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 |
| 730 |
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
