PoL – Physics of Life | Confocal & widefield microscopy

Zeiss LSM 980 with Airyscan 2

Zeiss LSM 980 is a point scanner confocal on a fully motorized Axio Observer 7 inverted microscope stand with a WSB scanning piezo stage (130 × 100 mm) and Z-Piezo 500 µm (1 nm resolution). The microscope is enclosed in a light-tight PECON enclosure (XLmulti S2 DARK) with temperature control (heating). It is also equipped with a stage top environmental chamber from PECON allowing temperature, CO2 and humidity control. Inserts for slides, dishes and multiwell plates are available.

Typical applications

  • Live-cell imaging and fixed samples
  • 2D and 3D cell culture
  • Zebrafish and fruit fly development
  • Long term timelapses
  • Cleared samples (RI: 1,33; RI: 1.47; RI: 1,5), using water or multi-immersion objectives.

Imaging modalities

  • Imaging of fixed sample with stitiching of large areas (tiling)
  • Widefield/Confocal/Super-resolution imaging
  • FRAP, Photo-activation
  • Spectral imaging (linear unmixing; FRET)
  • Fast imaging with Airyscan multiplex modes

Illumination

Lasers: 405/445/488/514/561/594/639/730 nm

Colibri 7 LED illumination (UV – 385/30 nm; V – 423/44 nm; B – 469/38 nm; C – 511/44 nm; G – 555/30 nm; Y – 590/27; R – 631/33 nm; FR – 735/40 nm).

Detection

Objectives

ObjectiveImmersionWorking Distance (mm)Cover Glass Thickness (mm)Best for
EC Plan-Neofluar 2.5x/0.085air8.80.17BF, Fluorescence (VIS, UV)
EC Plan-Neofluar 5x/0.16air18.50.17BF, DIC, Fluorescence (VIS, UV)
Plan-Apochromat 10x/0.45air20.17BF, DIC, Fluorescence (VIS, NIR-I)
EC Plan-Neofluar 10x/0.3air5.20.17BF, DIC, Fluorescence (VIS, UV)
Plan-Apochromat 20x/0.8air0.550.17BF, DIC, Fluorescence (VIS)
LD LCI Plan-Apochromat 25x/0.8multi-immersion0.570 - 0.17BF, DIC, Fluorescence (VIS, NIR-I)
C-Achroplan 32x/0.85water1.10 - 0.17BF, Fluorescence (VIS, NIR-I, 2-P)
Plan-Apochromat 40x/0.95air0.250.13 - 0.21BF, DIC, Fluorescence (VIS)
LD LCI Plan-Apochromat 40x/1.2multi-immersion0.410.15 - 0.19BF, DIC, Fluorescence (VIS, NIR-I)
C-Apochromat 40x/1.2water0.280.13 -0.20BF, DIC, Fluorescence (UV, VIS, NIR-I, 2-P)
LD C-Apochromat 40x/1.1water0.620.14 - 0.19BF, DIC, Fluorescence (UV, VIS, NIR-I, 2-P)
C - Plan-Apochromat 63x/1.4oil0.140.17BF, DIC, Fluorescence (UV, VIS, NIR-I, 2-P)
LD C-Apochromat 63x/1.15water0.600.14 - 0.19BF, DIC, Fluorescence (UV, VIS, NIR-I, 2-P)

 

Detectors

  • 2x Multi-Alkali PMTs
  • Spectral GaAsP PMT detector (32 channel)
  • Extended NIR GaAsP-PMT
  • T-PMT
  • Airyscan 2 (GaAsP)
  • Axiocam 712 mono camera (CMOS)

Add-ons

AI Sample Finder – automated sample and carrier identification, facilitating sample navigation and imaging.
Autoimmersion Module – automatic application of immersion media for water immersion objectives, streamlining the workflow, allows long term imaging with water objectives.
Definite focus 3 – focus stabilization and Z-drift compensation, allows for long term imaging.
FCS and FCCS – analysis of concentration and dynamics in the low molecular concentration regime.
Dynamics Profiler – analysis of concentration and dynamics, including flow directionality in the low–medium molecular concentration regime.
Spectral RICS – Image correlation based analysis of molecular dynamic at the cellular level​.

