Institute of Molecular Genetics of the Czech Academy of Sciences
Inverted microscope DMI6000 with confocal head Leica TCS SP5 AOBS Tandem is a confocal laser scanning microscope in which all filtering and beam-splitting functions are performed by acusto-optical components, which makes the system extremely flexible. The tandem scanner allows to switch between standard or very fast resonant scanner, which is useful especially for dynamic processes in live cells. For that reason, the microscope is enclosed in climate chamber for work at 37°C and regulated CO2 atmosphere. The system is equipped with both FRAP and FRET modules.
| Methods | Confocal scanning fluorescence imaging Transmitted light imaging Brightfield and Nomarski contrast (DIC) (automated) Tile-scans & merge in LAS AF software Time-lapse experiments Multi-positions experiment Photo-kinetic experiment FRET (SE or Acceptor photobleaching) |
| Illumination | 405 nm diode laser, 50 mW 458, 476, 488, 496, 514 nm argon ion laser, 100 mW 561 nm diode-pumped solid-state DPSS laser, 10 mW 633 nm He/Ne red laser, 10 mW HLX 100 W halogen lamp for transmitted light HXP 120W/45C VIS Hg lamp – Leica EL6000 for fluorescence |
| Objectives | HCX PL APO 10x/0.40 DRY CS; FWD 2.2; CG 0.17 | BF, POL HC PL APO 20x/0.7 IMM CORR λBL; W/GLYC/OIL ; FWD 0.26; CG 0-0.17 | BF, POL, DIC HC PL APO 40x/1.30 OIL CS2; FWD 0.24; CG 0.17 | BF, POL, DIC HCX PL APO 63x/1.3 GLYC CORR 37°C; FWD 0.28; CG 0.14-0.18 | BF, POL HC PL APO 63x/1.40 OIL CS2; FWD 0.14; CG 0.17 | BF, POL, DIC |
| Eyepiece filtercubes | A (Ex: 360/40; DM 400; Em: LP 425) I3 (Ex: 470/40; DM 510; Em: LP 515) N2.1 (Ex: 537/45; DM 580; Em: LP 590) Y5 (Ex: 620/60; DM 660; Em: 700/75) |
| Detectors | 2x photomultiplier tube (PMT) 3x supersensitive hybrid detectors (HyD) 1x transmitted light detector |
| Confocal head | acousto-optical tunable filter (AOTF) acousto-optical beam splitter (AOBS) standard scanner (10-1400 Hz line frequency) resonant scanner (8000 Hz line frequency) maximum scanner resolution 8192×8192 pixels hardware zoom 1x-64x |
| Stage | motorized microscope stage with Super Z-galvo scanning insert for fast and precise Z movement |
| Aditional equipment | incubation unit (CO2, temperature) antivibration table |
| Software | LAS AF with FRET and FRAP modules |
| Location | room no. 0.172 (Green) |
| Phone | ext. 3172 |
| Booking | Calpendo (“SP5”) |
Inverted microscope DMi8 with confocal head Leica TCS SP8 is a confocal laser scanning microscope with full transmitted light equipment including transmitted light PMT and differential interference contrast (DIC) available for all objectives. Leica TCS SP8 it is a mirror-based system, using iterference filters for light filtering and beam-splitting. The laser power is modulated by acusto-optical tunable filter (AOTF).
| Methods | Confocal scanning fluorescence imaging Transmitted light imaging Brightfield and Nomarski contrast (DIC) (manual) Tile-scans & merge in LAS X software Time-lapse experiments Multi-positions experiment Photo-kinetic experiment FRET (SE or Acceptor photobleaching) |
| Illumination | 405 nm diode laser, 50mW 488 nm solid-state laser, 20 mW 552 nm solid-state laser, 20 mW 638 nm solid-state laser, 30 mW HLX 100 W halogen lamp for transmitted light HXP 120W/45C VIS Hg lamp – Leica EL6000 with for fluorescence |
| Objectives | HC PL FLUOTAR 10x/0.30 DRY; FWD 11.0 | BF, POL, DIC HC PL FLUOTAR 25x/0.75 OIL; FWD 0.15 | BF, POL HC PL APO 20x/0.75 IMM CORR CS2; FWD 0.66 | BF, POL HC PL APO 40x/1.30 OIL CS2; FWD 0.24; CG 0.17 | BF, POL, DIC HC PL APO 63x/1.40 OIL CS2; FWD 0.14; CG 0.17 | BF, POL, DIC |
| Eyepiece filtercubes | DAPI (A) (Ex: 360/40, Em: LP 425) FITC (I3) (Ex: 470/40; DM 510; Em: LP 515) RHOD (N2.1) (Ex: 537/45; DM 580; Em: LP 590) Cy5 (Y5) (Ex: 620/60; DM 660; Em: 700/75) CFP (Ex: 436/20; DM 455; Em: 480/40) |
| Detectors | 3x photomultiplier tube (PMT) 2x supersensitive hybrid detectors (HyD) 1x transmitted light detector |
| Confocal head | acousto-optical tunable filter (AOTF) low incident angle dichroic beam splitters standard scanner (1-1800 Hz line frequency) maximum scanner resolution 8192×8192 pixels hardware zoom 0.75x-48x |
| Dichroic mirrors | 488/552/638 nm tripple excitation dichroic 488/552 nm dual excitation dichroic Substrate RT 15/85 |
| Stage | motorized microscope stage with Super Z-galvo scanning insert for fast and precise Z movement |
| Aditional equipment | antivibration table |
| Software | LAS X |
| Location | room no. 0.172 (Green) |
| Phone | ext. 3172 |
| Booking | Calpendo (“SP8” ) |
Fully motorized inverted microscope DMi8 with confocal head Leica STELLARIS 8 equipped with wide-range while light laser with pulse picker (WLL PP) and highly sensitive time-resolved detection allows a variety of precise acquisition from standard confocal through very fast live cell acquisition up to large object imaging. The system is equipped with several acquisition modules for proceeding photo-kinetic experiments, FLIM or FLIM-FRET based experiments to the high resolution imaging followed with unique adaptive deconvolution module. The combination of AOBS and spectral sliders on detector cascade allows the fine tuning in selection of an excitation laser line and precise definition of emission spectra. For long-term live cell experiments are secured with hardware autofocus system (AFC). The system is equipped with incubation secures e.g. 37 °C and CO2 controlled chamber.
