Multi-channel Frequencies Division Multiplexed Fluorescence Confocal Microscopy

With the continuous improvement of science and technology, industrialmanufacturing technologies and computer image processing technology, the detectionaccuracy and resolution of laser scanning confocal microscope (LSCM) are beingconstantly improved and increased. Today, the requirements on the detection rate ofthe microscope become increasingly higher in the fields of cell biology, biomedical,and medical testing. However, most available LSCMs in the market currently haveonly one or two channels. Therefore it is difficult to meet the increasing demands ofdevelopment of fields mentioned above. The detection rate has become a "bottleneck"which limits the improvement of LSCM.To solve the problem of how to improve the detection rate of LSCM, our grouppresents a frequencies division multiplexed fluorescence confocal microscopy(FDMFCM) which is based on H-PDLC Bragg grating. With high diffractionefficiency of H-PDLC Bragg grating, the microscopy is able to improve the detectionrate of LSCM by increasing number of detecting channel. In the microscopy, multipledetecting channels are achieved with light chopper array fabricated with severalH-PDLC Bragg gratings. This particular chopper functions as both spectroscopes andmodulators in the microscope system, thus it is the key to achieve multi-channelparallel detection.The four-channel FDMFCM system is experimentally set up with405nm laser asthe excitation light. Images of mouse neural hippocampus cells sample are takenand intensity information of the fluorescence signal excited from the sample, aresuccessfully detected. Experiments show that the proposed FDMFCM is feasible forenhancement of detection rate. This technology provides a novel, useful and efficientmethod for areas such as medicine, biological cell research.