Golgi Structure Imaging Services
The Golgi apparatus is a highly polar organelle formed by piles of several flat vesicles. It is often distributed between the endoplasmic reticulum and the cell membrane in an arcuate or hemispherical shape. The Golgi apparatus is a dynamic organelle that regulates the transport of vesicles. Although cell transport requires positive changes in the Golgi membrane, these are not accompanied by changes in the general Golgi structure. However, cellular processes such as mitosis, apoptosis and migration require the fragmentation of the Golgi apparatus. Therefore, the visual analysis of the Golgi apparatus is very important for biological research.
Figure 1. Golgi apparatus structure.
Golgi Structure Imaging Analysis
At present, the changes in the structure of the Golgi are often studied by basic immunofluorescence and quantified by manual and subjective classification of the Golgi structure in 100-500 stained cells. There are several other high-throughput methods, but these methods are either complicated or do not provide sufficient morphological information. Therefore, it is necessary to develop a simple and informative Golgi research method.
CD BioSciences can detect and quantify Golgi structure changes in a non-subjective manner through imaging flow cytometry (IFC). We use the unique ability of IFC to quickly acquire tens of thousands of high-quality multispectral images, and then quantify Golgi fragmentation based on Golgi protein staining. The method is completed in an automated and unbiased manner, without laborious manual image acquisition, and avoids the variability between users in image analysis.
Golgi Structure Imaging Analysis Workflow
The following are the specific experimental steps of Golgi structure imaging analysis:
Step 1: Cell culture
The HeLa cells were cultured in a medium, and the cells were kept in a humidified atmosphere of 95% air and 5% CO2 at 37°C.
Step 2: Cell synchronization
HeLa cells were treated with 2.5 mM thymidine in DMSO, washed twice with PBS, grown in a regular medium for 8 h, treated with 2.5 mM thymidine for 16 h, and then washed with PBS.
Step 3: Apoptosis Induction and treatment
Cycloheximide was used to induce HeLa cell apoptosis for up to 6 hours, and Brefeldin A at different concentrations was used to induce Golgi rupture.
Step 4: Multispectral IFC analysis
After mixing the cells with primary antibodies, secondary antibodies, etc., use multispectral imaging flow cytometry for imaging, and use image analysis software for Golgi structure analysis.
Delivery
Percentage of cells in the complete, partially and fully fragmented Golgi apparatus at each stage
The area (A), width (W) and height (H) of Golgi marker staining
RD value and other data that you need
Our Advantages
- High-throughput, high-resolution image acquisition
- Simple and informative
- Visual analysis of Golgi structure changes
- 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 Golgi structure 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. We will provide you with personalized imaging analysis solutions and highly reproducible experimental data.
- Wortzel I, Koifman G, Rotter V, et al. High throughput analysis of Golgi structure by imaging flow cytometry[J]. Scientific reports, 2017, 7(1): 1-11.
- Lowe M. Structural organization of the Golgi apparatus[J]. Current opinion in cell biology, 2011, 23(1): 85-93.
*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!