| Methods | Confocal scanning fluorescence imaging with Lightning superresolution module Transmitted light imaging Brightfield and Nomarski contrast (DIC) (automated) Tile-scans & merge in LAS X software Time-lapse experiments Multi-positions experiments Photo-kinetic experiments FRET (SE or Acceptor photobleaching) FLIM, FRET-FLIM experiments |
| Illumination | 405 nm cw diode laser, AOTF modulated, 100mW 440 – 790 nm Pulsed white light laser with pulse picker (WLL PP):
12V 100W LED source for transmitted light |
| Objectives | HC PL FLUOTAR 10x/0.30 DRY CS2; FWD 11.0 mm | BF, POL, DIC HC PL APO 20x/0.75 IMM CORR CS2; FWD 0.67 mm | BF, POL HC PL APO 86x/1.20 W CS2; FWD 0.30 mm| BF, POL, DIC HC PL APO 63x/1.40 OIL CS2; FWD 0.14 MM| BF, POL, DIC |
| Eyepiece filtercubes | DAPI (Ex: 355/56, DC: 400, Em: 460/50) GFP (Ex: 470/40; DC 495; Em: 525/50) RHOD (Ex: 546/10; DC 560; Em: 585/40) CFP/YFP (Ex: 435/25 + 500/20; DC 450 + 515; Em: 470/25 + 535/30) |
| Detectors | 2x supersensitive silicon-based hybrid detectors (HyD-S) 2x supersensitive hybrid detectors (HyD-X) for very fast time-resolve acquisition (FLIM) 1x supersensitive hybrid detector (HyD-R) for long wavelengths (700 – 850 nm) 1x transmitted light TL – PMT detector |
| Confocal head |
8-channels acousto-optical tunable filter (AOTF) for fast laser intensity modulation Conventional linear scanner (1-2600 Hz line frequency)
Resonant scanner (8000 Hz line frequency)
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| Time-resolve modules |
Tau-sense modules for time-resolved acquisition (time-resolved imaging without further analysis): TauGating, TauContrast, TauSeparation STELLARIS FALCON: FAST Lifetime CONtrast (FALCON) is solution for fast image acquisition, processing and FLIM analysis with modules:
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| Computational super-resolution module | Lightning – a super-resolution module based on mutual acquisition control and adaptive deconvolution based on SNR decision mask estimation. Maximal declared resolution is 120 nm laterally and 200 nm axially. |
| Stage | motorized microscope stage with Super Z-galvo scanning insert for fast and precise Z movement |
| Aditional equipment | antivibration table OKO-LAB incubation unit (CO2 + O2, temperature and humidity control system) |
| Software | All features are fully implemented to the LAS X software Las X modules:
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| Location | room no. 0.172 (Green) |
| Phone | ext. 3171 |
| Booking | Calpendo (“Stellaris”) |
Dragonfly is a fully motorized multi-modal imaging platform with confocal and widefield modes. The heart of the microscope is the confocal spinning disk which in combination with high-speed sCMOS and high-sensitive EMCCD cameras provides the system with exceptional speed and sensitivity. The low phototoxicity and photobleaching is ideal for live or delicate specimens. Dragonfly is 10-20 times faster than a traditional laser scanning confocal, which makes it optimal for imaging of fast dynamic events in live cell or for high throughput experiments. Photomanipulation experiments can be performed with Mosaic, a tool that allows for simultaneous illumination of multiple regions of interest of flexible shape and size.
The semi-automated microinjector suited for adherent cells microinjection is also available.
| Methods | Confocal spinning disk fluorescence imaging Transmitted light imaging Brightfield and Nomarski contrast (DIC) Large tile-scans & merge in Fusion software Time-lapse experiments with HW autofocus Multi-positions experiment Photo-kinetic and opto-genetic experiment (limited software solution) |
| Illumination | 405 nm solid-state laser, 200 mW 445 nm solid-state laser, 75 mW 488 nm solid-state laser, 150 mW 514 nm solid-state laser, 40 mW 561 nm solid-state laser, 100 mW 637 nm solid-state laser, 140 mW White LED source Leica DM for transmitted light CoolLED pE-300; 365 UV – white LED source for fluorescence 445 nm solid-state photo-manipulation laser for Mosaic, 1.3 W White LED source for Mosaic photo-manipulation (CoolLED pE-300; 365 UV) |
| Objectives | (HC PL APO 10x/0.40 DRY CS2; FWD 2.2; CG 0.17 | BF, POL, DIC) uppon request HC PL APO 20x/0.75 IMM CORR CS2; FWD 0.66 | BF, POL, DIC HC PL APO 40x/1.10 W CORR CS2; FWD 0.65; CG 0.14-0.18 | BF, POL, DIC HC PL APO 63x/ 1.20 W CORR CS2; FWD 0.3; CG 0.14-0.18 | BF, POL, DIC HCX PL APO 40x/1.25-0.75 OIL λBL; FWD 0.13; CG 0.17 | BF, POL HCX PL APO 63x/1.40-0.6 OIL λB; FWD 0.12; CG 0.17 | BF, POL, DIC HCX PL APO 100x/1.4-0.7 Oil CS; FWD 0.13; CG 0.17 | BF, POL, DIC |
| Dichroic mirrors | 405/488/561/640 nm Quad excitation dichroic 399-452/514/640 nm Triple excitation dichroic 405/488/561/685 nm Quad excitation dichroic |
| Eyepiece filtercubes | DAPI (Ex: 350/50; DC: 400; Em: 460/50) CFP/YFP (Ex: 435/25, 500/20; DC: 450; Em: 470/25, 535/20) FITC (Ex: 480/40; DC: 505; 527/30) RHOD (Ex: 546/10; DC: 560; 585/40) |
| Cameras | Zyla 4.2 PLUS sCMOS camera – 2048 x 2048 pix; 6,5 µm pixel iXon Ultra 888 EMCCD camera – 1024 x 1024 pix; 13 µm pixel |
| Dual camera switching mirror | LP 500 nm dichroic mirror (reflection to Zyla camera) LP 565 nm dichroic mirror (reflection to Zyla camera) 100% mirror (Zyla camera) 100% transmission dummy glass (iXon camera) |
