Confocal microscopy
Equipment
- Equipment
Zeiss LSM 780
Motorized Zeiss Axio Observer.Z1 inverted microscope.
Technical Information and capabilities
Objectives
Magnification | Objective type | NA | Working distance (mm) | phase contrast | Immersion |
---|---|---|---|---|---|
10x | Plan-Apochromat | 0.45 | 2.1 | - | Dry |
20x | Plan-Apochromat | 0.80 | 0.55 | - | Dry |
25X | LD LCI Plan- | 0.8 | 0.57 | - | Water, Oil, Glyc |
40x | Plan-Neofluor | 1.30 | 0.20 | - | Oil |
40X | C-Apochromat (on request) | 1.2 | 0.28 | - | Water |
63x | Plan-Apochromat | 1.40 | 0.19 | - | Oil |
Laser lines
Laser | Wavelength (nm) | Example of fluorochromes |
---|---|---|
Diode | 405 | DAPI, Hoechst |
Argon | 458 | CFP |
Argon | 488 | FITC, Alexa Fluor 488, GFP, Cy2, SytoxGreen, Acr. Orange/DNA |
Argon | 514 | YFP, Cy3 |
Diode | 561 | PI, TRIT/Rhodamine, Mitotracker Orange, Alexa Fluor 546, DsRed2, Alexa Fluor 594, Texas Red, HcRed1 |
Helium-Neon | 633 | Alexa 633, Cy5, APC, Alexa Fluor660, Alexa Fluor 647 |
Filters
Fluorescence filters for VIS-mode
Zeiss filter set 49 | Zeiss filter set 38 | Zeiss filter set 43 |
---|---|---|
DAPI/ Hoechst | FITC/Alexa Fluor 488/ GFP | TRITC/ Cy3/ Alexa Fluor 546 |
Transmitted light in Bright-field and DIC (ask for the DIC prisms if you need them).
Detectors
- 2 PMT detectors (One with extra cooling for increased sensitivity in infrared)
- GaAsP spectral detector
- Transmitted light detector (T-PMT)
Software and accessories for live cell and multi-well imaging
- Zeis ZEN 2010 software. Physiology
- FCS for GaAsP
- ROI HDR
- Image Correlation Spectroscopy analysis packages for ZEN 2010
- Definite Focus hardware for auto-focus system
- Heating Insert P S1 (temperature control)
- Incubator PM S1 (Temperature and CO2 control)
- 40x and 63x water immersion objectives are available upon request
System Changes
- Computer main drive was upgraded to a new bigger SSD (6.3.2024)
Keywords
Inverted microscope | Multi-well | Confocal | Incubator | Environmental chamber | Definite focus | Water immersion |
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Zeiss LSM 880
Zeiss inverted confocal microscope with GaAsP detector, AiryScan (Fast) detector, PicoQuant FLIM detector and a full environmental chamber.
Technical Information and capabilities
Objectives
Magnification | Objective type | NA | Working distance (mm) | phase contrast | Immersion |
---|---|---|---|---|---|
10x | Plan-Apochromat | 0.45 | 2 | - | Dry |
20x | Plan-Apochromat | 0.80 | 0.55 | - | Dry |
40x | C-Apochromat | 1.20 | 0.28 | - | Water |
40x | Plan-Apochromat | 1.40 | 0.13 | - | Oil |
63x | Plan-Apochromat | 1.40 | 0.19 | - | Oil |
Laser lines
Laser | Wavelength (nm) | Example of fluorochromes |
---|---|---|
Diode | 405 | DAPI, Hoechst |
Argon | 458 | CFP |
Argon | 488 | FITC, Alexa Fluor 488, GFP, Cy2, SytoxGreen, Acr.Orange/DNA |
Argon | 514 | YFP, Cy3 |
Diode | 561 | PI, TRITC/Rhodamine, Mitotracker Orange, Alexa Fluor 546, DsRed2, Alexa Fluor 594, Texas Red, HcRed1 |
Helium-Neon | 633 | Alexa 633, Cy5, APC, Alexa 660, Alexa 647 |
AiryScan Filters
- Plate
- BP 420-480 + BP 495-550
- BP 420-480 + BP 495-620
- BP 420-480 + LP 605
- BP 465-505 + LP 525
- BP 495-550 +LP 570
- BP 570-620 + LP 645
Main Beam Splitters
- MBS 458
- MBS 458/514
- MBS 458/461
- MBS 488
- MBS 488/561
- MBS 488/561/633
- MBS T89/R20
- Plate
Secondary Beam Splitters
- Mirror
- SBS LP 490
- SBS LP 515
- SBS LP 545
- SBS LP 635
- SBS SP 545
- SBS SP 610
- Plate
Fluorescence filters for VIS-mode
Zeiss filter set 49 | Zeiss filter set 38 | Zeiss filter set 43 | Zeiss filter set 50 |
---|---|---|---|
DAPI/ Hoechst | FITC/ Alexa Fluor 488/ GFP | TRITC/ Cy3/ Alexa Fluor 546 | Cy5, Alexa Fluor 647 |
Transmitted light in Bright-field and DIC (ask for the DIC prisms if you need them).
