Overview of the measurements available on the SpICy technical platform.

The SAFAS plate reader is a commercial instrument that allows measurements of absorbance, fluorescence, or luminescence in both kinetic and spectral modes. This instrument is equipped with temperature regulation and sample agitation functions, and also offers the possibility to work in a spectroscopic cuvette.

SAFAS plate reader and an example of kinetic data.

LOLITA is a setup designed to acquire fluorescence emission decay data with high statistical and temporal quality in the 0.1-100 ns range, on solutions or powder/solid samples. The excitation source is a white laser (440-650 nm) and UV LEDs. Polarizers in the optical path allow the recording of fluorescence anisotropy decay, providing insights into the hydrodynamic volume of the emissive species or information about the oligomeric nature of a fluorescent protein.

LOLITA setup and fluorescence emission decay of yellow fluorescent proteins, with the IRF shown in gray.

BEAM is a setup that enables controlled irradiations (in terms of time and power), the triggering, and analysis of photoreactions. It facilitates simultaneous acquisition of absorption and fluorescence emission spectra (210-700 nm) and kinetic monitoring. Currently used to study the photoreactions of fluorescent proteins, it is open to other areas of research.

BEAM setup and examples of kinetic tracking of absorption and fluorescence emission spectra of a yellow fluorescent protein under irradiation.

FloRa is a wide-field microscope that allows dynamic multicolor or ratiometric imaging.

• • • • • • • • 2 objectives: x63, x40
              • Temperature regulation available at 37°C
              • LED light source: 380-680 nm
              • 6 available fluorescence spectral selections

FloRa microscope and an example of a study on new fluorescent probes for H2O2 detection in collaboration with ICSN and ICMMO.

BiFluor enables measurements of fluorescence intensity, fluorescence lifetime in imaging mode (FLIM), and fluorescence emission spectra under a microscope (for cell cultures, powders, solutions, or films) using the same instrument.

• • • • • • • • 2 objectives: x60, x20
              • Temperature regulation possible between 15-40°C
              • LED light source for intensity measurements: 380-680 nm
              • Pulsed diodes: 440 nm & 466 nm for lifetime measurements
              • PicoQuant detection, SymPhoTime64 software

BiFluor microscope with FLIM measurements on the left and fluorescence emission decay on films on the right.

Managed by Hadrien Jalaber, the SpICy platform is equipped with a Cytoflex, a multi-color cytometer from Beckman Coulter, featuring 3 excitation lasers (405, 488, 561 nm) and over 10 optical detection parameters. It can detect objects ranging from 1 to 100 µm. An analysis station with FloJoW software is available, and a FRET analysis strategy through cytometry (CytoFRET) is currently under development. This equipment is located near the cell culture room and various laboratory equipment.

Users can operate the cytometer with assistance from a platform engineer or independently after completing training. External users contribute to the maintenance and operational costs of the cytometer.

Beckman Coulter multi-color cytometer, schematic of its function, and example measurement.