| Zyla camera emission filter wheel | 450/50 nm bandpass filter (DAPI) 480/40 nm bandpass filter (CFP) 525/50 nm bandpass filter (GFP, A488, FITC) 540/30 nm bandpass filter (YFP) 600/50 nm bandpass filter (RFP, TRITC) 700/75 nm bandpass filter (Cy5) 405 to 445-515-561-730 quad emission filter (DAPI/CFP/YFP/mCherry/DyLight) Polarization filter |
| iXon camera emission filter wheel | 450/50 nm bandpass filter (DAPI) 480/40 nm bandpass filter (CFP) 525/50 nm bandpass filter (GFP, A488, FITC) 540/30 nm bandpass filter (YFP) 600/50 nm bandpass filter (RFP, TRITC) 620/60 nm bandpass filter (mCherry) 700/75 nm bandpass filter (Cy5) 405-488-561-640 quad emission filter (DAPI/GFP/RFP/Cy5) |
| Spare camera emission filter wheel | Polarization filter 725/40 nm bandpass filter 405-488-561-640 quad emission filter (DAPI/GFP/RFP/Cy5) 405 to 445-515-561-730 quad emission filter (DAPI/CFP/YFP/mCherry/DyLight) 445-515-561 tripple emission filter (CFP/YFP/RFP) 488-561 dual emission filter (GFP/RFP) 445-515 dual emission filter (CFP/YFP) |
| Microinjection | Microinjector FemtoJet joined with micromanipulator InjectMan NI 2 (Eppendorf) For volumes up to 100 pl Compressor to deliver the required pressure Integrated coarse and fine manipulator Work range: 20 mm per axis Automated and programmable axial injection movement |
| Aditional equipment | Okolab incubation unit including climate chamber (CO2, temperature, humidity) antivibration table photomanipulation module Mosaic (for FRAP, bleaching, uncaging, opto-genetics…) |
| Software | Fusion – acquisition software Imaris – visualization software Andor iQ3 – controlling Mosaic module |
| Location | room no. 0.171 (Orange) |
| Phone | ext. 3171 |
| Booking | Calpendo (“Dragonfly”) |
Upright routine widefield fluorescence microscope with motorized stage, fully automated transmitted and fluorescence light axis, and both monochromatic and color cameras. The monochromatic one is a high resolution very sensitive grayscale sCMOS camera. The LAS X Navigator tool allows to create high-resolution image tile-scans of large areas using the precise motorized stage.
| Methods | Routine fluorescence imaging True color RGB imaging in transmitted light mode Brightfield/Darkfield and Phase contrast Large tile-scans & auto merge in LAS X Navigator |
| Illumination | HLX 100 W Halogen lamp for transmitted light HXP 120W/45C VIS Hg lamp – Leica EL6000 for fluorescence |
| Objectives | HC PL FLUOTAR 5x/0.15 DRY; FWD 12 | BF, DF, POL HC PL APO 10x/0.40 DRY PH1; FWD 2.2; CG 0.17 | BF, DF, POL, PH HC PLAN APO 20x/0.70 DRY PH2; FWD 0.59; CG 0.17 | BF, DF, POL, PH HCX PL APO 40x/0.75 DRY PH2; CG 0.17 | BF, DF, POL, PH HCX PL APO 63x/1.40 OIL PH3 CS; FWD 0.1; CG 0.17 | BF, POL, PH HCX PL APO 100x/1.40-0.70 OIL; FWD 0.09; CG 0.17 | BF, DF, POL |
| Filtercubes | DAPI (A4) (Ex: 360/40; DM 400; Em: 470/40) FITC (L5) (Ex: 480/40; DM 505; Em: 527/30) TRITC (N3) (Ex: 546/12; DM 565; Em: 600/40) Cy5 (Y5)(Ex: 620/60; DM 660; Em: 700/75) |
| Cameras | Leica DFC490 – color CCD camera; 2,7 µm pixel Leica DFC 9000 – monochromatic sCMOS camera; 6,5 µm pixel, QE: min. 82 % |
| Stage | motorized microscope stage xyz |
| Software | LAS X, 64 Bit |
| Location | room no. 0.172 (Green) |
| Phone | ext. 3172 |
| Booking | Calpendo (“DM6000” ) |
Upright widefield fluorescence microscope with motorized stage, fully automated transmitted and fluorescence light axis and high-resolution very sensitive monochromatic 16-bit sCMOS camera. The LAS X Navigator tool allows to create high-resolution image tile-scans of large areas using the precise motorized stage.
| Methods | Routine fluorescence imaging Brightfield/Darkfield and Phase contrast Large tile-scans & auto merge in LAS X Navigator |
| Illumination | HLX 100 W Halogen lamp for transmitted light HXP 120W/45C VIS Hg lamp – Leica EL6000 for fluorescence |
| Objectives | HC PL FLUOTAR 6.3x/0.13 DRY; FWD 12.860 | BF, POL HC PL FLUOTAR 10x/0.30 DRY PH1; FWD 11.0; CG 0.17 | BF, DF, POL, PH HC PL FLUOTAR 20x/0.40 DRY PH1; FWD 7.5; CG 0.17 | BF, DF, POL, PH HC PLAN APO 20x/0.70 DRY PH2; FWD 0.59; CG 0.17 | BF, DF, POL, PH HCX PL APO 40x/0.75 DRY PH2; FWD 0.28 CG 0.17 | BF, DF, POL, PH HCX PL APO 63x/1.40 OIL; FWD 0.14; CG 0.17 | BF, POL, PH HCX PL FLUOTAR 100x/1.3 OIL PH3; FWD 0.13; CG 0.17 | BF, POL |
| Filtercubes | DAPI (A) (Ex: 360/40; DM 400; Em: LP 425) GFP (Ex: 470/40; DM 495; Em: 525/50) TRITC (N3) (Ex: 546/12; DM 565; Em: 600/40) Cy5 (Y5)(Ex: 620/60; DM 660; Em: 700/75) |
| Cameras | Leica DFC 9000 – monochromatic sCMOS camera; 6,5 µm pixel, QE: min. 82 % |
| Stage | motorized microscope stage xyz |
| Software | LAS X, 64 Bit |
| Location | room no. 0.172 (Green) |
| Phone | ext. 3172 |
| Booking | Calpendo (“DM6000-2”) |
Leica DM8 – Inverted widefield fluorescence microscope with motorized stage and fully automated transmitted and fluorescence light axis. The system is dedicated for live cell imaging. Long-term time-lapse live cell experiments are secured with hardware autofocus system. The microcope is also equipped with infinity port laser-scanner for precise photo-kinetic experiment (FRAP, iFRAP, Photo-activation…). All the features are fully implemented in LAS X software.