Detectors
- 2 PMT detectors (One with extra cooling for increased sensitivity in infrared)
- 1 GaAsP detector
- Airyscan detector
- Transmitted light detector (T-PMT)
Software and accessories for live cell and multi-well imaging
- Zeis ZEN 2
- Modules: Experiment designer, Tiles & Positions, FRAP and FRET
- Definite Focus hardware autofocus system
- Scanning stage for multipoint or mosaic imaging
- Large environmental chamber
- Incubator insert PM S1 (Temperature and CO2 control): Observation area 32 x 30 mm (oval) https://www.pecon.biz/products/product/?pid=5271%7CHeating_Insert_P_S
- 40x and 63x water immersion objectives are available upon request
Environmental Chamber Temperature Setting
The readout of the temperature probe differs slightly from the media temperature in the sample chamber. To heat the sample to +37 C, adjust the Incubator to:
- H Insert P +38.0 °C
- H Unit XL +38.2 °C
Test your own fluorophores on this optical system
To do so, click on the link below, choose the right configuration and add your own fluorophore. You will see the efficiency of the system for that specific label.
https://www.fpbase.org/microscope/fGbZLzo4KpjDLnCZqK9m6G/
Keywords
Inverted microscope | Multi-well | Confocal/ multiphoton | Super-resolution | Incubator | Large environmental chamber | Definite focus | Water immersion |
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Zeiss LSM 7 MP
Upright multiphoton microscope optimized for intravital imaging of anaesthetized small laboratory animals.
The system allows acute as well chronic in vivo imaging experiments on internal (e.g. brain, kidney, liver etc.) and external organs (e.g. skin and eye).
The system can also be used for imaging fixed thick tissue samples.
Technical Information and capabilities
Objectives
Magnification | Objective type | NA | Work distance (mm) | Immersion | Coverglass thickness (mm) |
---|---|---|---|---|---|
20x | W Plan-Apochromat | 1.0 | 1.8 | Water | 0 |
32x | C-Achroplan | 0.85 | 0.85 | Water | 0-0.17 |
Lasers
- Coherent Chameleon Vision II, tunable wavelength: 690-1010nm
- APE Chameleon Compact OPO (optical parametric oscillator), tunable wavelength 1050-1300nm
Emission filters
- Reflected PMT's: BP 500-550 (green), BP 565-610 (red)
- Transmitted PMT's: BP 500-550 (green), BP 565-610 (red)
- Transmitted light filters for Bright-field and by request DoDt contrast (inclined illumination)
Light Sources
- Widefield fluorescence and reflected light
- HXP 120 C metal halide lamp (fluorescence)
- LED-light source F1 (general-purpose light source)
Detectors
- 2 reflected light non-descanned PMT detectors with the possibility for filters
- 1 optional reflected light non-descanned nosepiece GaAsP detector without filters
- 2 transmitted light non-descanned PMT detectors with emission filters available on request
- 1 transmitted light non-descanned PMT detector available on request
Software and accessories for live cell and multi-well imaging
- ZEN 2011 software
- LinLab software for Scientifica motorized stage
- Dark microscope enclosure
- Univentor gas anesthesia system with mask, induction boxes and scavenger
- PhysioSuite hear/breath is an oxygenation monitoring system for mice and rats
- Animal's heating pad at the microscope stage to keep the animal warm during the anaesthesia
- Scientifica adjustable motorized microscope stage with a variety of intravital animal fixation adapters
- LED-light source F1 with flexible light guides for manipulations under the microscope
- Prior motorized microscope stage with multi-point scanning option available on request
- Dark environmental chamber (incubator XL) for minimization of stray light available on request
- Heating unit for temperature control of environmental chamber available on request.
