The workflow is as follows:

Laser focusing: A laser beam with good monochromaticity is focused onto a specific focal plane of the sample, exciting the fluorescent probe to produce fluorescence.

Pinhole screening: Only fluorescence signals on the focal plane can pass through the pinhole in front of the detector, and stray light from non focal planes will be blocked by the pinhole, resulting in higher image contrast.

Scanning imaging: The laser beam scans the entire focal plane point by point and line by line, and the computer collects the signal and reconstructs a two-dimensional image. By moving the sample along the Z-axis, a series of slices of different depths can be obtained, and then a three-dimensional image of the sample can be reconstructed.

Laser confocal microscopeThe main characteristics are:

High resolution and contrast: effectively eliminates non focal plane interference, with a lateral resolution of about 120-200nm, and imaging quality far exceeding traditional wide field fluorescence microscopes.

Three dimensional imaging: Optical slicing of the sample can be performed without the need for physical slicing, and the three-dimensional structure can be reconstructed through software to clearly display the spatial form of the sample.

Multicolor fluorescence imaging: can be equipped with lasers of multiple wavelengths to simultaneously excite different fluorescent probes in the sample, distinguishing different target structures on the same image.

Dynamic observation of live cells: With minimal damage to the sample, it can track the dynamic physiological processes inside live cells in real-time for a long time under temperature and humidity control environments.

Powerful analytical capabilities: The supporting software can perform various data processing such as image enhancement, cell counting, fluorescence intensity quantification, co localization analysis, etc.