Leica Thunder Wide Field Fluorescence Microscope with Computational Clearing and Deconvolution

Microscope

Some info:  THUNDER Revealing Image Detail Hidden in the Haze , Overview PDF , Spheroid-Imaging The system is built on a fully automated, inverted Leica DMI8 microscope equipped with AFC (hardware autofocus control) and the objectives below.

Objectives:

  • N PLAN 5 x / 0.12  Dry
  • HC PL Fluotar 10 x / 0.32 Dry
  • HC PL Fluotar 20 x / 0.40 Dry
  • HC PL Fluotar 40 x / 0.60 Dry
  • HC PL APO 63 x / 1.40 Oil

Fluorescence Filters

Name LED Excitation filter Dichroic mirror Emission filter
CFP 440 BP 436/10 455 BP 480/40

LED Excitation, Quad Filter Block (Dichroics/Filters) and Filter Wheel

Two multiband filter cubes are available (see below). The first is a quad filter set that uses an external filter-wheel to avoid crosstalk between different dyes. The second is a standard multiband filter cube for cyan, yellow, red and near infrared emission dyes).
Quad Filter Block LED Excitation Range Dichroic Emission Range Emission Filter wheel FLUOROPHORES
DFT51010 390 375-407 415 420-450 BP440/40 DAPI, Hoechst
475 462-496 490 506-532 BP510/40 FITC, GFP, Alexa488
555 542-566 572 578-610 BP590/50 Alexa568, Rhodamine, DsRed-2
635 622-654 660 666-724 BP705/72 Alexa647, Cy5
CYR70010 440 422-450 459 462-484 100 % CyanFP
510 495-517 523 527-551 100 % YellowFP
575 566-590 598 602-680 100 % mCherry, Alexa594, Texas-Red
747 710-750 763 770-850 100 % nIR Autofluorescence

Motorized XY Stage ITK QUANTUM

For fast and precise sample positioning and mosaic/multiposition acquisitions (see specifications below). It is provided with several inserts to hold slides, plates and 96-multiwell plates.
  • Travel range 122 x 82mmSpeed 500mm/sec
  • Resolution 5 nm
  • Reproducibility < 1 μm
  • Accuracy < +/- 1 μm

Leica DFC9000 GTC sCMOS camera

  • 4.2 MPixel
  • Image sensor type: Front-illuminated scientific CMOS CIS2020A
  • Shutter: Rolling shutter with global reset
  • Pixel: chip of 2048x2048 pixels; 6.5 mm pixel size
  • Speed of Acquisition: 50 fps (USB 3.0); 90 fps (Camera Link)
  • Maximum quantum efficiency: 82% @580 nm
  • Binning: 2x2, 3x3, 4x4, 8x8
  • 12 bit and 16 bit digitalizers

Environmental control

UNO Stage-Top Incubator (OKO-LAB). Includes #H301-K stage incubator equipped with inserts for 2 petri dishes and 1 slide, universal multi-well plates, 12-well plates (and more). CO2 and humidity are regulated through the manual UNO-H-T-CO2 controller (OKO-LAB) equipped with an Air/CO2mixer, an air pump and a humidity module.

Thunder Algorithms

3 dedicated algorithms are available that remove out-of-focus light in real-time, thus providing fast optical sectioning capabilities:
  • Instant Computational Clearing (ICC): Removal of background and out-of-focus light.
  • Small Volume Computational Clearing (SVCC): ICC and deconvolution algorithm for thin samples (e.g. cells, yeast)
  • Large Volume Computational clearing (LVCC): ICC and deconvolution algorithm for thick samples (e.g. tissue sections, embryos, organoids).
  The software used is Leica LAS-X