Keywords
Upright microscope | Multi-well | Wide-field | Confocal/ multiphoton | Intravital imaging | In vivo imaging | Animal's heating pad | Dark environmental chamber | gas anesthesia |
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Leica SP8 CARS
Leica inverted multi-photon microscope with two HyD detectors, resonant scanner, tunable CARS laser and a full environmental chamber.
Technical Information and capabilities
Objectives
Magnification | Objective type | NA | Working distance (µm) | Immersion | Coverglass thickness (mm) | XY res. (nm) | Z res. (nm) |
---|---|---|---|---|---|---|---|
10x | HC PL APO CS | 0.40 | 360 | Multi | - | 488 | 4093 |
25x | HCX IR APO L | 0.95 | 2400 | Water | 0.17 | 205 | 550 |
20x | HC PL APO CS2 | 0.75 | 680 | Multi | - | 260 | 1108 |
40x | HC PL APO CS2 | 1.10 | 650 | Water | 0.14-0.18 | 177 | 377 |
63x | HC PL APO CS2 | 1.20 | 300 | Water | 0.14-0.18 | 163 | 290 |
63x | HC PL APO CS2 | 1.30 | 300 | Glycerol | 0.14-0.19 | 150 | 276 |
Single-photon laser lines
Laser | Wavelength (nm) | Example of fluorochromes |
---|---|---|
Diode (Violet) | 405 | DAPI, Hoechst |
Argon (Blue) | 458 | CFP |
Argon (Blue) | 476 | - |
Argon (Blue) | 488 | FITC, Alexa Fluor 488, GFP, Cy2, SytoxGreen, Acr.Orange/DNA |
Argon (Blue) | 496 | - |
Argon (Blue) | 514 | YFP, Cy3 |
DPSS (Lime) | 561 | PI, TRITC/Rhodamine, Mitotracker Orange, Alexa Fluor 546, DsRed2, Alexa Fluor 594, Texas Red, HcRed1 |
Helium-Neon (Orange) | 594 | - |
Helium-Neon (Red) | 633 | Alexa 633, Cy5, APC, Alexa 660, Alexa 647 |
Multiphoton laser (CARS, TPEF, SHG)
- APE picoEmerald
- Wavelengths: 1064.5 nm and tunable 780-940 nm
Detectors
- 2 high sensitivity hybrid detectors (HyD)
- 1 PMT detector
- 2 reflected light non-descanned PMT detectors with the possibility for filters
- 2 transmitted light non-descanned PMT detectors with the possibility for filters
Software and accessories for live cell and multi-well imaging
- Full-enclosure type temperature-controlled environmental chamber for live-cell imaging and bleaching FRAP applications.
- An inverted temperature-controlled slow-flow imaging chamber available on request
Keywords
Inverted microscope | Multi-well | CARS | Confocal/multi-photon | Live cell imaging | Environmental chamber | Water immersion |
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Leica TCS SP8 X
Leica TCS SP8 X with white light laser. Laser scanning confocal microscope for both fixed and live-cell imaging. Mounted on a motorized Leica inverted microscope.
Technical Information and capabilities
Objectives
Magnification | Objective type | NA | Working distance (µm) | Immersion | Coverglass thickness (mm) | XY res. (nm) | Z res. (nm) |
---|---|---|---|---|---|---|---|
10x | HC PL APO CS2 | 0.40 | 2740 | Dry | 0.17 | 488 | 2630 |
20x | HC PL APO CS2 | 0.75 | 620 | Dry | 0.17 | 260 | 649 |
40x | HC PL APO CS2 | 1.30 | 240 | Oil | 0.17 | 150 | 299 |
63x | HC PL APO CS2 | 1.40 | 140 | Oil | 0.17 | 139 | 236 |
63x | HC PL APO CS2 | 1.20 | 300 | Water | 0.14-0.18 | 163 | 290 |
Lasers
- 405 nm diode
- Supercontinuum laser, freely tunable between 470-670 nm, maximum of 8 simultaneous laser lines
Detectors, Software and accessories for live cell and multi-well imaging
- 2 high sensitivity hybrid detectors (HyD)
- 2 PMT detectors
- 1 transmitted light detector (T-PMT)
The SP8 X has a full-enclosure type temperature-controlled environmental chamber for imaging live cells. While the chamber and microscope are heating up or cooling down the light path distances are constantly changing. Because of this it is not recommended to image while the chamber temperature is changing. To keep the chamber at a steady temperature, it should be either heated (+37 °C) or at room temperature for the whole day. The first reservation created for a given day determines the temperature of the system.
