Description and Discussion

The 96‑well microplate reference contains inorganic fluorescent materials in a sintered glass matrix and is available in both top and bottom reading configurations.

Plate dimensions comply with international standards for biomedical screening.

The inorganic fluorophores used are inherently stable and do not undergo photodecomposition “bleaching”, and solvent evaporation is not an issue because the references are solid-state.

Four different fluorophores, identified by the wavelength of their principal emission peak, provide a range of excitation and emission wavelengths to match the user’s application.

Typical spectra

TYPICAL SPECTRA

The “typical spectra” shown above were recorded on a representative filter‑based microplate reader with relatively broad excitation/emission bandwidths and are not spectrally corrected for the instrument’s wavelength‑dependent detector response.

They are intended to illustrate the approximate excitation and emission regions for each plate under typical plate‑reader conditions. Note that uncorrected spectra obtained on individual instruments will vary.

The Certificate of Validation supplied with each reference plate provides relative intensity values for all wells on the plate and the calculated coefficient of variation for those readings. These data support the routine qualification of fluorescence‑based plate readers. Typical data (for a bottom-reading BR615 plate) are shown below:

For users requiring a fully traceable, spectrally corrected emission spectrum certified and calibrated under ISO 17025. A Certified Reference Material (CRM) version of these plates is available, accompanied by a separate ISO/IEC 17025 Certificate of Calibration. Please see the Corrected Spectra Datasheet.

Corrected Spectra

ISO/IEC 17025 CERTIFIED REFERENCE MATERIAL (CRM) OPTION

Starna doped‑glass microplate references are also available as ISO/IEC 17025 accredited Certified Reference Materials, providing spectrally corrected, high‑resolution emission spectra for instrument calibration, spectral correction and method transfer. In this configuration, the plates are calibrated in the Starna ISO/IEC 17025 accredited calibration laboratory, with full traceability to primary standards.

For the ISO/IEC 17025 CRM plates, the emission spectra are measured on a reference‑grade spectrofluorometer with narrower excitation/emission bandwidths and corrected for the instrument’s wavelength‑dependent responsivity. The resulting certified spectra (shown below) represent the true relative emission distribution of the fluorescent materials, independent of any specific instrument, and provide more accurate band shapes and relative intensities, particularly in the red and near‑infrared regions where detector response typically declines.

By contrast, the “typical spectra” shown for the main product above are uncorrected spectra recorded on a filter‑based plate reader and are intended only as illustrative plate‑reader‑style response curves.

Each ISO/IEC 17025 CRM plate is supplied with a Certificate of Calibration and Traceability that states the certified, spectrally corrected emission spectrum over the working range, together with the associated measurement uncertainty and the conditions of validity. The certified values are traceable to a system of primary standards through an unbroken chain of calibrations in accordance with ISO/IEC 17025 and ISO 17034.

For a detailed explanation of why fluorescence emission spectra from plate readers are instrument‑specific, and how spectrally corrected Certified Reference Materials can be used to derive instrument response functions and correct routine measurements, see Technical Note CRM‑TN‑001: Certified Reference Materials for the Correction of Fluorescence Relative Emission Intensity Spectra.

Technical Note

This document includes worked examples using the TR‑800 doped‑glass microplate reference and illustrates the differences between uncorrected plate‑reader spectra and traceably corrected emission spectra.

CORRECTED SPECTRA

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