (WIPANO FKZ 03TN0047B) [1] doi: 10.1007/4243_2008_028. [2] doi: 10.1007/s00216-024-05723-w. European Light Microscopy Initiative - ELMI 2025 Heidelberg, Germany 03.06.2025 publish false true Ute Resch-Genger A. Fallisch E. P. Petrov R. Salhany C. Forthman Z. Guttenberg R. Nitschke eng uncontrolled Fluorescence eng uncontrolled Advanced material eng uncontrolled Validation eng uncontrolled Calibration eng uncontrolled Method comparison eng uncontrolled Reference material eng uncontrolled Dye eng uncontrolled Fluorescence standard eng uncontrolled Microscopy eng uncontrolled CLSM eng uncontrolled Imaging Analytische Chemie Ingenieurwissenschaften und zugeordnete Tätigkeiten 1 Analytische Chemie; Referenzmaterialien 1.2 Biophotonik Umwelt Chemie und Prozesstechnik Chemische Charakterisierung und Spurenanalytik Datei im Netzwerk der BAM verfügbar ("Closed Access") Präsentation