Measurement of the Transfection Efficiency of Cultivated Cells

Capturing clear images without fluorescence blurring

Transfection is the process of introducing nucleic acids into animal cells. It is used to make cells introduce specific genes to express a protein of interest.

One of the results of this process is that transfection efficiency can be analyzed. Hence, this process is vital to current medical research.

The success or failure of a gene transfer relies on the optimization of the transfer conditions. As such, various researchers have repeatedly carried out trial and error in search of the optimal process.

There is a wide range of processes, but they can be classified into the following three main types: (1) chemical processes, (2) biological processes, and (3) physical processes. Each process has its own merits and demerits, so it is necessary to select the appropriate process to match the needs of the experiment and the cell type.

After selecting the optimal process, prepare the soundness and survival rate of the cell system, the cell density, and other such components required to ensure the transfection is successful, and then start the experiment. Among these components, although it is extremely important to accurately measure the transfection efficiency, many people actually have a great deal of trouble doing so.

One general method for measuring the transfection efficiency is to use a fluorescence microscope. The transfection efficiency is measured by counting the total number of observed cells and the number of cells that express fluorescence and scoring these values. However, when using conventional fluorescence microscopes, this approach posed various problems and was often very difficult.

First, to observe the fluorescence it was necessary to take the specimen to a darkroom and carry out the experiment there, thereby resulting in extremely poor operability. Furthermore, it was difficult to accurately extract the outlines of cells, which hindered the accurate counting of the number of cells.

Photobleaching due to fluorescence observation was a headache for many researchers. The health of the cells is extremely important, so a method that does not weaken the cells was required.

In addition, to actually count and analyze the number of cells, software separate from the fluorescence microscope system was required, thereby leading to many customers noting that it was difficult to perform smooth analyses.

Counting cells using a phase contrast image
Counting cells using a phase contrast image
Counting expression events (fluorescent points) using cells as mask areas
Counting expression events (fluorescent points) using cells as mask areas
Counting expression events (fluorescent points) using cells as mask areas
Cell
903
Expression
89
Efficiency
9.9%

Objective lens: CFI Plan Fluor DL 10x

Using the All-in-One Fluorescence Microscope BZ-X800