iMIC TIRF FD-FLIM and TGL
The microscope is custom build around the iMIC stand (Till Photonics/FEI) with H101 motorized stage (Prior).
The iMIC stand is equipped with:
Objective carousel (up to 4 Olympus objectives). Available objectives: UPLFLN 4x (WD 17, NA 0.13), LUCPLFLN 20x (WD 6.6-7.8, NA 0.45) and 60x (WD 1.5-2.2, NA 0.7); Super Apochromats UPLSAPO 4x (NA 0.13), 10x (NA 0.4), 20xo (NA 0.85), 40x (NA 0.95), 60xw (NA 1.2), 100xo (NA 1.4) and TIRF APON 60x (NA 1.49).
Focus drive (combines a piezo-element having 250 µm of fine travel (resolution 50 nm) with 25 mm of coarse movement).
Motorized beamsplitter changer (for up to 3 octo-cube beamsplitters).
Dichrotome Level A - Dual emission Imaging Extension – single beam splitter separate the emission light into two independent color/polarization channels and each channel is simultaneously projected onto half of the CCD-chip.
Dichrotome Level B – extend the Dichrotope Level A with possibility to additionally split the emission light into two independent channels using Polarizing Beam splitter (for wavelength 400-700 nm) and each channel could be simultaneously projected onto own CCD-chip. For such type applications two identical CCD Clara cameras (Andor) available upon request (default position of those cameras is on STED-FLCS microscope).
TuCam (Andor) – two camera adapter that in default configuration coupled to one output port of Dichrotome Level B. Used for switching between two cameras: EMCCD 885 (Andor) – used for single molecule experiments and ICCD Li2CAM (Lambert Instruments) – used for FLIM/TGL.
The Prior stage is equipped with adapters for a wide range of sample holders: multiwell plates, slides, petri dishes, Chamlide magnetic multiwell chambers (available WL-2460-10 and CM-S22-4) and closed Flow Cell Systems (Focht Chamber System FCS2 from Bioptech and Magnetic closed chamber CF-T12 from Chamlide).
For the illumination there are different options available.
The iMIC stand is equipped with:
Objective carousel (up to 4 Olympus objectives). Available objectives: UPLFLN 4x (WD 17, NA 0.13), LUCPLFLN 20x (WD 6.6-7.8, NA 0.45) and 60x (WD 1.5-2.2, NA 0.7); Super Apochromats UPLSAPO 4x (NA 0.13), 10x (NA 0.4), 20xo (NA 0.85), 40x (NA 0.95), 60xw (NA 1.2), 100xo (NA 1.4) and TIRF APON 60x (NA 1.49).
Focus drive (combines a piezo-element having 250 µm of fine travel (resolution 50 nm) with 25 mm of coarse movement).
Motorized beamsplitter changer (for up to 3 octo-cube beamsplitters).
Dichrotome Level A - Dual emission Imaging Extension – single beam splitter separate the emission light into two independent color/polarization channels and each channel is simultaneously projected onto half of the CCD-chip.
Dichrotome Level B – extend the Dichrotope Level A with possibility to additionally split the emission light into two independent channels using Polarizing Beam splitter (for wavelength 400-700 nm) and each channel could be simultaneously projected onto own CCD-chip. For such type applications two identical CCD Clara cameras (Andor) available upon request (default position of those cameras is on STED-FLCS microscope).
TuCam (Andor) – two camera adapter that in default configuration coupled to one output port of Dichrotome Level B. Used for switching between two cameras: EMCCD 885 (Andor) – used for single molecule experiments and ICCD Li2CAM (Lambert Instruments) – used for FLIM/TGL.
The Prior stage is equipped with adapters for a wide range of sample holders: multiwell plates, slides, petri dishes, Chamlide magnetic multiwell chambers (available WL-2460-10 and CM-S22-4) and closed Flow Cell Systems (Focht Chamber System FCS2 from Bioptech and Magnetic closed chamber CF-T12 from Chamlide).
For the illumination there are different options available.
- Trans-illumination – Green LED. Supplied through long working distance condenser IX-LWUCD NA 0.55 (Olympus).
