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    • 4. 发明申请
    • CONFOCAL MICROSCOPE AND RELATED PROCESS OF ACQUIRING AND PROCESSING IMAGES
    • 共焦显微镜及其获取和处理图像的相关过程
    • WO2016199025A1
    • 2016-12-15
    • PCT/IB2016/053344
    • 2016-06-08
    • CRESTOPTICS S.R.L.CONSIGLIO NAZIONALE DELLE RICERCHE
    • VITULANO, DomenicoBRUNI, VittoriaRICCO, VincenzoSANTINELLI, Andrea
    • G02B21/00G02B27/58
    • G02B21/008G02B21/0044G02B21/0072G02B27/58
    • Process of acquiring and processing images in confocal microscopy, comprising the following steps: A. acquiring (500, 510; 520, 530) a first overall image A k-1 of a plane of a sample (160) scanning the same with a first excitation light beam, obtained by making a basic light beam pass through first pinholes (1255; 1255') and transmitted to an optics (155) of a confocal microscope (100) that is focused on the plane of the sample (160), and acquiring a second overall image Ak of the plane of the sample (160) scanning the same with a second excitation light beam, obtained by making said basic light beam pass through second pinholes (1265; 1265') having same shape and size (L 2 ; L a2 , L b2 ) larger than the first pinholes (1255; 1255'); B. calculating (540) an estimate g of the PSF of the optics (155) of the microscope (100), through an inverse Fourier transform of formula (I): where  k -1 (ω) is the Fourier transform of the acquired first overall image A k-1 and  k (ω) is the Fourier transform of the acquired second overall image A k ; and C. deconvolving (550) the acquired second overall image A k using the estimate g of the PSF of the optics (155) of the microscope (100) through a main deconvolution algorithm, obtaining the resulting image / of the entire plane of the sample (160): I = A k • g.
    • 在共聚焦显微镜中获取和处理图像的过程包括以下步骤:A.采集(500,510,520,530)第一个整体图像Ak-1,其中第一个整体图像用第一个激发扫描的样品(160)的平面 通过使基本光束通过第一针孔(1255; 1255')并被传输到聚焦在样品(160)的平面上的共聚焦显微镜(100)的光学器件(155)而获得的光束,并获取 通过使所述基本光束通过具有相同形状和尺寸(L2; La2)的第二针孔(1265; 1265')而获得的样品(160)的平面的第二整体图像Ak与第二激发光束扫描相同 ,Lb2)大于第一针孔(1255; 1255'); B.通过公式(I)的傅立叶逆变换计算(540)显微镜(100)的光学器件(155)的PSF的估计值:其中kk(ω)是所获取的傅立叶变换 第一整体图像Ak-1和k(ω)是所获取的第二整体图像Ak的傅立叶变换; 和C.通过主去卷积算法使用显微镜(100)的光学器件(155)的PSF的估计值g来获取的第二整体图像Ak解卷积(550),获得样品的整个平面的所得图像 (160):I = Ak·g。