For long-term live cell experiments are secured with hardware autofocus system.
| Methods | Routine fluorescence imaging True color RGB imaging in transmitted light mode Brightfield/Darkfield, Phase contrast, Modulation contrast Large tile-scans & auto merge in LAS X Navigator Time-lapse experiments with HW autofocus Multi-positions experiment Photo-kinetic experiments |
| Illumination | LED light source for transmitted light lasers for photomanipulation:
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| Objectives | HC PL FLUOTAR 5x/0.25, FWD 12 | BF, POL, PH, IMC N PLAN 10x/0.25 DRY; FWD 17.6 | BF, POL, PH, IMC HCX PL FLUOTAR L 20x/0.40 DRY CORR; FWD 6.9; CG 0-2 | BF, POL, PH, IMC HCX PL FLUOTAR 40x/0.95 DRY CORR; FWD 3.3; CG 0-2 | BF, POL, PH, IMC HCX PL APO 40x/1.25 oil | BF, POL HCX PL APO 63x/1.40-0.60 OIL; FWD 0.1; CG 0.17 | BF, POLBF, POL |
| Filtercubes | DAPI (A4) (Ex: 350/50; DM 400; Em: 460/50) FITC (Ex: 480/40; DM 505; Em: 527/30) TRITC (Ex: 546/10; DM 560; Em: 585/40) Y5 (Cy5) (Ex: 620/60; DM 660; Em: 700/75) |
| Cameras | Leica DFC490 – color CCD camera; 2,7 µm pixel Leica DFC 9000 – monochromatic sCMOS camera; 6,5 µm pixel, QE: min. 82 % |
| Stage | motorized microscope stage xyz |
| Aditional equipment | PEACON incubation unit (CO2, temperature, humidity) |
| Software | LAS X |
| Location | room no. 0.171 (Orange) |
| Phone | ext. 3171 |
| Booking | Calpendo (“DMI 8” ) |
Inverted widefield fluorescence microscope DMI6000 with fully automated transmitted and fluorescence light axis. The transmitted light equipment includes differential interference contrast (DIC) prism. An incubation chamber makes the system suitable for live-cell imaging. The total internal reflection fluorescence illumination module (Leica AM TIRF MC) controls the angle of the excitation beam and makes use of the total reflection to excite only molecules in the thin section in contact with the coverglass. The system is equipped with both FRAP and FRET modules.
| Methods | Routine fluorescence imaging Brightfield, Nomarski contrast (DIC) Large tile-scans & auto merge in LAS X Navigator Multi-positions experiment Time-lapse experiments TIRF microscopy experiments |
| Illumination | HLX 100 W Halogen lamp for transmitted light HXP 120W/45C VIS Hg lamp – Leica EL6000 with for fluorescence solid-state TIRF laser 405 nm solid-state TIRF laser 488 nm solid-state TIRF laser 561 nm solid-state TIRF laser 633 nm |
| Objectives | HCX PL FLUOTAR 10x/0.30 DRY PH1; FWD 11.0 | BF, POL HCX PL FLUOTAR L 20x/0.40 CORR DRY PH1; FWD 6.9; CG 0-2 | BF, POL N PLAN L 40x/0.55 CORR DRY; FWD 3.3; CG 0-2 | BF, POL HCX PL APO 63x/1.3 GLYC CORR 37°C; FWD 0.28; CG 0.14-0.18 | BF, POL, DIC HCX PL APO 63x/1.40-0,60 OIL CS; FWD 0.1; CG 0.17 | BF, POL, DIC HCX PL APO 100x/1.46 OIL CORR CS; FWD 0.09; CG 0.1-0.22 | BF, POL, DIC |
| Filtercubes | A (Ex: 360/40; DM 400; Em: LP425) CFP-T (Ex: 422/44; DM 455; Em: 480/40) GFP-T (Ex: 490/20; DM500; Em: 525/50) YFP-T (Ex: 490/20, Em: 535/30) Cy3-T (Ex: 560/10, Em: 610/65) Cy5-T (Ex: 635/10, Em: 720/60) TRI (B/G/R) (ExFW: 420/30; 495/15; 570/20, Em: 457/22;530/20;628/28) FRET kostka pro CFP/YFP (ExFW: 436/12, 500/20; EmFW: 467/37, 545/45) QUAD-T (ExFW: 405/12, 490/20, 560/30,635/20; EmFW: 450/50, 525/36,600/32,705/72) |
| Cameras | Leica DFC350FX R2 – monochromatic CCD camera; 6,4 µm pixel Leica DF300 FX – color CCD camera; 6,4 µm pixel Andor iXon 897 – high sensitivity EMCCD back-iluminated camera, 512×512 px, suitable for single molecule live cell imaging; 16 µm pixel |
| Stage | motorized microscope stage with Super Z-galvo scanning insert for fast and precise Z movement |
| Aditional equipment | incubation unit (CO2, temperature, humidity) acousto-optical tunable filter (AOTF) – laser control |
| Software | LAS X – 64 bit; with FRET and FRAP and deconvolution modules |
| Location | room no. 0.171 (Orange) |
| Phone | ext. 3171 |
| Booking | Calpendo (“TIRF”) |
The system is built on the inverted widefield Olympus microscope version IX-71. The system is equipped with the HW autofocus system and an incubation chamber, and is suitable for multi-location and time-lapse experiments. The lasers for photomanipulation enable performance of photokinetics experiments (FRAP) and FRET. The sensitive camera makes the microscope suitable for live cell imaging and low fluorescence intensity applications.
