Introduction of main components and functions of laser confocal microscope
1. Lighting pinhole Function: Make the laser pass through the illumination pinhole to form a point light source, which has the unique advantages of strong directivity, small divergence, high brightness, high spatial and temporal coherence and plane polarization excitation. And form a confocal device with that pinhole and focal plane of the detector.
2. Beam splitter Function: separate the excited fluorescence of the sample from other non-signal light.
3. objective lens
4. Focal plane Function: The laser point light source irradiates the object to focus at the focal plane, and excites the fluorescent labeled sample to emit fluorescence, forming a focal spot. The spot is processed by a series of devices, such as objective lens and beam splitter, and focused at the illumination pinhole and the detector pinhole respectively. The meaning of confocal comes from this.
5. Detector pinhole Function: It acts as a spatial filter. * Obstruct the scattered light from the non-focusing plane and the scattered light from the non-focusing spot on the focusing plane to the maximum extent, so as to ensure that all the fluorescence signals received by the detector pinhole come from the focus position of the sample spot, so the diffraction focused spot on the sample and the imaging spot of the detector pinhole contain the same information (two points conjugate).
6. Photomultiplier tube (detector) Function: Receive the optical signal passing through the pinhole, convert it into electrical signal and transmit it to the computer, and a clear image of the whole focal plane will appear on the screen.
7. Laser: The development of confocal microscope technology is inseparable from the rapid development of lasers. We can choose different lasers according to the research needs. Such as ARUV (351.364 nm), HECD (442 nm), AR (457,488,514 nm), ARKR (488,568,647 nm) KR (568 nm), Hene (543 nm), Hene (633 nm) and so on. 8. Multi-fluorescence channels: there are multiple fluorescence channels to realize multiple labeling of samples at the same time.