Software

ZEN 3.10 – Deconvolution, FRAP analysis, Experiment Designer, Tiles / Positions, Guided Acquisition, Direct Processing, Advanced Processing, Colocalization, Macro Environment.

www.zeiss.com/microscopy/en/products/light-microscopes/confocal-microscopes/lsm-980-with-airyscan-2.html

Gallery

Zeiss LSM 980 with Airyscan 2 – full view

Nikon Spinning Disk with CSU-W1 SoRa

The Nikon Spinning Disk is a microlens-enhanced Nipkow disk scanning confocal microscope on an inverted Nikon Ti2 stand, with an additional microlensed emission pinhole SoRa disk, a Mad City Labs Z-piezo (Nano-Z500) with 500 µm travel range and a motorized XY stage. The microscope is enclosed in an transparent OKO Lab H201-enclosure with temperature control (heating), and a stage top incubator with temperature, CO2, O2 and humidity control. This system is further equipped with a photomanipulation scanner module (FRAP, Photoswitching/-conversion) and a OMS - UV 355 nm pulsed 15uJ Laser Cutting and FRAP Unit.

Typical applications

  • Fixed and live samples, 2D and 3D
  • Fast acquisition
  • Long-term live imaging
  • Large field of view (600 x 600 µm2 for 25x magnification objective)

Imaging modalities

  • Widefield Epifluorescence
  • Spinning Disk Confocal
  • High Resolution in SoRa mode with up to 140 nm lateral resolving power (Super Resolution by Optical Pixel Reassignment.
  • Photomanipulation: FRAP, photoconversion, ablation, cutting

Illumination

Lasers: 405, 445, 488, 515, 561, 638 nm

CoolLED pE4000: 16 selectable LED wavelengths (365, 385, 405, 435, 460, 470, 490, 500, 525, 550, 580, 595, 635, 660, 740, 770 nm)

 

Detection

Objectives

ObjectiveImmersionWorking Distance (mm)Cover Glass Thickness (mm)Best for
CFI Plan Apochromat Lambda D 2X/0.1air8.50 - 0.17BF, Fluorescence (VIS, NIR-I)
CFI Plan Apochromat Lambda D 4X/0.2air200 - 0.17BF, Fluorescence (VIS, NIR-I)
Plan Apo λ 10x/0.45air40.17BF, DF, DIC, Fluorescence (VIS)
CFI Plan Apo VC 20x/0.75air10.17BF, DF, DIC, Fluorescence (VIS)
CFI Plan Apo λD 20x/0.8air0.80.17BF, DF, DIC, Fluorescence (VIS, NIR-I)
CFI Plan Fluor 20x/0.75multi-immersion0.51 - 0.330 - 0.17BF, DF, DIC, Fluorescence (UV, VIS, 2-P)
CFI Apo LWD λS 20x/0.95water0.99 - 0.90.11 - 0.23BF, DF, DIC, PhC, Pol, Fluorescence (VIS, NIR-I)
CFI Plan Apo λS 25x/1.05silicon0.550.11 - 0.23BF, DF, DIC, Pol, Fluorescence (VIS)
Plan Apo l 40x/0.95air0.21 - 0.160.11 - 0.23BF, DF, DIC, Fluorescence (VIS)
CFI Plan Fluor 40x/0.75air0.660.17BF, DF, Fluorescence (UV, VIS)
CFI Plan Apo λS 40x/1.25silicon0.30.13 - 0.21BF, DF, DIC, Pol, Fluorescence (VIS, NIR-I)
CFI Apo λS 40x/1.25water0.2 - 0.160.15 - 0.19BF, DIC, PhC, Pol, Fluorescence (UV, VIS)
CFI Apo LWD λS 40x/1.15water0.61 - 0.590.15 - 0.19BF, DF, DIC, PhC, Pol, Fluorescence (VIS, NIR-I)
CFI Apo TIRF 60x/1.49oil0.16–0.070.13–0.19BF, DIC, PhC, Pol, Fluorescence (VIS)
CFI SR Plan Apo IR 60x/1.27water0.18–0.160.15–0.19BF, DIC, PhC, Pol, Fluorescence (VIS, NIR-I, 2-P)
CFI Plan Apo VC 60x/1.2water0.31–0.280.15–0.18BF, PhC, Fluorescence (UV, VIS, 2-P)
CFI SR HP Plan Apo λS 100x/1.35silicon0.31–0.280.15–0.19BF, DIC, Pol, Fluorescence (VIS)

Detectors

  • Two Hamamatsu Fusion BT sCMOS cameras (high quantum efficiency and extremely low read-out noise)

Filters

DAPI, FITC, Cy3, Cy5, Cy7 Penta LED HC Set (Semrock)

 

Add-ons

  • Automatic Water Immersion Dispenser – automatic application of immersion media for water immersion objectives, streamlining the workflow, allows long term imaging with water objectives.
  • Enclosure with temperature control
  • Stage-top incubator – control of temperature, humidity, CO2 and O2 (including hypoxia conditions)
  • Perfect Focus System – focus stabilization and Z-drift compensation for long term imaging

 

Software

NIS 5.42 and NIS 6.10 – JOBS, ND Acquisition, Deconvolution, General Analysis 3, 3D Tracking, Calcium FRET, Illumination Sequence