| Methods | Routine fluorescence imaging Brightfield, Nomarski contrast (DIC) Tile-scans Multi-positions experiment Time-lapse experiments with HW autofocus Photo-kinetic experiments |
| Illumination | Lumencore LED – solid-state light source for fluorescence lamp for transmitted light lasers for photomanipulation (adjustable spot size):
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| Objectives | U PLAN FL 20x/0.50 DRY PH1; FWD 1.6; CG 0.17 | BF, POL U PLAN FL 40x/0.75 DRY; FWD 0.51; CG 0.17 | BF, POL U APO/340 40x/1.35-0.65 CORR OIL; FWD 0.1; CG 0.17 | BF, POL PLAN APO N 60x/1.42 OIL; FWD 0.15; CG 0.17 | BF, POL, DIC |
| Lumencore excitation filters | DAPI 390/18 CFP 438/24 GFP/FITC 475/28 YFP 513/17 RD-TR-PE 542/27 mCherry/Alexa594 575/25 Cy5 632/22 |
| Dichroic mirrors | Standard (DAPI, FITC, RD-TR-PE, Cy5) Live Cell (CFP, YFP, mCherry) Alexa (GFP, mCherry, Alexa 594) |
| Emission filter wheel | DAPI (435/48) FITC (523/36) RD-TR-PE (594/45) Alexa 594 (632/60) Cy5 (676/34) GFP (525/50) mCherry (632/60) CFP (470/24) YFP (559/38) |
| Eyepiece filers | the same as on emission filter wheel (excluding Cy5 filter) |
| Cameras | Photometrics CoolSANP HQ – high sensitive CCD camera, 1024×1024 px; 6,45 µm pixel |
| Stage | motorized stage with xyz movement equipped with HW AF |
| Aditional equipment | incubation unit (CO2, temperature, humidity) antivibration table hardware Ultimate autofocus for maintenance in-focus stability within the entire specimen |
| Software | SoftWorx
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| Location | room no. 0.171 (Orange) |
| Phone | ext. 3171 |
| Booking | Calpendo (“DVcore”) |
Scan^R is a module built on the inverted widefield microscope Olympus IX81. The system is designed for fully automated image acquisition and data analysis of biological samples. Scan^R can handle many different specimen formats, including multi-well plates, slides and custom-built arrays. The hardware autofocus maintains in-focus stability within the entire specimen. The acquisition is followed by complex image analysis and advanced data evaluation. Due to an incubation unit and a sensitive camera, the microscope is suitable for live cell imaging and low fluorescence intensity applications.
| Methods | High-content imaging (preferentially in widefield fluorescence mode) |
| Illumination | Xenon Mercury mixed gas arc lamp MT20 150 W (intense peaks at 365, 405, 436, 546, 577) lamp for transmitted light |
| Objectives | LUCPLFN 20x/0.45 DRY CORR PH1; FWD 6.6-7.8; CG 0-2 LUCPLFN 40x/0.6 DRY CORR PH2; FWD 2.7-4; CG 0-2 UPLSAPO 10x/0.4 DRY; FWD 3.1; CG 0.17 UPLFLN 40x/1.3 OIL; FWD 0.2; CG 0.17 UPLSAPO 40x/0.95 DRY CORR; FWD 0.18; CG 0.11 – 0.2 UPLSAPO 60x/1.35 OIL; FWD 0.15; 0.17 Optional Tube lens 1.6x |
| Excitation filters | DAPI 360/40 nm FITC 487/15 nm TRITC 552/25 nm GFP 470/20 nm HQ Cy5 623/55 nm CFP 435/10 nm YFP 500/20 nm |
| Emission filtercubes | DAPI/FITC/TRITC (460/10; 522/15; 605/30 nm) GFP HQ (518/45 nm HQ) Cy5 (705/35 nm) CFP/YFP (470/20; 545/50 nm) |
| Cameras | sCMOS camera Hammamatsu ORCA-Flash4.0 V2 – 6,5 µm pixel |
| Stage | motorized microscope stage xyz |
| Aditional equipment | incubation unit (CO2, temperature, humidity) antivibration table hardware autofocus |
| Software | ScanR acquisition ScanR analysis |
| Location | room no. 0.175 (Blue) |
| Phone | ext. 2750 |
| Booking | Calpendo (“ScanR”) |
Scan^R is a module built on the inverted widefield microscope Olympus IX81. The system is designed for fully automated image acquisition and data analysis of biological samples. Scan^R can handle many different specimen formats, including multi-well plates, slides and custom-built arrays. The hardware autofocus maintains in-focus stability within the entire specimen. The acquisition is followed by complex image analysis and advanced data evaluation. Due to an incubation unit and a sensitive camera, the microscope is suitable for live cell imaging and low fluorescence intensity applications.
| Methods | High-content imaging (preferentially in widefield fluorescence mode) |
| Illumination | Lumencore LED – solid-state light source for fluorescence (peaks: 395, 438, 475, 511, 555, 575 a 635 nm) Lamp for transmitted light |
| Objectives | UPLSAPO 10x/0.4 DRY; FWD 3.1; CG 0.17 UPLXAPO 20x/0.8 DRY CORR; FWD 0.6; CG 0 – 2 UPLXAPO 40x/0.95 DRY CORR; FWD 0.18; CG 0.11 – 0.2 UPLXAPO 60x/1.42 OIL; FWD 0.15; 0.15 Plan apochromatic 20x, NA 0.8, WD 0,6 mm |
| Excitation LED source | DAPI 395/25 nm, CFP 438/29 nm, FITC 475/28 nm, YFP 511/16 nm, TRITC 555/28 nm mCherry 575/25 nm Cy5 635/22 nm |
| Emission filtercubes | QUAD dichroic: DAPI/FITC/Cy3/Cy5 |
| Cameras | sCMOS camera Hammamatsu ORCA-Flash4.0 LT+90, pixel 6,5 µm pixel |
| Stage | motorized microscope stage xyz |
| Aditional equipment | hardware autofocus |
| Software | ScanR acquisition ScanR analysis |
| Location | room no. 0.175 (Blue) |
| Phone | ext. 2750 |
| Booking | Calpendo (“ScanR-2”) |
Zeiss Axiom Zoom.V16 – Fully motorized fluorescence macroscope with high resolution objectives is equipped with two cameras and a motorized 16:1 zoom with apochromatic correction. The system is well suited for a wide range of bigger samples and has superior brightness in large object fields. ApoTome.2 uses structured illumination to improve contrast and to increase resolution of the optical sections.
