The spatial resolution of conventional microscope is about 250nm in lateral direction, and 800nm in axial direction. NBI applies localization microscope technology in SRiS system, which can achieve 20nm in lateral direction, and 50nm in axial direction. In this scale, we can observe fine structure of cells, the distribution of proteins, and so on, which can not be identified by conventional microscope.
As a newcomer of super-resolution imaging, how to quickly master the skills and apply to my own project?
:First of all, super-resolution imaging is unique for its imaging mode, while for sample preparation part is almost the same when users doing general immunofluorescence staining procedures. What’s more, fluorescent markers Alexa647, Alexa750 are specially used in super-resolution imaging. For users, any project which includes fluorescence microscopy imaging is likely to dispose of a need for super-resolution imaging. Animal and plant cells, tissues, fungi, viruses and even DNA and RNA are all involved in the field of super-resolution imaging. If you choose our imaging service, you could take the samples to the company personally, or simply send us by mail after you finish the sample preparation part. Skilled technicians from NBI will help you to image the samples, along with the results and analysis report. The whole process does not involve users operation. If universities or research institutes choose to buy our system, NBI will provide hardware formation, software installation and a series of training process towards customers. NBI would help the customers to handle and operate the system skillfully once we deliver our products.
What is the format of the results? can I use open sourse software to analyse the data? or what kind of data analyse tool can you provide?
The format of files we got in SRiS imaging is tif raw-images and analysis results in csv, which we can be analyzed using ImageJ. Our customized products can provide users for their required type of data, and we will develop the product to be compatible with the mainstream open source analysis software.
How long does it take to take one SR image, and what is the size of the SR image data??
It will take several minutes for one SR image, and the size of SR image data is dozens of megabyte.
How to prepare SR sample?
SIn SRiS imaging, the sample need to be immunofluorescent staining, it is also possible to use samples with fluorescent proteins. It should be noted that when imaging with SRiS, specific class of fluorescent dyes (Alexa 647, Alexa 750, etc) and imaging buffer with particular formula should be used. NBI will guide the customer to prepare the sample and buffer.
If my samples contain endogenous GFP, RFP fluorescent proteins, can I use them to do the super-resolution imaging? How to achieve?
Yes. Alexa 647 or Alexa 750 labeled GFP nanobody can be used to directly label endogenous GFP or RFP proteins.
I alway use confocal microscope, may i use the sample in super resolution system?
In SRiS imaging, the sample need to be immunofluorescent staining, it is also possible to use samples with fluorescent proteins. It should be noted that when imaging with SRiS, specific class of fluorescent dyes (Alexa 647, Alexa 750, etc) and imaging buffer with particular formula should be used. So some sample for confocal can be used in SRiS imaging.
Why does the fluorescent dyes for super-resolution imaging is limited? Why does the commonly used channels in Confocal Microscope, such as 488 and 561 channels are not available in super-resolution imaging?
The blinking property of certain dyes is required in SRiS imaging, so not all fluorescent dyes can be used in super-resolution imaging.
How can I ship the sample to your facility? Is it feasible if your facility is not local?
Customers who are close to our company or imaging service center can choose to take the samples in person and take images. Customers who are far away from our company or imaging service centers can put samples and reagents in the ice box and send us by post. For the details of the mailing address and the mailing process, please contact NBI staffs for confirmation beforehand.
Compared with other companies of similar products, what advantages does your company have?
1) We have independently developed super-resolution imaging system. First, SRiS can do simultaneously dual-channel imaging, the buffer formula is optimized after rounds of test, so balanced results can be achieved in dual-channel. Second, active-locking system can make sure the drift of sample is within a few nanometers, the environment will not affect super-resolution imaging.
2) Biological basis and services: our biological team is lead by MIT postdoc, who have serviced many teams of well-known universities in Hong Kong and mainland China, so they have accumulated lots of experience.
3) We provide after-sales services, upgrade services, imaging services, and so on, which will be introduced in detail in the following questions.
What is your plan about training customers to operate SR imaging system.
Customers who choose the imaging service do not need to operate the instrument by themselves. Professional technicians in NBI are responsible for imaging your samples. If you choose to purchase the equipment, NBI staff will carry out a series of training, which includes: how to make imaging buffer, sample preparation, hardware and software operating procedures, imaging and data-analysis, etc. NBI promise that, users would handle and operate the system skillfully at the same time they bought our products. After training, customers can operate the super-resolution imaging system independently.
How about after-sales service?
After-sales service: the difference between super-resolution microscope and conventional microscope is that professional background is needed to get the best imaging results. Based on this, NBI is particularly concerned about after-sales service of our product. We will provide several round of training after the installation and testing of our product, and we will provide targeted guidance for our customers.
what is your super resolution service?
Due to high cost of super-resolution imaging equipment, not all research institutions can have their own equipment. So we can provide imaging service for these groups. First we will communicate with customers and guide them to prepare the sample, which can be mailed to NBI, and do super-resolution imaging on SRiS.
we have got a microscope in our lab, can you help upgrade to super resolution? and how?
Due to high cost of super-resolution imaging equipment, not all research institutions can have their own equipment. So we can help these groups to upgrade their conventional microscope to super-resolution microscope.