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OptionsNL5: The Latest in Line-Scanning Confocal Microscopy
NL5 is a new line-scanning confocal microscopy system offering an unprecedented combination of speed, sensitivity & resolution. It allows you, for example, to quickly screen a multi-well plate with multicolor images, to select the most promising ones. Because its design is based on line scanning, the NL5 system combines the scanning speed of spinning disc techniques and the resolution of laser point scanning.
Capture 3D live samples with minimal phototoxicity, high resolution and contrast. Like RCM, NL5 is easy to operate and keeps your samples happy under the microscope for a long time. Convert your research microscope into a fast-scanning confocal microscope that provides very gentle conditions for your live samples.
Contrary to spinning-disk systems, another advantage of NL5 and its slit pinhole is that it does not lead to pinhole crosstalk when imaging deep within the sample. NL5 reaches a lateral resolution of 240 nm, which can be easily converted into 170 nm with on the fly deconvolution.
NL5 is an excellent add-on to any research microscope. It allows for converting a wide-field system into a fast scanning confocal with high resolution and sensitivity.
Fast Scanning Speed, Low Levels of Phototoxicity
Some natural processes in the field of biology happen fast, like cell and protein dynamics and cell-cell interactions. Therefore, fast scanning speed is needed to study them, which is exactly what NL5 provides. In addition, researchers in developmental biology, regeneration or immunology fields often need to image large live samples (mouse embryos, zebrafish, organoids) for a long time without losing any detail of the cell’s behavior. This is why NL5’s combination of speed and low phototoxicity is the perfect solution.
The Benefits of Using NL5
NL5 is a fast scanning confocal system that reaches 55 fps (full frame), surpassing the speed of conventional point-scanning confocal systems. In addition to speed, it brings very gentle imaging conditions for your samples. Perform live cell imaging for more than a day, with no signs of phototoxicity.
NL5 does not cause any pinhole cross-talk, as it has only one slit pinhole. Pinhole cross-talk is associated with spinning disk systems, because they contain a disk with multiple pinholes combined and the out-of-focus light can sometimes pass through the closeby pinholes. This effect is more evident when imaging deep within the sample, making NL5 the ideal system for those applications.
NL5 is a cost-effective solution that can be coupled to any research microscope via the c-mount interface
Who Should Use NL5 and Why?
NL5 is a compact, easy-to-use line-scanning confocal microscopy system, making it a great addition to any microscopy facility. It’s a fruitful solution for individual research labs to easily convert any existing widefield fluorescence microscope into a fast confocal microscope system with high sensitivity and resolution. Capture datasets of 170nm lateral resolution after deconvolution at 25 fps. Study fast cell and molecular dynamics, or perform high contrast deep 3D imaging with minimal aberrations. The high sensitivity of the modern sCMOS cameras used as detectors facilitates acquisition with low light sample exposure, making NL5 ideal for live cell imaging
Convert your existing research microscope into a fast-scanning confocal system ideal for deep 3D live cell imaging.
- Single digital scanner (same as RCM2)
- 170 nm resolution in real time (microvolution)
- Extreme sensitivity
- Low phototoxicity
- NL5 runs at a speed of 25 fps
- High contrast deep 3D imaging without pinhole cross-talk
- High sensitivity and resolution (170 nm after deconvolution)
- Live cell friendly
- Hardware integration into third-party software
- Compact & easy to use
To discover our whole line of confocal microscopy add- ons, click here .
*Videos above:
(1) Max projection of a stack in time with 1000 frames per color HUVEC labeled for SPY505-DNA, SPY555-Actin and SPY650-Tubulin Sample courtesy of Philippa Phelp (Boon’s Lab) Physiology department. VUmc Amsterdam
(2) Zebrafish embryo (24 to 48 hpf). Endothelial cells labelled with GFP Sample courtesy: Kinki Jim. VUmc Amsterdam, MMIC
(3) HeLa Cell with cytosolic GFP. Sample courtesy of Caroline Pham (Lodder’s Group). Experimental Cardiology Department. VUmc location AMC, Amsterdam
NL5 Applications
Fast Cell Dynamics
- Better visualization and analysis of thin and dynamic structures even with dim signals
- Study fast cell dynamics with very low phototoxicity
- Cell migration
- Intra/extra-cellular transport
- Cell signaling
Live Cell Imaging
- Imaging happy cells and organisms for more than a day
- Long-term live cell imaging in gentle conditions
- Low phototoxicity and bleaching
- Developmental biology
- Regeneration
- Immunology
Deep 3D Imaging
- One slit pinhole for better images in deeper frames
- Organoids
- Zebrafish/mouse embryos
- 3D culture with hydrogels
White Papers:
Acquisition Software:
Both NL5 and NL5+ are compatible with Micro-Manager, InScoper, Volocity & NIS-Elements software for acquisition.
Deconvolution Software:
Both NL5 and NL5+ are compatible with Microvolution and SVI Huygens software for deconvolution.
Compatible Devices
NL5 and NL5+ are confocal add-ons. To get a fully functional system a microscope stand, a camera and a laser launch are required. Below is a non-exhaustive list of devices compatible with NL5 and NL5+.
Microscope Stands:
- Leica DMi8
- Nikon TE2000
- Nikon Ti-U & Ti-E
- Nikon Ti2-U & Ti2-E
- Olympus IX71 & IX81
- Olympus IX73 & IX83
- Zeiss Axio Observer.Z1
Cameras:
- Hamamatsu ORCA-Flash4.0 LT+, LT3 & V3
- Hamamatsu ORCA-Fusion & Fusion BT
- Hamamatsu ORCA-Fire
- Hamamatsu ORCA-Quest
Laser Combiners:
- Oxxius L4Cc & L6Cc
