Zeiss LSM 510 Operating Manual

Also see for LSM 510 Inverted: Quick guide Operating manual User guide User manual Brief Operating Manual

INTRODUCTION TO LASER SCANNING MICROSCOPYThree-Dimensional Presentation of LSM Image Stacks LSM 5103-4 B 40-051 e 07/98 In order to obtain an image of the selected object plane as a whole, it is necessary to scan the objectplane in a point-by-point, line-by-line raster by means of an XY light deflection system. The detectors -as a rule, photomultipliers - convert the optical information into electric signals. This allows the imageof any object plane to be generated and stored within less than a second. By a defined focusing(Z axis) movement it is possible to look at any object plane of interest. By scanning a succession ofobject planes in a specimen, a stack of slice images can be produced.This way, the LSM technique in conjunction with ICS optics (Infinity Color-Corrected System) has broughtdecisive improvements over conventional microscopy in terms of resolving power and confocal depthcontrast:Object features in the order of 0.2 m can be resolved, and height differences of less than0.1 m made visible, without the use of interference methods.3.2 Three-Dimensional Presentations of LSM Image StacksOne of the advantages of the LSM technique is that it can present structures in three dimensions. Thisopens up many ways to process images. Outlined below are some of the possible methods to extractspatial information from stacks of slice images. GalleryThe simplest presentation of 3D information is a gallery showing the individual slice images (sections)of a stack arranged side by side, with each slice apart from the next by a defined, selectable intervalon the Z axis. Virtually infinite depth of focusThe entire set of data can be imaged as a single projection. The computer establishes an imagecomposed of all in-focus optical sections. The image produced by this so-called composite methodhas a virtually infinite depth of focus, since the result is made up of information from in-focus planesonly. Rotary animationA sequence of projections is computed, with the specimen being apparently rotated by a certainangle from image to image, for example by a full turn about an axis. If such a sequence is displayedon the monitor screen in rapid succession, the visual effect is that of a rotating three-dimensionalobject. Stereo image pairsThe computer establishes a pair of images corresponding to those we see with the right and the lefteye, respectively. The two images forming the stereo pair can be shown on the monitor side by side.They can be seen as a 3D image with suitable optical aids. Another possibility is to present bothimages in registration, with one image in the red channel and the other in the green one (anaglyph).Viewed through red and green color filters in a spectacle frame, which only pass the image intendedfor the respective eye, the two images form a 3D image in the brain Color-coded height slicesEach level, i.e. each slice is assigned a different color. For direct evaluation, a color scale is shown,indicating the actual height above the bottom slice.