Procedure for the optical acquisition of objects by means of a light raster microscope with line by line scanning

Abstract

Procedure for the image acquisition of objects by means of a light raster microscope with line by line scanning, whereas a scanning of the specimen for the creation of a specimen image occurs in scanning steps and the distance between at least two scanning steps is variably adjustable and at least a second scanning of the specimen occurs, during which the position of the scanning steps is shifted with regard to the scanning direction, whereas preferably a line by line scanning of the specimen is carried out.

1 . Process for image acquisition of objects using a light raster microscope with line by line scanning, comprising the steps of:
 (a) scanning of a specimen line by line in parallel scan lines in a scanning direction for the creation of a specimen image, wherein the distance between at least two scan lines is variably adjustable, and   (b) carrying out at least a second scanning of the specimen, during which the position of the scan lines is shifted with regard to the scanning direction.   
   
   
       2 . Process according to  claim 1 , further comprising the step of changing the proportion between spatial and temporal resolution of the microscope. 
   
   
       3 . Process for studying development processes, using the process of  claim 1 , wherein in step (a), the specimen exhibits dynamic processes on the level of united cell structures and whole organisms lasting from tenths of seconds up to hours. 
   
   
       4 . Process according to  claim 3 , wherein the specimen is living cells in a three-dimensional tissue group with markers, which exhibit manipulation related changes in color by laser illumination, in combination with living cells in a three-dimensional tissue group with very weak markers, and wherein the process further comprises the step of restricting confocality in favor of detection sensitivity. 
   
   
       5 . Process for studying intracellular transportation processes, using the process of  claim 1 , wherein the specimen is motile structures, with high speed in the area of hundredth of seconds in and wherein the scanning of steps (a) and (b) is carried out using FRAP with region of interest bleaches. 
   
   
       6 . Process for studying molecular and other sub-cellular interactions, using the process of  claim 1 , wherein the specimen is a very small structure with high velocity, and wherein the scanning of steps (a) and (b) is carried out using indirect techniques with region of interest bleaches for the resolution of submolecular structures. 
   
   
       7 . Process for studying fast signal transmitting procedures, using the process of  claim 1 , wherein the specimen exhibits neurophysiological processes with high temporal resolution.