The newest addition to the Imaging Core is the Leica THUNDER Live Cell Fluorescent Compound Microscope. Installed in December of 2021, this instrument is designed to image fixed and live samples in up to five channels with unprecedented speed and clarity. Images are recorded with one of two high sensitivity and high-resolution widefield cameras, and then instantly deconvolved utilizing Leica’s proprietary Computational Clearing methodology to generate near-confocal clarity. The speed of widefield imaging outperforms scanning confocal imaging in areas of rapid time-lapse recording, Calcium imaging, and reduction of phototoxicity.
The THUNDER Live Cell microscope is built upon Leica’s workhorse DMi8 fully-motorized inverted microscope, with stage holders for microscope slides, large and small petri dishes, and tissue culture plates. Imaging is acquired via a suite of high-NA objectives ranging from 5X to 40X, all of which are dry to allow for rapid switching from low to high magnification without having to disturb the sample. Two available cameras provide multi-channel fluorescence imaging via a high-sensitivity sCMOS B/W camera or high-resolution color imaging via a 12 bit CMOS RGB camera. 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. Light phototoxicity is reduced with an advanced real-time controlled 7-line LUMENCORE LED light source. All components of the system are further speed-optimized by the use of an advanced real-time controller that synchronizes the stage movement, illumination, focus controls, and camera shutters. Live cell imaging is enhanced with a suite of tools that keep the samples in focus over long periods of acquisition, and even track cells as they move over time.
Leica DMi8 fully motorized inverted microscope with hardware Automatic Focus Control for drift compensation and closed loop focus system with Z reproducibility of better than 20 nm. Quantum Stage for high speed and robust position repeatability. Stage inserts for standard microscope slides, 30 and 90 mm petri dishes, and tissue culture plates. Motorized polarizer and DIC turrets for polarized, phase contrast, and Differential Interference Contrast optics. Advanced real-time controller for high speed synchronization of stage, illumination, focus and camera components. Leica motorized SmartMove stage controller for remote stage movement.
Objective | Magnification | N/A |
N Plan 5x Ph 0 | 5 X | 0.12 |
HC PL Fluotar 10x Ph 1 | 10 X | 0.32 |
HC PL Fluotar L 20x Corr Ph 1 | 20 X | 0.40 |
HC PL Apo 20x | 20 X | 0.80 |
HC PL Fluotar L 40x Corr Ph 2 | 40X | 0.60 |
PL Apo 40x Corr | 40X | 0.95 |
Light Source | Output lines |
Lumencor Spectra X LED | Tunable 380-680 nm output for DAPI, GFP, YFP, TRITC/mCherry, and Cy5. 7 user defined bandpass filters |
Filter | Dyes |
Quad Cube | DAPI, FITC, TRITC, Cy5 |
DAPI | DAPI, Hoechst |
FITC | FITC, GFP |
TXR | Texas Red, mCherry, RFP, DSRed |
Camera | Specifications |
Leica DFC9000 GTC | 4.2 MP sCMOS, 6.5 micron pixel size, 19 mm sensor diagonal, 82% quantum efficiency, dark current to 0.14e-/p/sec, 90 fps acquisition speed, extended dynamic range of 33000:1 |
Leica K3C | CMOS, 2.4 micron pixel size, 8.92 mm sensor diagonal, 15 fps acquisition speed, dynamic range up to 72 dB, 3X12 bit RGB, 12 bit mono. |
Module | Uses |
Navigator | GUI for point scanning and Tiled images |
Assay Editor | Automatic distribution of ROIs and positions within defined sample carrier for statistical acquisition |
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 |
3D Visualization Advanced Module | 3D visualization and rendering of Z stack files. Display clipped and orthogonal sections, stereo projections, and 3D movies. |
3D Analysis | Wizard for processing and measurement of 2 and 3 dimensional images, generate reusable workflows for processing and analysis of multiple data sets. |
Microlab | Wizard for Z stacks, timelapse |
Intel XEONW-2123 3.6 4C CPU with Win10 64-Bit
64GB DDR4 RAM and High end graphic card
2TB SSD-RAID
SSD1 (512GB)
Applications:
Fluorescent multi-channel imaging; High Speed imaging, Brightfield, DIC and Phase Contrast color imaging, Live cell imaging with timelapse/Automatic focus/cell tracking; tilescanning, point scanning, multiwell plate scanning; adaptive deconvolution.
Output:
.lif, .tiff, .jpg, .avi, .mov, .mpeg4