共聚焦显微成像技术及图像处理 - CAS

1 2020 8 7 Leica DLS Leica SP8 Zeiss LSM 710 NLO Leica STED Rat Brain, Double labelling: Green: Neurons, Blue: Nuclei PMT Laser Rat Brain, Double labelling: Green: Neurons, Blue: Nuclei Pinhole Laser Sample PMT Pinhole XY scanning Z-Drive d NA Laser 405nm 458nm/488nm/514nm 561nm 594nm 633nm Laser 405nm 448nm 488nm 552nm 638nm Laser 405nm 458nm/476nm/488nm/496nm/514nm WIL 470nm-670nm Laser 690nm-1064nm 458nm/488nm/514nm 561nm 633nm 90 PMT PMT 1 2 3

2 Nm PDE HyD : Pinhole Pinhole Pinhole Z-stack 3D Pinhole

3 Z-stack 3D : xyt xyzt xt Ca2+ pH Pinhole Z-stack 3D : xyt xyzt xt Ca2+ pH Tilescan Pinhole Z-stack 3D : xyt xyzt xt Ca2+ pH Tilescan Mark and Find Pinhole Z-stack 3D : xyt xyzt xt Ca2+ pH Tilescan Mark and Find Pinhole Z-stack 3D.

4 Xyt xyzt xt Ca2+ pH Tilescan Mark and Find PA-GFP and HcRed cotransfected NRK cells paGFP BMP signaling is required for pharyngeal pouch progenitor specification in zebrafish ( ) COS cells expressing p75-GFP Courtesy Prof. Dr. Ralf Jacob, University Marburg 2020 EMBL Matteo Rauzi and Lars Hufnagel Scientific Reports 1025nm Crop Merge Project Huygens Lightning 3D -Crop -Merge -Projection - - Median Gaussian Blur -Despeckle -Huygens -3D - - ROI 3D - Colocalization Rate Scatter Plot Plot profile Manders' Colocalization Coefficients (MCC)

5 TEL 180 4056 1678 MAIL TEL: 136 3865 1076 MAIL TEL 135 4506 8558 MAIL frap FRET STED XY 120nm