Multifocal multiphoton microscopy(MMM)has recently become an important tool in biomedicine for performing three-dimensional fastfluorescence imaging.Using various beamsplitting techniques,MMM splits the near-infrared ...Multifocal multiphoton microscopy(MMM)has recently become an important tool in biomedicine for performing three-dimensional fastfluorescence imaging.Using various beamsplitting techniques,MMM splits the near-infrared laser beam into multiple beamlets and produces a multifocal array on the sample for parallel multiphoton excitation and then recordsfluorescence signal from all foci simultaneously with an area array detector,which significantly improves the imaging speed of multiphoton microscopy and allows for high efficiency in use of the excitation light.In this paper,we discuss the features of several MMM setups using different beamsplitting devices,including a Nipkow spinning disk,a microlens array,a set of beamsplitting mirrors,or a diffractive optical element(DOE).In particular,we present our recent work on the development of an MMM using a spatial light modulator(SLM).展开更多
Multifocal multiphoton microscopy(MMM)has greatly improved the utilization of excitationlight and imaging speed due to parallel multiphoton excitation of the samples and simultaneousdetection of the signals,which allo...Multifocal multiphoton microscopy(MMM)has greatly improved the utilization of excitationlight and imaging speed due to parallel multiphoton excitation of the samples and simultaneousdetection of the signals,which allows it to perform three-dimensional fast fuorescence imaging.Stochastic scanming can provide continuous,uniform and high-speed excitation of the sample,which makes it a suitable scanning scheme for MMM.In this paper,the graphical programminglanguage,LabVIEW is used to achieve stochastic scanning of the two-dimensional galvo scanmers by using white noise signals to control the a and y mirrors independently.Moreover,thestochastic scanning process is simulated by using Monte Carlo method.Our results show that MMM can avoid oversampling or subsampling in the scanning area and meet the requirements of uniform sampling by stochastically scanning the individual units of the N×N foci array.Therefore,continuous and umiform scaning in the whole field of view is implemented.展开更多
基金This work has been partially supported by NIH(SC COBRE P20RR021949 and Career Award 1k25hl088262-01)NSF(MRI CBET-0923311 and SC EPSCoR RII EPS-0903795 through SC GEAR program)+3 种基金The National Natural Science Foundation of China(31171372,61078067)Guangdong Province Science and Technology Project(2010B060300002)Shenzhen University Application Technology Development Project(201136,CXB201104220021A,JC201005250032A,200854)the Fundamental Research Funds for the Central Universities(K50510050006).
文摘Multifocal multiphoton microscopy(MMM)has recently become an important tool in biomedicine for performing three-dimensional fastfluorescence imaging.Using various beamsplitting techniques,MMM splits the near-infrared laser beam into multiple beamlets and produces a multifocal array on the sample for parallel multiphoton excitation and then recordsfluorescence signal from all foci simultaneously with an area array detector,which significantly improves the imaging speed of multiphoton microscopy and allows for high efficiency in use of the excitation light.In this paper,we discuss the features of several MMM setups using different beamsplitting devices,including a Nipkow spinning disk,a microlens array,a set of beamsplitting mirrors,or a diffractive optical element(DOE).In particular,we present our recent work on the development of an MMM using a spatial light modulator(SLM).
基金partially supported,by,the National Natural Science Foundation of China(11204226)the Fundamental Research Fundsfor,the Central Universities(K5051005006,K5051305002).
文摘Multifocal multiphoton microscopy(MMM)has greatly improved the utilization of excitationlight and imaging speed due to parallel multiphoton excitation of the samples and simultaneousdetection of the signals,which allows it to perform three-dimensional fast fuorescence imaging.Stochastic scanming can provide continuous,uniform and high-speed excitation of the sample,which makes it a suitable scanning scheme for MMM.In this paper,the graphical programminglanguage,LabVIEW is used to achieve stochastic scanning of the two-dimensional galvo scanmers by using white noise signals to control the a and y mirrors independently.Moreover,thestochastic scanning process is simulated by using Monte Carlo method.Our results show that MMM can avoid oversampling or subsampling in the scanning area and meet the requirements of uniform sampling by stochastically scanning the individual units of the N×N foci array.Therefore,continuous and umiform scaning in the whole field of view is implemented.