| Methods | Fluorescence imaging on widefield OR Apotome (optical sectioning) mode Transmitted and/or reflected light True color RGB imaging in transmitted light mode Brightfield/Darkfield, Opaque contrast Large tile-scans & auto merge in ZEN Blue software Time-lapse experiments Multi-positions experiment |
| Illumination | HXP 120 V – metal halid lamp for fluorescence CL 9000 LED CAN – transmitted light source CL 9000 LED CAN – incident light source |
| Objectives | PlanNeoFluar Z 1.0x/0.25; FWD 56 | BF, DF, oblique illumination + Reflected light PlanNeoFluar Z 2.3x/0.57; FWD 10.6 | BF, DF, oblique illumination + Reflected light |
| Fluorescence filtercubes | DAPI Filter set 49 (Ex: G 365; BS FT 395; Em: BP 445/50) eGFP Filter set 38 HE (Ex: BP 470/40; BS FT 495; Em: BP 525/50) Cy3 Filter set 63 HE (Ex: BP 572/25; BS FT 590; Em: BP 629/62) Cy5 Filter set 50 (Ex: BP 640/30; BS FT 660; Em: BP 690/50) |
| Cameras | ZEISS Axiocam 305 – color CMOS camera; 3,45 µm pixel ZEISS Axiocam 512 mono – monochromatic CCD camera; 3,1 µm pixel |
| Additional equipment | ApoTome.2 for optical sectioning |
| Software | ZEN Blue 2.3 |
| Location | room no. 0.175 (Blue) |
| Phone | ext. 2750 |
| Booking | Calpendo (“Apotome”) |
Z.1 light sheet fluorescence microscope allows fast, gentle multi-view imaging of large specimens. The microscope enables to record the development of living specimens and to deliver exceptionally high information content which includes the inner structures. To image deep into fixed non-transparent biological samples, it is necessary to apply a tissue clearing protocol (e.g. Scale, FocusClear, CUBIC).
| Methods | Fluorescence imaging with lightsheet optical sectioning Dual-site & multi-field scanning, processing at ZEN Black software Time-lapse experiments Multi-positions experiment |
| Illumination | 405nm Solid-state laser, 50mW 445nm Solid-state laser, 25mW 488nm Solid-state laser, 30mW 561nm Solid-state laser, 20mW 638nm Solid-state laser, 75mW Infrared LED light source for trasmitted light |
| Ilumination objectives | Objective Lightsheet Z.1 5x/0.1 Objective Lightsheet Z.1 10x/0.2 |
| Detection objectives | Dry objective Lightsheet Z.1 5x/0.16 Immersion objective Lightsheet Z.1 10x/0.5 Immersion objective Lightsheet Z.1 20x/1.0 Immersion objective Lightsheet Z.1 40x/1.0 Objective Clr Plan-Apochromat 20x/1.0 Corr nd=1,38 (suited for clearing media with RI 1.38, e.g. Scale) Objective Clr Plan-Neofluar 20x/1.0 Corr nd=1.45 (suited for clearing media with RI 1.45, e.g. FocusClear) |
| Laser blocking filters | 405/488/561 tripple excitation dichroic 405/488/561/640 quad excitation dichroic 405/488/640 tripple excitation dichroic 445/561 dual excitation dichroic |
| Emission filter sets | DAPI-GFP (SBS LP 490; Em: BP 420-470 + BP 505-545) DAPI-Cy3 (SBS LP 510; Em: BP 420-470 + BP 575-615) GFP-Cy3 (SBS LP 560; Em: BP 505-545 + BP 575-615) GFP-mCherry (SBS LP 560; Em: BP 505-545 + LP 585) GFP-DRAQ5 (SBS LP 560; Em: BP 505-545 + LP 660) GFP narrow-mCherry (SBS LP 560; Em: BP 505-530 + LP 585) |
| Cameras | camera PCO.Edge 5.5 (sCMOS) – 2 pieces; 6,5 µm pixel |
| Additional equipment | incubation unit (CO2, temperature, humidity) antivibration table for SPIM microscopy PC for storage and data analysis stereomicroscope for sample preparation and system maintenance (Stemi 305 EDU Microscope Set) |
| Software | ZEN Black system 2014 for Lightsheet Z.1 |
| Location | room no. 0.173 (Yellow) |
| Phone | ext. 2750 |
| Booking | Calpendo (“Z1”) |
Leica DMI4000 B – Inverted widefield fluorescence microscope with manual ZX stage, manual objective Z drive and manual objective turret. This microscope is for routine simple applications only. NOT RECOMMENDED FOR IMAGE ACQUISITION. ACQUISITION FEATURES ARE NOT FULLY SUPPORTED.
| Methods | Fluorescence and transmitted light observation (imaging is NOT supported) |
| Illumination | Leica Xenon lamp (75 W) for fluorescence lamp for transmitted light |
| Objectives | N PLAN 10x/0.25 DRY; FWD 17.6 N PLAN L 40x/0.55 CORR DRY; FWD 3.3; CG 0-2 PL APO 63x/1.40 oil |
| Fluorescence filtercubes | CFP (Ex: 436/20; DM 455; Em: 480/40) GFP (Ex: 470/40; DM 500; Em: 525/50) YFP (Ex: 500/20; DM 520; Em: 535/30) RFP (Ex: 546/12; DM 560; Em: 605/75) Cy5 (Y5) (Ex: 620/60; DM 660; Em: 700/75) |
| Cameras | Leica DC350FX – monochromatic CCD camera; 6,4 µm pixel |
| Software | LAS 2.2.0 32bit |
| Location | room no. 3.44 |
| Phone | ext. 3106 |
| Booking | reservation not needed |
The inverted microscope DMi8 with laser scanning confocal head Leica TCS SP8 has differential interference contrast available for all objectives. The confocal head is equipped with two fluorescence PMT detectors and two highly sensitive HyD detectors with time resolved gating function. System is equipped with STED 3X module and 660 nm continual and 775 pulse depletion lasers, and enables super-resolution imaging by stimulated depletion of emission method (STED) with up to 35 nm lateral, 130nm axial resolution.
