Nucleus/Nucleolus Imaging Services
Nucleolus is usually a single or multiple homogeneous spherical body, which is the most prominent compartment in the nucleus. Its main function is to coordinate the production and assembly of ribosomes, and participate in the regulation of cell cycle, apoptosis, DNA processing and DNA damage response. All these processes are directly related to human health. Therefore, the visual analysis of nucleolar morphology and localization is very important for the treatment of diseases.
Figure 1. Detecting three different classes of nucleolar morphology using NCL and FBL staining (Martin R M, et al. 2016).
Nucleolus Imaging Analysis
Defining nucleolar composition and activity under various growth conditions is a prerequisite for understanding its role in different aspects of cell physiology. At present, immunofluorescence staining with nucleolar protein-specific antibodies is a common tool for visual analysis of nucleolus, but it is very time-consuming and only suitable for fixed cells. Due to the wide range of nucleolar morphology observed, it is difficult to develop high-throughput image analysis methods to measure nucleolar changes.
CD BioSciences can capture different forms of nucleoli through high-content imaging, generate a large number of image data sets, then perform image analysis through computer software, and classify individual cell responses based on these parameters, so as to study many organisms related to nucleoli process.
Nucleolus Imaging Analysis Workflow
The following are the specific experimental steps of nucleolus imaging analysis:
Cell culture
The target cells are cultured in a growth medium. To subculture the cells, aspirate the medium and wash the cell monolayer once with dPBS, detach it with TrypsinEDTA, and then neutralize it with growth medium.
Step 1
Immunofluorescence analysis
After inoculating the target cells into a 384-well optical microplate for culture, aspirate the medium from the well and replace it with a growth medium containing a different concentration of the drug, then stain with a stain and wash the cells twice with dPBS.
Step 2
Image acquisition
Acquire images on a high-content screening system. The image was captured using the objective lens and the binning parameter set to 2 in the confocal mode. For each channel, a total of 36 fields of view per well are collected for analysis.
Step 3
High-content image analysis
We use image analysis software to identify and classify large-scale data sets, and generate readable analysis reports according to customer needs.
Step 4Delivery
Number of nucleoli for a specific type of abnormality
Linear classifier scatter plots
Intensity characteristics of effective nuclear staining and intensity median/average ratio
Other data that you need
Our Advantages
- High-throughput, high-resolution image acquisition
- Visual analysis of nucleolar morphology changes
- Provide a simple high-content image analysis algorithm
- Experienced scientists provide experimental consultation
- Reasonable price and short turnaround time
CD BioSciences has a professional team and advanced imaging equipment. The entire process of nucleolus imaging analysis is operated by experienced technicians to ensure the accuracy of the experiment. If you have any needs, please feel free to contact us.
- He J S, Soo P, Evers M, et al. High-content imaging approaches to quantitate stress-induced changes in nucleolar morphology[J]. Assay and drug development technologies, 2018, 16(6): 320-332.
- Martin R M, Herce H D, Ludwig A K, et al. Visualization of the nucleolus in living cells with cell-penetrating fluorescent peptides[M]//The Nucleolus. Humana Press, New York, NY, 2016: 71-82.
*If your organization requires the signing of a confidentiality agreement, please contact us by email.
Please note: Our services can only be used for research purposes. Do not use in diagnostic or therapeutic procedures!