- Widefield Epifluorescence illumination:
- High-quality modulating LEDs in Multi-LED excitation source (Lambert Instruments). Digital modulation up to 120 MHz. Integrated into LIFA-X system (Lambert Instruments) for widefield Frequency Domain Fluorescence Lifetime Imaging (FD-FLIM) and widefield Time-Gated Luminescence imaging (TGL). Four lines with peak light intensities at wavelengths 368, 432, 477, 532 nm are available.
- Ultra-fast (< 3ms) filter switching Xenon Flash lamp – Oligochrome(Till Photonics/FEI) with up to 5 excitation filters capacity in the wavelength range from 320 nm to 700 nm. Suitable for high-speed multi-color and ratio widefield imaging. Combined with laser TIRF through Polytrope Imaging-Mode Switch.
- Objective TIRF illumination – MultiLaser/SOLE-6 (Lambert Instruments/Omicron) - High-Speed digital and/or analogue modulation laser light engine fiber coupled through Polytrope and position controlled by Yanus Scan Head. Contains 5 PhoxX laser diodes for CW and modualated applications (lines: 405, 445, 488, 515, 638 nm) and 1 DPSS line modulated by AO-modulator (Cobolt Jive 561 nm). Laser diodes are also integrated into LIFA system for TIRF- FD-FLIM measurements. Polytrope and Yanus combination for all MultiLaser lines provides variable angle with azimuthal scan (2D) TIRF functionality and FRAP and photoactivation possibilities.
iMIC STED-FLCS and FCCS
The microscope is custom build around the iMIC stand (Till Photonics/FEI) with Motorized TILL stage.
The iMIC stand is equipped with:
Objective carousel (up to 4 Olympus objectives). Available objectives: UPLFLN 4x (WD 17, NA 0.13), LUCPLFLN 20x (WD 6.6-7.8, NA 0.45) and 60x (WD 1.5-2.2, NA 0.7); Super Apochromats UPLSAPO 4x (NA 0.13), 10x (NA 0.4), 20xo (NA 0.85), 40x (NA 0.95), 60xw (NA 1.2), 100xo (NA 1.4) and TIRF APON 60x (NA 1.49).
Focus drive (combines a piezo-element having 250 µm of fine travel (resolution 50 nm) with 25 mm of coarse movement).
Motorized beamsplitter changer (for up to 3 octo-cube beamsplitters).
Dichrotome Level A - Dual emission Imaging Extension – single beam splitter separate the emission light into two independent color/polarization channels and each channel is simultaneously projected onto half of the CCD-chip. By default used one of two available Clara CCD cameras (Andor). The second Clara CCD camera is used for STED donut alignment.
The TILL stage is equipped with adapters for a wide range of sample holders: slides, small petri dishes, Chamlide magnetic multiwell chambers (available WL-2460-10 and CM-S22-4) and closed Flow Cell System (Magnetic closed chamber CF-T12 from Chamlide).
For the illumination there are different options available.
Software: IQ 2.7 (Andor), SPCImage 5.2 and Burst analyzer 2.0 (Becker&Hickl), ImageJ/FIJI, Matlab
The iMIC stand is equipped with:
Objective carousel (up to 4 Olympus objectives). Available objectives: UPLFLN 4x (WD 17, NA 0.13), LUCPLFLN 20x (WD 6.6-7.8, NA 0.45) and 60x (WD 1.5-2.2, NA 0.7); Super Apochromats UPLSAPO 4x (NA 0.13), 10x (NA 0.4), 20xo (NA 0.85), 40x (NA 0.95), 60xw (NA 1.2), 100xo (NA 1.4) and TIRF APON 60x (NA 1.49).
Focus drive (combines a piezo-element having 250 µm of fine travel (resolution 50 nm) with 25 mm of coarse movement).
Motorized beamsplitter changer (for up to 3 octo-cube beamsplitters).
Dichrotome Level A - Dual emission Imaging Extension – single beam splitter separate the emission light into two independent color/polarization channels and each channel is simultaneously projected onto half of the CCD-chip. By default used one of two available Clara CCD cameras (Andor). The second Clara CCD camera is used for STED donut alignment.
