ISU Leica SP8 Confocal System/White Light Laser/Falcon
Microscope:
-DMi8 CS inverted microscope; DIC
-Air Table / Compressor
-Super Z Galvo Stage; optional Universal or Multiwell plate inserts
-LMT200-T high precision scanning stage
-Adaptive Focus Control
-Turret Cooling
-LED brightfield light source
-Lumencor SOLA SE II white LED light source for wide-field fluorescence
-Leica DFC7000 T cooled color camera for non-confocal light and fluorescence imaging
-Transmitted light PMT
-Blue/green/red/far red filter cubes for wide-field fluorescence:
–DAPI bandpass filter cube. Excitation: BP 350/50, Emission: BP 460/50
–LED 470 filter cube. Excitation: 470/40, Dichroic: 495, Emission: 525/50
–RHOD bandpass filter cube. Excitation: BP 546/10, Emission: BP 585/40
–Y5 bandpass filter cube. Excitation: BP 620/60, Emission: BP 700/75
OBJECTIVES:
Summary: 5x; 10x; 20x-imm; 40x-oil; 40x-water; 63x-oil; 63x-glyc; 100x-oil.
1) HC PL FLUOTAR 5x/0.15. For use with and without coverglass, no immersion. Free working distance: 12.0 mm.
2) HC PL APO 10x/0.40 CS2. For use with 0.17 mm coverglass. Air immersion. Free working distance: 2.2 mm.
3) HC PL APO 20x/0.75 IMM CORR CS2. Correction collar for use with and without coverglass and use with water, glycerine and oil immersion. Free working distance: 0.67 mm (water immersion and 0.17 mm coverglass).
4) HC PL APO 40x/1.30 Oil CS2. Oil immersion. Free working distance: 0.24 mm.
5) HC PL APO 40x/1.10 W motCORR CS2. Motorized correction collar for adjustment of coverglass thickness from 0.14-0.18 mm, water and saltwater immersion and within the temperature range 20-40°C. Free working distance: 0.65 mm.
6) HC PL APO 63x/1.40 OIL CS2. Oil immersion. Free working distance: 0.14 mm.
7) HC PL APO 63x/1.30 GLYC CORR CS2. Glycerol immersion. Correction collar for use with coverglass (0.14-0.19 mm), glycerol immersion (80% glycerol / 20% water) and within the temperature range 20-40°C. Free working distance: 0.30 mm.
8) HC PL APO 100x/1.40 OIL CS2. Oil immersion. Free working distance: 0.13 mm.
TCS SP8 Confocal Scan Head
-Tandem Scanner
–Non-resonant (conventional/variable) Scanner: 1 – 1800 Hz (7 fps @ 512 x 512 px, 84 fps @ 512 x 16 px), line frequency up to 3600 lines/second (bidirectional), max. scan format 8192 x 8192 px, scan field 22 mm diag.
–Resonant Scanner (ultra-fast, reduced photodamage): 8000 Hz (28 fps @ 512 x 512 px, 290 fps @ 512 x 16 px), line frequency up to 16,000 lines/second (bidirectional), max. scan format 2496 x 2496 px, scan field 13 mm diag. Optical field rotation 200°.
-Notch filter set (provides additional suppression of excitation light) – NF 445; NF 488; NF 514; NF 458/514; NF 488/561/633; NF 594; NF 445/594
-5-channel filter?free prism detector system. Emission bands freely tunable with edge-positioning precision of 1nm. Lambda-Scan mode permits recording of spectral image series.
–Two PMTs (Hamamatsu R 9624 photomultiplier/ 40 MHz sampling/ detection range 400 – 800 nm)
–Three HyD hybrid Spectral detectors. Very low dark noise; Photon counting capability; Enables time gated detection.
-Programmable control panel
Excitation
-Continuous wave lasers
–405 nm Diode Laser (50 mW), DMOD
–Argon Laser (65 mW): 458 nm, 476 nm, 488 nm, 496 nm, 514 nm
-Pulsed lasers
–White light laser WLL2. Select up to 8 lines, individually tunable in steps of 1 nm from 470 to 670 nm, ~1.5 mW power per line. Pulsed, 80 MHz. 8 channel AOTF (Acousto-Optical-Tunable-Filter) for rapid modulation of laser intensity.
–Recording of two dimensional excitation-emission spectra supported by software for setup and display of 2D spectra.
–Lightgate function for avoidance of reflected laser light
–Pulsed 440 nm laser, LDH-P-C-440B. Suited to lifetime based measurements including FLIM. Software controlled selection of pulse frequency (40, 20, 10, and 5 MHz).
-LAS X Lightning Expert software for adaptive deconvolution
-FALCON (FAst Lifetime CONtrast) real time FLIM detection; FLIM-FRET analyzer.
-HPZ840 Workstation with Windows 10 Professional (64 bit) operating system. 2x Intel 4-core Xeon E5-2637 V4 3.5 GHz. 128 GByte RAM. NVIDIA Quadro GP100 16GB Reference GPU (3584 cores).
-Leica LAS X system software; LAS X MicroLab
-Software modules: LAS X 2D Analysis/Multi Channel; LAS X 3D Visualization; LAS X 3D Analysis
