Endoplasmic Reticulum Imaging Services

Endoplasmic Reticulum Imaging Services

The endoplasmic reticulum (ER) is an organelle in eukaryotic cells that forms a network of interconnected flat, membrane-enclosed sacs or tubular structures. It comes in two forms: ribosome-bound membranes (rough endoplasmic reticulum) or unbound ribosome-bound membranes (smooth endoplasmic reticulum). The endoplasmic reticulum plays a central role in lipid and protein synthesis, protein folding, and calcium homeostasis. Its dysfunction is related to many diseases, which are usually manifested as changes in the network structure and organization. Therefore, it is very important to use live cell imaging technology to quantitatively describe the changes of this organelle.

Drug-induced ER reorganisation in U-2 OS cellsFigure 1. Drug-induced ER reorganization in U-2 OS cells (Garcia-Pardo M E, et al. 2021).

Endoplasmic Reticulum Imaging Analysis

Although the importance of ER in cell function and dysfunction is obvious, reliable and repeatable analytical tools to quantify ER conformation are very limited. Living cell imaging technology can display molecular events in real time. It is an image analysis method that can quantitatively describe the changes of this organelle.

CD BioSciences has constructed a high-content imaging analysis platform for automated analysis of ER structure. It uses an automated high-content screening microscope to capture cell images of various fluorescently-labeled ER cells and then perform quantitative analysis on them. High-content imaging enables researchers to detect changes in ER structure under different conditions, which helps to study this complex organelle in mammalian cells.

Endoplasmic Reticulum Imaging Analysis Workflow

The following are the specific experimental steps of endoplasmic reticulum imaging analysis:

Step 1: Cell culture

The stable expression target cells were inoculated into the culture medium and cultured in a humidified incubator containing 5% CO2 at 37 °C.

Step 2: Cell processing and staining

Inoculate cells in a 96-well optically transparent plate, then fix and stain the entire plate simultaneously, and treat it with different concentrations of compounds.

Step 3: Imaging sample preparation

After various treatments, the nucleus and plasma membrane were respectively counterstained in DMEM medium, and then PBS solution was used and stored in the solution for imaging.

Step 4: Auto confocal imaging

On the high content screening system, the region of interest is imaged by confocal imaging.

Step 5: Image analysis

Use high-content analysis software to analyze images, and perform quantitative analysis on rough ER and smooth ER.

Delivery

ER distribution in cells
Area of dense ER

The area of the polygonal area between the mesh elements
Morphology of RER and SER in cell population and other data

Our Advantages

  • Allows visualization of the ER structure
  • High-sensitivity optical system to capture high-quality pictures
  • 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 endoplasmic reticulum 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.

References
  1. Garcia-Pardo M E, Simpson J C, O'Sullivan N C. A novel automated image analysis pipeline for quantifying morphological changes to the endoplasmic reticulum in cultured human cells[J]. BMC bioinformatics, 2021, 22(1): 1-17.
  2. Bader C A, Sorvina A, Simpson P V, et al. Imaging nuclear, endoplasmic reticulum and plasma membrane events in real time[J]. FEBS letters, 2016, 590(18): 3051-3060.

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Please note: Our services can only be used for research purposes. Do not use in diagnostic or therapeutic procedures!

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