Laser Scanning Confocal Microscope Imaging Data Processing Method Based on Integral Sampling
A confocal microscope and laser scanning technology, applied in microscopes, optics, optical components, etc., can solve problems such as operating frequency limitations, and achieve the effect of increasing amplitude, high sampling voltage, and improving signal-to-noise ratio.
Active Publication Date: 2017-05-31
SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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In the way of sampling the PMT output signal after analog integration within the pixel dwell time, the integrator must be cleared every time a point is scanned, that is, it involves charging, sampling, and discharging the integrator within a pixel time, although for The image signal-to-noise ratio enhancement effect is strong, but limited by the operating frequency of the switching device in its mechanism and the voltage change rate of the operational amplifier, its final operating frequency is also limited
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[0033] The present invention will be further described in detail below in conjunction with the accompanying drawings, so that those skilled in the art can implement it with reference to the description.
[0034] To achieve the above object, the present invention is achieved through the following technical solutions:
[0035] Laser Scanning Confocal Microscope Imaging Data Processing Method Based on Integral Sampling, see figure 1 , including the following steps:
[0036] Step 1) collecting the signal data of the sample to be imaged pixel by pixel with the photomultiplier tube in the laser scanning confocal microscope;
[0037] Step 2) first integrate the signal data obtained by scanning the first pixel, and the integration time is from 0 to T to obtain the brightness value U(T) of the first pixel, where T is the scanning time of each pixel; At one point, the capacitor of the integrator is loaded with the voltage signal superimposed on the second pixel and the first pixel, wh...
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The invention discloses an LSCM (laser scanning confocal microscope) imaging data processing method based on integral sampling. The LSCM imaging data processing method based on the integral sampling includes following steps: 1) using a photomultiplier in an LSCM to collect signal data of a sample to be imaged per pixel; 2) using the photomultiplier to amplify the signal data of each pixel, which is obtained through scanning, and then using an integrator to perform integration on the signal data, wherein firstly, integrating the signal data and scanning time, which are obtained when the first pixel is scanned, then integrating the signal data and scanning time, which are obtained when the second pixel is scanned, and finally completing the integration until the signal data and scanning time, which are obtained when the Nth pixel is scanned, are integrated, and furthermore setting N to be large than or equal to 1; 3) using integration value obtained when the first pixel is scanned as brightness value of the first pixel, using difference value between integration value obtained when the second pixel is scanned and the integration value obtained when the first pixel is scanned as brightness value of the second pixel, and sequentially calculating difference value of integration value of each two adjacent pixels until obtaining brightness value of the Nth pixel. The LSCM imaging data processing method based on the integral sampling improves imaging quality of the LSCM.
Description
technical field [0001] The invention relates to a laser scanning confocal microscope imaging data processing method, in particular to a laser scanning confocal microscope imaging data acquisition and processing method based on integral sampling. Background technique [0002] Laser Scanning Confocal Microscopy (LSCM) is an effective technique for studying submicron microstructures. It is a necessary large-scale scientific research instrument for scientific and technological workers engaged in biomedical and material science research at home and abroad. [0003] In the LSCM system, the scanning galvanometer reflects the scanning spot, drives the scanning spot to move through the X-Y scanning mechanism and traverses the imaging area on the sample, and the fluorescent signal returned from the sample passes through the optical path and reaches the photomultiplier tube (Photomultiplier Tube, PMT), PMT The output is a continuous analog voltage signal within the sweep time. If the...
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IPC IPC(8): H04N5/341H04N5/353H04N5/357G02B21/36
Inventor 黄维张运海
Owner SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI