The facility takes care of three fluorescence confocal microscopes and provides user support. We offer standard confocal imaging and the acquisition of time series, tile scans, or z-scans. All biological experiments can be performed in a fully incubated regime, allowing an extended time of measurement. Apart from that, we also offer more advanced fluorescence microscopy techniques such as fluorescence lifetime imaging (FLIM), correlation spectroscopy (FCCS), anisotropy measurements, or the use of the Airyscan detector offering a better spatial resolution. Apart from confocal imaging, also cameras for wide-field experiments are available.

Our microscopes

1. ZEISS LSM 780 (laboratory C.3.08)

   The confocal microscope is equipped with the Intune laser with tunable wavelength (488 – 640 nm), 405 nm CW/pulsed laser, and with common CW lasers: Argon (458, 488, 514 nm), solid-state (561 nm), and HeNe laser (633 nm). It uses three internal PMT detectors, one of which is the GaAsp array detector for spectral imaging.

   Further instrumentation/techniques available:

   • FLIM with external SPAD detectors (Picoquant)
   • Correlation spectroscopy
   • Anisotropy measurements
   • Widefield imaging (TIRF included).

   
   
   
   

2. ZEISS LSM 980 (laboratory A.01.85)

   The confocal microscope is well suited for long-lasting biological experiments. It is therefore fully incubated (temperature, CO2). The microscope is equipped with standard laser lines (405, 445, 488, 561, 639 nm), spectral detection, and the Airyscan detector.
   Airyscan detector is an array detector that allows for analysis of the emission beam. It results in better spatial resolution (by factor 1.4), better signal-to-noise ratio, and/or faster acquisition. The fast acquisition mode is particularly beneficial when combined with a fast, cooled galvo scanner.
   
   

3. LEICA SP8 + FALCON upgrade (laboratory A.01.85)

   The confocal microscope is meant for more advanced (not only) biophysical experiments. It is therefore equipped with a battery of synchronizable pulsed lasers (405, 440, 470, 640 nm a white-light laser, covering a broad range of wavelength 470-670 nm) and with Leica Falcon FLIM system. It uses the so-called „fast FLIM“ approach that allows for significantly higher acquisition speed at the expense of temporal resolution (97 ns). Therefore, it enables the acquisition of FLIM at a video rate.
   Apart from FLIM, Falcon also enables fluorescence crosscorrelation spectroscopy measurements (FCCS), or its combination with pulsed interleaved excitation (PIE), which removes artifacts of the method based on nanosecond switching between two laser lines.

   
   
   

People

• Jana Humpolíčková (C.4.02, ☎ 477, jana.humpolickova@uochb.cas.cz)
• Petro Khoroshyy (A.01.77, ☎ 448, petro.khoroshyy@uochb.cas.cz)

Booking

Guidelines