| Methods | Confocal scanning fluorescence imaging with STED superresolution module Transmitted light imaging Brightfield Time-lapse experiments Multi-positions experiments |
| Illumination | lamp for transmitted light Leica EL6000 with HXP 120W/45C VIS Hg lamp – for fluorescence excitation lasers: – pulsed white light laser (WLL2) – 470-640 nm with 1nm step, 8 parallel laser lines, 1.5 mW per each – continuouse 405 nm DMOD Flexible laser – 50 mW depletion lasers: – 660 nm CW STED, >1.5 W 775 nm pulse STED, >1.5 W |
| Objectives | HC PL APO 10x/0.40 DRY CS; FWD 2.2; CG 0.17 | BF, POL, DIC HC PL APO 100x/1.4 OIL STED WHITE CS2; FWD 0.13; CG 0.17 | BF, POL, DIC |
| Eyepiece filtercubes | DAPI (A) (Ex: 360/40; DM 400; Em: LP 425) FITC (I3) (Ex: 470/40; DM 510; Em: LP 515) RHOD (N2.1) (Ex: 537/45; DM 580; Em: LP 590) Cy5 (Y5) (Ex: 620/60; DM 660; Em: 700/75) |
| Detectors | 2x photomultiplier tube (PMT) 400-800 nm 2x supersensitive hybrid detectors (HyD) 400-720 nm 1x transmitted light detector |
| Confocal head | acousto-optical tunable filter (AOTF) acousto-optical beam splitter (AOBS) standard scanner (1-1800 Hz line frequency) maximum scanner resolution 8192×8192 pixels spectrally tunable detection hardware zoom 0.75x-48x |
| Stage | motorized microscope stage with Super Z-galvo scanning insert for fast and precise Z movement HW autofocus control |
| Additional equipment | antivibration table incubation chamber Okolab (CO2, temperature, humidity) |
| Software | LAS X 64bit – with LAS AF SP8 Dye Finder – 3D visualization, deconvolution and colocalization module |
| Location | room no. 0.174 (Red) |
| Phone | ext. 2433 |
| Booking | Calpendo (“STED”) |
The DeltaVision OMX imaging platform is an advanced multi-mode, super-resolution inverted microscope system which offers super-resolution imaging using 3D Structured Illumination (3D-SIM) and Dense Stochastic Sampling Imaging (DSSI) – Localization Microscopy. The Blaze SIM Module offers dynamic high speed 3D-SIM suitable for live cell super-resolution imaging. Blaze incorporates a proprietary, ultra-fast, structured illumination module, advanced optical platform design and the latest high-speed camera technologies. The system can be used also for super-fast widefield acquisition and photo-kinetic experiments (FRAP).
Methods
Illumination
lamp for transmitted light
InsightSSI (Lumencore) illumination module – for WF imaging; standard shutters, open/close time ~ 1.8 ms excitation lasers – for 3D SIM, TIRF and localization; high speed galvanometer controlled shutters, open/close time ~ 200 μs
3D-SIM resolution at different wavelengths
| Laser | Type | Power (mW) | Expected XY resolution | Expected Z resolution |
| 405 nm | diode | 100 ± 10 | 110 ± 5 nm | 340 ± 10 nm |
| 445 nm | diode | 100 ± 5 | 115 ± 5 nm | 340 ± 10 nm |
| 488 nm | OPSL | 100 ± 4 | 120 ± 5 nm | 340 ± 10 nm |
| 514 nm | OPSL | 100 ± 6 | 120 ± 5 nm | 350 ± 10 nm |
| 568 nm | OPSL | 100 ± 4 | 135 ± 5 nm | 350 ± 10 nm |
| 642 nm | diode | 110 ± 10 | 160 ± 5 nm | 380 ± 20 nm |
Objectives
PLAN APO N 60x/1.42 OIL; FWD 0.15; CG 0.17 | BF, POL, DIC (optimized)
U APO N 60x/1.49 CORR OIL TIRF; FWD 0.1; CG 0.13-0.19 & 23/37°C | BF, POL, DIC
U APO N 100x/1.49 CORR OIL TIRF; FWD 0.1; CG 0.13-0.19 & 23/37°C | BF, POL, DIC
U PLAN S APO 60x/1.3 CORR SILICONE; FWD 0.3; CG 0.15-0.19 & 23/37°C | BF, POL, DIC
InsightSSI excitation filters (for widefield applications only)
DAPI 395.5/29
FITC 477/32
mCherry/Alexa Fluor 568 571/19
Cy5 645.5/15
CFP 438/24
YFP 512.5/15
Emission filters
DAPI 435.5/31
FITC 528/48
mCherry/Alexa Fluor 568 609/37
Cy5 683/40
CFP 477.5/35
YFP 541/22
Cameras
4x pco.edge 5.5 sCMOS, Readout speeds 95 MHz, 286 Mhz, 15 bit, pixel size: 6.5 μm camera, 80 nm 60x, 48 nm 100x
Stage
XYZ nanomover and fast Z-piezo
Additional equipment
antivibration table
sample holders:
Software
OMX Acquisition control software running on OMX Master computer under Windows 7
SoftWoRx – image reconstruction, deconvolution and analysis running on the SI workstation running under the CentOS v6 (Linux)
Dense Stochastic Sampling Imaging (DSSI) algorithm for localization microscopy images
Location: room no. 0.174 (Red)
Phone: ext. 2433
Booking: Calpendo (“OMX”)
The Elyra 7 is a motorized inverted microscope equipped with a super-resolution system that uses a specialized lattice illumination pattern for 3D structured illumination (SIM) images and single-molecule localization methods (SMLM) like STORM and PALM. The lattice illumination allows for faster, gentler, and deeper imaging of live samples, and can reach up to 255 FPS during time-lapse acquisition. The unit has an incubation chamber for live imaging with controlled CO2 and temperature. The signal detection is assured by 2 sCMOS cameras which enable simultaneous detection of two different fluorophores.