The TILL stage is equipped with adapters for a wide range of sample holders: slides, small petri dishes, Chamlide magnetic multiwell chambers (available WL-2460-10 and CM-S22-4) and closed Flow Cell System (Magnetic closed chamber CF-T12 from Chamlide).
For the illumination there are different options available.
- Trans-illumination – Green LED. Supplied through long working distance condenser IX-LWUCD NA 0.55 (Olympus).
- Widefield Epifluorescence illumination: Ultra-fast (< 1.6 ms) switching monochromator Xenon Flash lamp – Polychrome V (Till Photonics/FEI) variable wavelength range from 320 nm to 680 nm. Suitable for high-speed multi-color and ratio widefield imaging.
- Confocal Epifluorescence illumination:
- Picosecond Pulsed supercontinuum laser Solea (Picoquant). Spectral range 480-900 nm (variable slit width from 3 till 15 nm). Internal oscillator 2.5 - 40 MHz. Triggered for TCSPC applications (FLIM, FCS) and syncronised with VISIR-765-STED laser for STED-FLCS applications and LLC for FCCS applications. Directly coupled through Yanus Scan Head. Yanus provides Beam Park, line, raster and orbital scan modes.
- CW laser line combiner (LLC) with two lines - Cobolt Calypso Dual 488/532. Lines are selectable with AOTF (AA Optoelectronics). Used as the second line for cross-correlation with Solea laser.
- Picosecond Pulsed laser module VISIR-765-STED. Average power 1.5 W. Syncronised with Solea laser through variable picosecond delayer (MicroPhotonDevices). Used for STED “donut” formation by Vortex Phase Plate.
Software: IQ 2.7 (Andor), SPCImage 5.2 and Burst analyzer 2.0 (Becker&Hickl), ImageJ/FIJI, Matlab
Nikon PALM (under construction)
The microscope is custom build around the Eclipse TE2000-U stand (Nikon) with Motorized Prior stage H117 controlled with ProScan III.
The TE2000-U stand is equipped with:
Objective carousel (up to 6 Nikon objectives). Available objectives: Plan Fluor 10x (NA 0.3), Plan Fluor 40x (NA 0.6) and TIRF APO 100x (NA 1.49).
Remote focus controller.
Manual beamsplitter changer (for up to 6 beamsplitter cubes).
The Prior stage is equipped with adapters for a wide range of sample holders: multiwell plates, slides, petri dishes, Chamlide magnetic multiwell chambers (available WL-2460-10 and CM-S22-4) and closed Flow Cell Systems (Focht Chamber System FCS2 from Bioptech and Magnetic closed chamber CF-T12 from Chamlide).
For the illumination there are different options available.
Software: Nikon Confocal Microscope C1 EZ-C1 Software, MicroManager, LabView, ImageJ/FIJI, Matlab.
The TE2000-U stand is equipped with:
Objective carousel (up to 6 Nikon objectives). Available objectives: Plan Fluor 10x (NA 0.3), Plan Fluor 40x (NA 0.6) and TIRF APO 100x (NA 1.49).
Remote focus controller.
Manual beamsplitter changer (for up to 6 beamsplitter cubes).
The Prior stage is equipped with adapters for a wide range of sample holders: multiwell plates, slides, petri dishes, Chamlide magnetic multiwell chambers (available WL-2460-10 and CM-S22-4) and closed Flow Cell Systems (Focht Chamber System FCS2 from Bioptech and Magnetic closed chamber CF-T12 from Chamlide).
For the illumination there are different options available.
- Trans-illumination – Halogen Lamp.
- Widefield Epifluorescence illumination: Mercury Lamp
- Objective TIRF illumination: Laser diodes: 405 nm and 561 nm. Used for PALM
- Confocal Epifluorescence illumination:Gas lasers Argon (488 nm) and Helium-Neon (543 nm). Gas lasers are fiber coupled to Nikon C1 Confocal System unit
Software: Nikon Confocal Microscope C1 EZ-C1 Software, MicroManager, LabView, ImageJ/FIJI, Matlab.
