High-speed confocal fluorescence imaging with a novel line scanning microscope.
Research in the life sciences increasingly involves the investigation of fast dynamic processes at the cellular and subcellular level. It requires tools to image complex systems with high temporal resolution in three-dimensional space. For this task, we introduce the concept of a fast fluorescence line scanner providing image acquisition speeds in excess of 100 frames per second at 512 x 512 pixels. Because the system preserves the capability for optical sectioning of confocal systems, it allows us to observe processes with three-dimensional resolution. We describe the principle of operation, the optical characteristics of the microscope, and cover several applications in particular from the field of cell and developmental biology. A commercial system based on the line scanning concept has been realized by Carl Zeiss (LSM 5 LIVE).
Wolleschensky
Ralf
R
Carl Zeiss MicroImaging GmbH, Carl-Zeiss-Promenade 10, 07745 Jena, Germany. Wolleschensky@zeiss.de
Zimmermann
Bernhard
B
Kempe
Michael
M
D000465:Algorithms; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007089:Image Enhancement / Q000295:instrumentation* / Q000379:methods; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D018613:Microscopy, Confocal / Q000295:instrumentation* / Q000379:methods; D008856:Microscopy, Fluorescence / Q000295:instrumentation* / Q000379:methods; D018715:Microscopy, Video / Q000295:instrumentation* / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity