Super Resolution Imaging Solution Provider. Imaging Future

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1 Super Resolution Imaging Solution Provider Imaging Future

2 Imaging Solution More Than Equipment NanoBioImaging(NBI) is the Industrial Partner of HKUST Super Resolution Imaging Center (SRIC). NBI aims to develop and commercialize unconventional optical microscope techniques and provides professional, affordable and fast super resolution imaging services for life sciences institutes, universities and the biomedical industry.

3 Super Resolution Imaging - A Door To The Nano-World Using appropriate dyes, the fluorescent signal decay can be combined with single molecule localization techniques to achieve 20 nm resolution. The NBI super resolution (SR) imaging system achieves a 10-fold improvement compared to conventional confocal microscopy. It allows nanoscale quantification of protein localization or protein-protein interaction. The super resolution microscope can be used for biological research in cell biology, neuron biology, immunology, virology and nanomaterials. Golgi Apparatus a b Two-color super resolution imaging resolves GM130 (cis-golgi matrix protein) and Golgin97 (trans-golgi network membrane protein) in HeLa cell line. (a) Epi fluorescent image with blurred strucutre and more overlapping regions of the two proteins. (b) Super resolution image at the same location where two layers (cis vs. trans) in golgi apparatus are better distinguished. (Collaboration with biochemistry group from HKUST) Yeast Organelle Widefield Image Super Resolution Image F-actin Network Structure Two-color super resolution imaging of Alexa750-anti-GFP nanobody labeled Tub1 of spindle microtubule and Alexa647-anti-RFP nanobody labeled Tub4 of spindle pole body in a yeast cell. (a) and (b) are Epi fluorescent images where (b) is the zoom in of the marked area in (a). (c) and (d) are the two-color super resolution images at the same position where the anchoring of spindle microtubules to spindle pole bodies can be clearly resolved. (Collaboration with biochemistry group from HKUST) Three-dimensional super-resolution imaging of F-actin network in Drosophila S2 cell, with the z-position information color-coded according to the color scale bar. (Collaboration with biochemistry group from HKUST) 1000 nm Neuron Synapse 1 μm 1 μm Synaptic structure in tissue slice resolved by localization microscope. (a)(c) and (e) show the EPI-fluorescent image which fails to show structural details. (b)(d) and (f) are super resolution images acquired by NBI two-color localization microscope. The pre- and post-synaptic structures are clearly resolved. (Collaboration with biochemistry group from HKUST) Mitochondrial Membrane Structure Two-color super resolution imaging resolves p32 (mitochondrial matrix) packed by Tom20 (mitochondrial outer membrane) in HEK293 cell line. (a) Epi fluorescent image with totally blurred structure. (b) The super resolution images at the same location. (c) The transverse profile correspondence to boxed region in (b), clearly shows that the p32 is packed inside the mitochondrial outer membrane. 200 nm 200 nm 200 nm 200 nm (Collaboration with biochemistry group from HKUST)

4 How to Obtain The Best SR Imaging Result? Super resolution imaging is a combination of physics, chemistry, biology, computer science and engineering technology. This interdisciplinary nature can make it challenging for biologists (experimental design, sample preparation, equipment calibration, software configuration, data analysis). In collaboration with HKUST SRIC, NBI has built a team that provides customized hardware, software and comprehensive technical support. Ultimately our customers benefit when they get the best results with minimal effort. Stable and Efficient Hardware Local and Instant Tech-support Customized & Upgradable System Design Super Resolution Imaging Plan Exclusively For You Professional & Comprehensive Service Support Easy and Convenient Software Professional and Accurate Analysis NBI-SR Widefield

5 Stable and Efficient Hardware Architecture of the super resolution imaging system developed by NBI: STABLE High-Resolution Sample Locking Real-time locking system compensates lateral sample drift in caused by environmental conditions (temperature change, air flow, vibratior noises). Compared with post processing techniques used by others, the sample locking module designed by NBI can compensate this drift at nanometer resolution with high reliability. System Specifications EFFICIENT Two-Channel Imaging Simultaneously The intermediate image is split and imaged on EMCCD sensor through two separate optical paths simultaneously. Compared with general super resolution microscopy, long term acquisition of super resolution imaging benefits from the two channel design, which reduces photobleach and halves the imaging time. Super-Resolution Compatible Laser Color Channels Compatible Objective Compatible Microscope Camera Autofocus Lateral Locking Anti-vibration Environment Dye ~ 20 nm in lateral ~ 50 nm in axial 405 nm, 488 nm, 656 nm, 750 nm and others upon demand TTwo channel imaging simultaneously CFI Apochromat TIRF 100 oil (NA1.49) CFI Plan Apochromat VC 100 oil (NA1.40) ECLIPSE Ti-E Andor ixon 897 EMCCD Active Pixels Pixel Size 16 microns Readout Noise < 1e- Perfect Focus System LED based Active Locking, std ~ 1nm Optical table with passive isolator General lab conditions Alexa 647, Alexa 750 and others upon demand

6 Convenient and User-Friendly Software ROHDEA is software developed by NBI for super resolution imaging. It makes using the system both easy and efficient for our users. Automatic User & Data Management Easy to Test Arguments for Reconstruction The parameters for the reconstruction algorithms will affect the result quality. The testing step offers user an easy and visualized way to test different arguments for better configuration. Streamlined Image Acquisition ROHDEA maintains all data for each user in an intelligent and wellorganized structure, including raw images and all useful parameters and computed/reconstructed results. ROHDEA also records configuration of arguments for similar experiments in the future. ROHDEA provides improved user experience compared with competing imaging software solutions that are function-oriented and are harder to use. ROHDEA structures super resolution imaging workflow into four steps: 1. view preparing, 2. wide-field image acquisition, 3. super resolution image testing and 4. super resolution image acquisition. This standardized sequence of steps is easier to follow compared with traditional approaches. Simplicity is the soul of efficiency Real-time Monitoring of Reconstruction with Details Imaging Result Analysis Users can analyze the reconstructed super resolution images with SPRUCE, an image analysis tool also developed by NBI, to get customized analysis reports. Analysis of the result can provide useful information such as the distribution of cluster sizes/ patterns, localization distribution, error distribution etc. ROHDEA embeds multiple super resolution imaging algorithms, and presents real-time reconstructed images. Parameters can be changed in real-time to optimize the result.

7 Professional and Comprehensive Services The ultimate goal of microscopy is to achieve high quality images. NBI imaging service will take you there with minimal time and effort. Experimental Design - Super Resolution Or Not Judging whether a new technology will be useful for a particular application requires experience. NBI can give you the answer rapidly, with our academic strengths and our knowledge base in the field. We also customize an imaging plan for your project. You can also choose our imaging service, with which you can test your hypothesis within weeks. Contact us now to discuss your imaging project! Training - More Than Operation Long Term Collaboration - Make Progress Together Leading edge technologies can provide an advantage in research. In our long-term technical collaboration, our customers have priority access to our most advanced technologies. Furthermore, NBI also provides customized solutions for your research needs. NanoBioImaging Help Know Why, Not only How Following a standard "step-by-step" manual process can produce the optimal result for a demo sample. However your particular samples may require a different approach. NBI's scientists in different fields can provide professional training to the users, so they know which technique will work best for their situation. Consultation - Your Problem Is Our Problem Hardware after-sales service doesn't just mean hardware warranty -- helping you run your machine is also considered to be an important part of NBI service. Our consultation service includes trouble shooting, sharing technical experience, data analysis and modeling, statistical analysis, etc.

8 Imaging Future Unit 623, 6/F Biotech Centre 2, No.11 Science Park West Avenue Hong Kong Science Park, Shatin, N.T., Hong Kong Tel: (852) (86) Specifications and appearances are subject to change without notice or obligation on the part of the manufacturer.

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