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DeepVision™ Imaging Instrument for Novel NIR-I and NIR-II Dyes and Probes
An introduction video on DeepVision™ Imaging System and its technology
(Also see dual modality KingsVisionTM with bioluminescence imaging capability. Also see Nirmidas NIR-I and NIR-II dyes and probes.)
In vivo fluorescence imaging of small animals has been done typically in the NIR-I (800-900 nm) range, suffering from shallow imaging depth and high background due to light scattering and tissue autofluorescence. NIR-II/SWIR imaging is a breakthrough development detecting fluorescence/luminescence in the 1000-1700 nm range to suppress these effects, affording single cell resolution at down to ~ 3 mm depth and useful feature resolution up to ~ 1 cm depth. Combined with ultra-bright NIR-II probes (molecules, quantum dots and rare-earth nanoparticles licensed from Stanford University) from Nirmidas Biotech and a new camera technology (10X shorter exposure time and lower noise than older brands/makers), DeepVision™ affords high performance non-invasive imaging of vasculatures, tumors, intact mouse brain, lymphatic vessels/lymph nodes and molecular imaging using antibody conjugated probes. It is a new generation of NIR imaging instrument empowering researchers to interrogate cardiovascular, cancer, brain and immune disease models. NIR-II/SWIR imaging is generating tens of thousands of publications in recent years and is ideal for deep-tissue animal imaging in vivo. |
In addition to CW imaging mode, a unique added feature of DeepVision™ is its fluorescence lifetime imaging mode. By setting a delay time of detection, the emission generated by a 975 nm excitation pulse from rare-earth nanoparticles (lifetime ~ 10 milliseconds) versus the emission from quantum dots (lifetime in micro-second range) can be distinguished despite their overlapping emission wavelengths. Such life-time imaging detects zero background. This scheme can also be used for multiplexed molecular imaging in the least scattering 1500-1700 nm NIR-II window for single cell biomarker profiling in vivo.
Key Features
A high performance and cost-effective biological imaging system detecting in the NIR-II/SWIR (1000-1700 nm) optical window. It is also capable of imaging fluorescence in the visible (600-700 nm) and NIR-I (800-1000 nm) windows. KingsVision™ also provides bioluminescence imaging, upconversion imaging and comes with two cameras.
Multiple and customized lasers (808 nm and 975 nm as default)
For deep tissue, high signal/background ratio in vivo small animal imaging, ex vivo and in vitro organ, tissue and cell culture imaging
Equipped with low magnification and high magnification microscopy imaging modes for whole mouse-body and single cell/single vessel imaging with switchable lens sets
Up to 120 frames per second video rate imaging
Ultra-low noise camera
Lifetime imaging capability
Multi-color, multiplexed molecular imaging using organic and nanoparticles probes emitting up to 1700 nm
Provides a unique demo sample containing micro-array printed dots of fluorescent probes emitting in the > 1100 nm and > 1500 nm respectively for rapid calibration of the system and for training.
Inside the DeepVision™
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Imaging Chamber
Light-tight imaging chamber
808 nm and 975 nm lasers by default (customizable).
Emission filter wheels – 3 default filters 1100 nm, 1300 nm and 1500 nm long pass.
Heating pad for mouse; temperature can be adjustd to keep mouse warm
Adjustable imaging field for whole body or high-resolution microscopic imaging in vivo. Three optical paths for 1X, 2.5X imaging and for microscope mode imaging at up to 50X magnification for single vessel and single-cell resolution in vivo.
Mouse stage can hold 4 mice at a time.
X-Y-Z control for mouse stage by joy stick. X-Y motion can switch from mouse to mouse and move from site to site of the same mouse. Z motion controls focus.
Video rate imaging capability with up to 120 frames-per-second.
Instrument integrates a white light for open-door surgery or manipulation.
DeepVision™ Imaging System Specification
Imaging System Components | Specifications |
Camera1 Active Area | < 1 cm x 0.8 cm |
Imaging Pixels | 640 x 512 |
Quantum Efficiency | 80%-90% in NIR-II 1.0-1.7 µm; 50-80% in 0.8-1.0 µm |
Pixel Size | 15 µm x 15 µm |
Minimum Field of View (FOV) | 1.0 mm x 0.75 mm (microscopy mode) |
Maximum Field of View (FOV) | 62 mm x 50 mm (whole body mode, up to four mic) |
Highest Spatial Resolution | < 2.5 µm |
Stage Temperature | 20 °C – 40 °C, heated stage. 37°C by default. |
Power Requirements | 110 or 220 V, 50 Hz – 60 Hz |
Dark Current | < 1500 e-/p/sec |
Read Noise | < 20e- (High Gain) <180e- (Low Gain) |
Lasers2 | 808 nm and 975 nm (for NIR-II and upconversion imaging) |
Emission Filters3 | 1100 nm long pass, 1300 nm long pass and 1500 nm long pass |
CCD Operating Temperature | -15 °C to -30 °C |
Imaging System Space Requirements | 61*60*96 cm (W x D x H) |
Imaging Modes | NIR-I imaging, NIR-II imaging, Bioluminescence imaging, Upconversion imaging, Lifetime imaging |
Lifetime Imaging Mode | User specified laser excitation time, laser off, followed by specified exposure/imaging time |
1. Nirmidas uses a new camera technology with superior performance, allowing for much shorter exposure time and lower background.
2. Different laser lines are available upon request
3. Different filters are available upon request
Application Examples
Whole Body Imaging
Fig2. A mouse inoculated with a 4T1 tumor was sequentially injected of carbon nanotube (CNTs) and an organic probe p-FE. Utilizing the fluorescence of p-FE emitting in 1100–1300 nm to highlight vasculatures (green) and that of laser-ablation produced CNTs emitting in 1500–1700 nm to highlight the tumor (red), mouse tumor model in vivo can be profiled.
Data from Wan et al., Nature Comm, 2018
Through scalp/skull non-invasive video rate imaging of cerebral vessels in a mouse head.
The fluorescence signal of rare earth nanoparticles in the cerebral vessels was observed through the scalp and skull of mice. During the video recording, the mouse scalp was slightly moved, and the transparency and three-dimensional structure of the mouse head were clearly visible.
In Vivo Two-Plex Molecular Imaging at ~ 1600 nm based on Fluorescence Life Time
Fig 3. Two-plex molecular imaging of immune response.
Top panel:
The principle of distinguishing short-lived PbS QD fluorescence from long-lived ErNP rare-earth nanoparticle luminescence.
Lower panel:
Two-plex imaging of a CT-26 tumor on a mouse with intravenously injected ErNPs-antiPDL1 (green) and PbS-antiCD8 (red). The zoomed-in high-magnification outlines the tumor with micrometer image resolution.
Modified from Zhong et al., Nature biotechnology, 2019
References
Molecular Imaging of Biological Systems With a Clickable Dye in the Broad 800- To 1,700-nm Near-Infrared Window. PNAS, 114(5):962-967, 2017.
A Bright Organic NIR-II Nanofluorophore for Three-Dimensional Imaging Into Biological Tissues. Nat Commun, 9(1):1171, 2018.
In Vivo Molecular Imaging for Immunotherapy Using Ultra-Bright near-infrared-IIb Rare-Earth Nanoparticles. Nat Biotechnol, 37(11):1322-1331, 2019.
Nirmidas DeepVision™ is available to purchase today. Email us at info@nirmidas.com or fill in the form below.
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