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Center for Microscopy and Image Analysis

Widefield - Leica Thunder Imager 3D Live Cell (Irchel)

The LeicaTHUNDER Imager 3D Live Cell on the Leica DMi8 platform – a fully motorized, inverted research microscope equipped with hardware-based adaptive focus control (AFC) for precise drift correction and rapid focusing in multi-well applications. This versatile instrument is designed to capture fixed and live samples in up to 8 channels with remarkable speed. With THUNDER - Computational Clearing methodology, images are instantly deconvolved, offering confocal-like clarity while excelling in rapid time-lapse recording and minimizing phototoxicity. Built on the reliable DMi8 platform, it accommodates various sample types with stage holders for slides, petri dishes, and tissue culture plates. Imaging is acquired via a suite of high-NA objectives ranging from 5X to 100X. Two available cameras provide multi-channel fluorescence imaging via a high-sensitivity sCMOS B/W camera (Leica monochrome fluoroscence DFC9000 GTC) or high-resolution color imaging via a 12 bit CMOS RGB camera (Leica digital color camera DMC6200). Rapid switching between fluorescent dye channels is augmented by the use of a single quad-cube excitation filter set coupled with an external emission filter wheel. Experience enhanced live cell imaging capabilities, including precise focus maintenance over extended acquisition periods.

Responsible Person

responsible person

Location

University Zurich, Irchel Campus, Room Y42-H-93. 

Training Request

 Follow this link  to apply for an introduction to the microscope

The Thunder Imager features the following modalities:

Widefield

Widefield (WF) mode (sample illumination with LED)

DIC

Differential Interference Contrast (DIC) microscopy is another technique used to enhance contrast in transparent samples. It works by splitting a beam of light into two, altering their paths slightly, and then recombining them. This creates an interference pattern, making subtle variations in the sample's refractive index more visible. DIC microscopy produces images with three-dimensional appearance and enhanced contrast, making it useful for studying living cells and tissues in great detail.

Phase contrast

Phase contrast is a microscopy technique that makes transparent samples visible by enhancing the differences in how light waves pass through them. It helps to reveal details in objects like cells or tiny organisms that are hard to see with regular microscopy.

Fura-2

Fura-2 imaging is a method in cell biology  for tracking calcium levels inside cells using a special fluorescent dye, Fura-2. Fura-2 works by changing its glow under different lights based on how much calcium is present. When calcium binds to Fura-2, the dye's light emission changes. This allows scientists to measure calcium levels by shining light at two specific wavelengths (around 340 nm and 380 nm) and detecting the emitted light at a constant wavelength (about 510 nm). The difference in brightness between these two lights directly correlates with the amount of calcium, making it a reliable way to monitor calcium's role in cellular activities without being influenced by the dye's amount.

Technical Specification

Microscope body

  • inverted widefield microscope (Leica DMi8 stativ)

  • Quantum stage - high precision linear motor stage

    • travel range 120 x 80 mm

    • position resolution 5 nm

    • max speed 500 mm/s

    • max. acceleration 2 m/s2

Light Sources and Lasers
  • Leica LED8
    • 390 nm
    • 440 nm
    • 475 nm
    • 510 nm
    • 575 nm
    • 635 nm
    • 747 nm
  • CoolLED pE340fura for Calcium Imaging
    • 340 nm
    • 380 nm
  • Transmitted light LED light source

Camera System
  • Leica monochrome fluoroscence DFC9000 GTC sCMOS
    • Pixel: 2048 x 2048 (4.2 Megapixel)
    • Pixel size: 6.5 µm
    • 19 mm sensor diagonal
    •  82% quantum efficiency max. at 580nm
    • Sensor chip is air cooled to 0°C
    • 90 fps acquisition speed
    • Bit depth: 12/16 bit
    • Hardware binning: 2x2, 3 x3,4 x 4, 8 x 8
  • Leica digital color camera DMC6200
    • 2.3 Mpixel resolution up to 20.7 Mpixel with pixel shift
    • CMOS Sensor with 5.6 µm pixelsize
    • up to 40 fps with 2.3 Mpixel; 3x16 bit color depth

Environmental control

  • Incubator i8 with heating unit (heats up to max. 30° C above ambient temperature)

  • CO2-O2 controller:

    • Range of CO2 Concentration: 0 - 20 Vol%

    • Range of CO2 Concentration: 0 - 21 Vol% Resolution of CO2 and O2 displays: 0.1 Vol% Resolution of internal

    • CO2 and O2 loop control: 0.01 Vol%

    • Actual minimal O2 concentration at the sample 0.8 % 

  • Incubator XL dark and top stage incubation (full environmental chamber, temperature and CO2 control)

Accessories
  • PECON KM Frame for well plates
  • PECON Click-In for petri dishes (35 mm and 60 mm)
  • PECON Click-In for standard object slides, µ-Slides by Ibidi, Nunc Lab-Tek Slides and Chambers by Thermo Scientific
  • Matrix screening software for more complex experiments
  • Ludin chambers for 12 mm or 18 mm coverslips

Available Optics
Name Magnification NA Immersion WD (mm) Pixel size
LEICA N Plan PH0
5x
0.12
Air
814
1.2939 x 1.2939 µm
LEICA  HC PL Fluotar PH1
10x
0.3
Air
11
645 x 645 nm
 xLEICA PL FL L CORR PH1
20x
0.4
Air
6.9
320 x 320 nm
LEICA HC PL APO CORR CS2 (on request available)
20x
0.75
Air
0.67
-
LEICA HC PL APO CORR
40x
0.95
Air
0.17
161 x 161 nm
LEICA PL Fluotar CORR PH3 (on request available)
63x
1.1
Oil
0.35
-
LEICA  HC PL APO CS2 
63x
1.4
Oil
0.14
102 x 102 nm
LEICA  HC CORR CS (on request available)
100x
1.44
Oil
-
-
LEICA  PL APO PH3 
100x
1.4
Oil
0.09
64 x 64 nm

 

Available Filters

  
Name Excitation filter Main beam splitter Emission Filter
DFT5 Quad filter
  • 375-407
  • 462-496
  • 542-566
  • 622-654
  • 415
  • 500
  • 572
  • 660
  • 420-450
  • 506-532
  • 581-607
  • 666-724
CYR7  Quad filter
  • 422-450
  • 495-517
  • 566-590
  • 710-750
  • 459
  • 523
  • 598
  • 763
  • 462-484
  • 527-551
  • 602-680
  • 770-850
FURA2
  • BP 340/26, 380/11
  • 400
  • BP 510/80

Emission clean-up filterwheel (external):
 

Position Range

1

 460/80
2 535/70
3 590/50
4 642/80
5 empty=100% transmission

 

THUNDER

THUNDER Imagers feature the innovative Leica technology Computational Clearing. It efficiently removes out-of-focus blur in real time, enabling the meaningful use of 3D specimens with camera-based fluorescence microscopes. The high sensitivity of the system ensures low phototoxicity and photobleaching, i.e., higher throughput with optimal conditions.

Instant Computational Clearing (ICC) Instantaneous removal of out of focus background
Small Volume Computational Clearing (SVCC) Addition of 3D deconvolved decision mask on ICC, best for thin specimens
Large Volume Computational Clearing (LVCC) Addition of 3D deconvolved decision mask on ICC, best for thick specimens

 

User Guide

ZMB-Guides - Wild-field LEICA THUNDER

Links & Literature

Leica Thunder Imager 3D Cell culture
 

Computational Clearing

Bereichs-Navigation

Unterseiten von Widefield - Leica THUNDER (Irchel)