Keywords
Inverted microscope | Multi-well | Confocal | Live cell imaging | Environmental chamber | White light laser |
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Leica Stellaris 8 FALCON / DLS
Leica Stellaris 8 inverted confocal microscope with 5 sensitive HyD detectors and tunable white-light laser excitation source.
Highlights:
- FALCON module for fast fluorescence lifetime measurement
- Digital Light-Sheet module
- TauSense detection
- Fast resonant scanner
- LASX Navigator
- Leica Lightning
- NIR excitation (up to 790nm) and detection (up to 850nm)
The LMU have made a nice video covering the capabilities of the Stellaris 8 and its FALCON module (not DLS). Definitely worth a look.
Technical information:
Microscope
- Leica DMi8 inverted microscope
- Motorized scanning stage
- AFC hardware autofocus
- OKOlab full-enclosure incubator with temperature, CO2, and humidity control
Objectives
Magnification | Objective type | NA | Working distance (µm) | Immersion | Coverglass thickness (mm) |
---|---|---|---|---|---|
10x | HC PL APO CS2 | 0.40 | 2740 | Dry | 0.17 |
20x | HC PL APO CS2 | 0.75 | 620 | Dry | 0.17 |
40x | HC PL APO CS2 CORR | 1.25 | 350 | Glycerol | 0.14-0.18 |
63x | HC PL APO CS2 | 1.40 | 140 | Oil | 0.17 |
63x | HC PL APO CS2 motCORR | 1.20 | 300 | Water | 0.14-0.18 |
Lasers
- 405 nm diode laser
- 488 nm laser for FRAP
- Supercontinuum laser, freely tunable between 440-790nm, maximum of 8 simultaneous laser lines
Scanner
Tandem scanner with both traditional point scanner and fast 8kHz resonant scanner.
Five sensitive HyD detectors:
- 2 HyD S - all round detector for high performance detection throughout the spectrum
- 2 HyD X - specialized for fast fluorescence lifetime imaging in the visible to the far-red
- 1 HyD R - specialized for detection above 740nm (incl. fast fluorescence lifetime)
Please note that all 5 detectors are suitable for regular confocal imaging. However, the HyD X's and HyD R are not tolerant of over exposure. If overexposed they will shut down and may be permanently damaged. You can find out more about the Leica HyD detectors and their capabilities here.
Software Leica LAS X
4.5.04.6.0 (5.10.2023)- 4.7
Andor Dragonfly
Technical Information and capabilities
Andor Dragonfly 505
Objectives
Magnification | Objective type | NA | Working distance (mm) | Phase-contrast | Immersion | PFS |
---|---|---|---|---|---|---|
10x | Plan Fluor DL | 0.30 | 15.2 | Ph1 | Dry | + |
20x | Plan Apo λ | 0.75 | 1.0 | - | Dry | + |
40x 1), 2) | Apo LWD | 1.15 | 0.6 | - | Water | + |
60x | Plan Apo VC | 1.2 | 0.29 | - | Water | + |
100x 1) | SR Apo TIRF | 1.49 | 0.12 | - | Oil | + |
40x | Plan Apo λ | 1.25 | 0.3 | - | Silicone oil | + |
1) Not Plan corrected
2) Suitable for NIR labels. Chromatic correction 405-950nm
Lasers
- 404nm, 100mW
- 488nm, 150mW
- 561nm, 100mW
- 642nm, 110mW
- 730nm, 30mW
Filters
For visual inspection of your sample through the eyepiece
- DAPI-50606
- GFP-30309
- TRITC-B 05
- mCherry-C
For imaging
- 445 -46 nm
- 521 -38 nm
- 594 -43 nm
- 685 -47nm
- 809 -90nm
- TR-POL-DIC-FWL, Polarizer
- 445/521/594/685/809
Cameras
- Andor Zyla 4.2 sCMOS
- Andor iXon 888 U3 EMCCD
Software
- Fusion 2.0
- ClearView-GPU online deconvolution
- iXon SRRF-Stream real-time super-resolution
- Bitplane Imaris
Test your own fluorophores on this optical system
To do so, click on the link below, choose the right configuration and add your own fluorophore. You will see the efficiency of the system for that specific label.
https://www.fpbase.org/microscope/MHoNXQQFSXUuAowQ34PYg8/?d=andor-ixon-life-888-bv
Data Transfers From the Dragonfly Computer
If you're exporting or transferring data from the Dragonfly computer and not using the microscope, please start the Fusion OFFLINE version from the desktop. Using the regular Fusion while the microscope is off will confuse it and cause problems to the next user.