| Methods | Lattice SIM, SIM2 Single Molecule Localization Microscopy (3D PALM / dSTORM) TIRF Apotome (optical sectioning) mode Transmitted light imaging Brightfield and Nomarski contrast (DIC) (automatic) Time-lapse experiments (live-cell imaging) Multi-positions experiments |
| Illumination | Halogen 12 V/100W lamp – for transmitted light Metal halide HXP 120 V lamp – for fluorescence Excitation lasers (for TIRF, SIM, SMLM): – 405 nm diode, 50 mW – 488 nm OPSL, 500 mW – 561 nm OPSL, 500 mW – 642 nm OPSL, 500 mW |
| Objectives | EC Plan-Neofluar 10x/0.3 M27 DRY | BF, POL C-Apochromat 40x/1.2 Water Corr M27 | BF C-Apochromat 63x/1.2 Water Corr M27 | BF, DIC α Plan-Apochromat 63x/1.46 Oil Corr M27 | BF, TIRF α Plan-Apochromat 100x/1,46 Oil DIC M27 | BF, DIC, TIRF |
| Eyepiece filtercubes | Laser Blocking Filter for 405/488/561/642 named: – DAPI – FITC – mCherry – Cy5 Simultaneous two-channel shooting: – GFP + mCherry – GFP + Cy5 Sequential four-channel shooting: – DAPI and mCherry + GFP and Cy5 – DAPI and GFP + mCherry and Cy5 |
| Cameras | DuoLink for a parallel imaging on two cameras: – 2x pco.edge sCMOS (version 4.2 CL HS) – thermal stabilization at 0°C / +5 °C / +7 °C – 16-bit depth – high resolution: 2048 x 2048 px – pixel size 6.5 x 6.5 µm – quantum efficiency up to 82% – exposure times from 100 µs to 20 s – maximum frame rate 100 fps |
| Superresolution modules | Lattice SIM, SIM2 Single Molecule Localization Microscopy (3D PALM / dSTORM) |
| Stage | XY Scanning Stage Piezo 130 x 100 (D): – Movement range 130 mm x 100 mm (adjustable) – Max speed 100 mm/s – 0.2 μm resolution – Repeatability +/- 1 μm – Absolute accuracy +/- 5 μm Z-piezo insert: – Movement range at least 100 μm – < 5 nm resolution – Suitable for: Standard microscopic slip: 76 x 26 mm, Petri dish 30 – 35 mm in diameter, LabTek II chamber, 55 x 24 mm |
| Additional equipment | Antivibration table 120 x 90 cm Definite Focus – holding focus to compensate Z drift Dark incubation unit (CO2, temperature < 40°C, humidity) – Imaging chamber for Z-piezo inserts PC for storage and data analysis |
| Software | ZEN 3.0 (black edition) 64-bit Modules: – Lattice SIM2/SIM2 Apotome – SMLM (PALM/dSTORM) module – 3D-PALM – Measurement – Multi Channel – Panorama – Manual Extended Focus – Image Analysis – Time Lapse – Tiles & Positions – Z-stack – Extended Focus – Autofocus – Colocalisation – Connect Entry (ZEN blue edition) |
| Location | room no. 0.174 (Red) |
| Phone | ext. 2433 |
| Booking | Calpendo (“Elyra 7”) |
Q-PHASE is a unique instrument for quantitative phase imaging (QPI). It is based on a robust and fully motorized inverted transmission microscope platform. Multiple imaging modes are integrated such as fluorescence, simulated DIC, high-pass filter and brightfield. It is a perfect tool for long-term label-free imaging of living cells. Sophisticated user-friendly software is available for microscope control, image acquisition and subsequent cell segmentation and highly advanced data mining and analysis.
| Methods | Quantitative phase imaging in TL mode Additional fluorescence imaging Time-lapse experiments Multi-position experiments QPI image advanced analysis |
| Illumination | Lumencore Spectra X – solid-state light source for fluorescence Transmitted light source halogen lamp EKE 21V |
| Objectives | Nikon PLAN ACHROMAT 4x/0.1 DRY; FWD 30 | BF, QPhase, Hologram, POL, DIC, High Pass Filter (PH) Nikon PLAN FLUOR 10x/0.3 DRY; FWD 16 | BF, QPhase, Hologram, POL, DIC, High Pass Filter (PH) Nikon PLAN FLUOR 20x/0.5 DRY; FWD 2.1 | BF, QPhase, Hologram, POL, DIC, High Pass Filter (PH) Nikon PLAN APO 40x/0.95 DRY, FWD 0.17-0.25 | BF, QPhase, Hologram, POL, DIC, High Pass Filter (PH) Nikon PLAN APO 60x/1.4 OIL, FWD 0.13| BF, QPhase, Hologram, POL, DIC, High Pass Filter (PH) |
| Filter set | DAPI FITC/Texas Red |
| Spectra X excitation filters | DAPI 390/22 CFP 434/21 GFP 470/24 FITC 494/20 YFP 511/16 DsRED 555/28 Texas Red 575/25 |
| Dichronic mirrors | QPl-Fluorescence Beam Combiner SP 635 (limits detection of emission wavelengths below 635 nm) 436/514/604 nm tripple band dichroic DAPI/FITC/Texas Red (transmission bands Tavg > 90%: 446-468, 520-540, 614-642) 440/520 nm dual band dichroic CFP/YFP (transmission bands Tavg > 90%: 449-483, 530-569) 493/574 nm dual band dichroic GFP/DsRed (transmission bands Tavg > 90%: 500-529, 584-679) |
| Emission filter wheel | DAPI 447/60 CFP 472/30 GFP 514/30 FITC 531/22 YFP 542/27 DsRed 617/73 Texas Red 624/40 |
| Cameras | XIMEA MR4021MC – monochromatic CCD camera for TL; 7,4 µm pixel Andor Zyla 5.5 – monochromatic sCMOS camera for fluorescence; 6,5 µm pixel |
| Aditional equipment | incubation unit (CO2, temperature, humidity) |
| Software | Q-PHASE software |
| Location | room no. 0.175 (Blue) |
| Phone | ext. 2750 |
| Booking | Calpendo (“QPI”) |
