Zeiss 800 Alryscan Confocal Microscope

Zeiss 800 Alryscan Confocal Microscope

 

The LSM 800 microscope is equipped for a wide range of applications and techniques, including:

  1. Airyscan detector for improved resolution in 3D
  2. Emmision fingerprinting
  3. Time lapse imaging of live cells (stage-top incubation)
  4. High content screening.
  5. Differential interference contrast (DIC) in combination with fluorescence
  6. Polarized light microscopy
  7. A range of different objectives from 2.5x to 63x magnification

 

The microscope

Axio Observer.Z1 inverted microscope with Definite Focus for adaptive focus control. Automated microscope with a motorized XY-stage and touch pad.

 

Focus drives

  • Motorized objective nosepiece (z-step size 10 nm)

 

Scanners

  • Galvo scanning, up to 8 frames /sec at 512 x 512 pixels image format can be achieved.

 

Objectives

 

Magnification NA Type Immersion Coverslip WD (mm)
2.5x 0.085 EC Plan Neofluar Air 0.17 8.8
10x 0.3 EC Plan Neofluar Air 0.17 5.6
10x 0.45 C-Apochromat Water 0.17 1.8
20x 0.5 Plan Neofluar Air 0.17 2.0
20x 0.8 Plan Apochromat Air 0.17 0.55
25x 0.8 LCI Plan Neofluar Imm/Corrr -/ 0.17 0.21
40x 0.75 Plan Neofluar Air 0.17 0.71
40x 0.8 C-Achroplan NIR Water 0.17 1.70
40x 1.2 C-Apochromat Water 0.14 - 0.19 (Corr ring) 0.28
63x 1.15 LD C-Apochromat Water 0.14 - 0.19 (Corr ring) 0.60
63x 1.2 C-Apochromat Water 0.14 - 0.19 (Corr ring) 0.28
63x 1.4 Plan Apochromat Oil 0.17 0.19
  • The 25 x 0.8 objective is a glycerine, silicone and water immersion objective
  • DIC prisms exists for the following objectives: 20x/0.5, 40x/075 and 40x/1.2 W

 

Condenser lens

NA 0.55, air, working distance 26 mm

 

Lasers

  • LSM 800 Laser Module URGB, diode lasers 405 nm (5 mW), 488 nm (10 mW), 561 nm (10 mW) and 640 nm (5 mW)
  • An acousto-optical tunable filter (AOTF) controls the intensity of the visible excitation laser light
  • One main beamsplitter is used for all laser lines to separate excitation light from the emitted photons

 

Detectors

Three internal GaAsP detectors (with pinhole), PMT 1- PMT 3, in reflection mode with tunable emission bands in the range 400 – 800 nm. PMT 3 is the Airyscan detector, a hexagonal GaAsP detection array.

One external PMT (without pinhole) in transmission mode.

 

Light for epi-fluorescence and transmitted light

  • External HXP 120W, fiber optics and bulb lifetime 2000h
  • Halogen lamp: 12V – 100W

 

Filter cubes for epi-fluorescence

 

Name Excitation filter Emission filter Dichroic
UV filter set/ DAPI BP 340 - 400 nm LP 420 nm 400 nm
Filter Set 09/ AF 588 BP 450 - 490 nm LP 515 nm 510 nm
Filter Set 15/ AF 543 BP 540 - 552 nm LP 590 nm 580 nm

 

Camera

Zeiss Axiocam 503 color camera, 2.8 Mpix, 38 fps (color).

 

Software on the LSM 800 computer

Zeiss ZEN Blue with software modules

  • Tiles/Positions/Regions
  • Airyscan
  • Time Series
  • Z-stack

 

Accessories

  • Stage-top incubation system from Pecon (heating inserts for petri dishes diameter ~50 mm and Ibidi/LabTek chambers. Warm air incubation and CO2-control).
  • Hardware-based focus stabilizer