Simultaneous imaging of different focal planes in fluorescence microscopy for the study of cellular dynamics in three dimensions.
The imaging of cellular dynamics in three dimensions using a standard microscope is severely limited due to the fact that only one focal plane can be imaged at a given point in time. Here we present a modification of the classical microscope design with which two or more focal planes can be imaged simultaneously. This is achieved by a modification of the emission pathway of a standard microscope. The efficacy of the design is shown by imaging bead samples and an FcRn-green fluorescent protein expressing tubule that leaves a sorting endosome and subsequently exocytoses at the plasma membrane.
Prabhat
Prashant
P
Department of Electrical Engineering, University of Texas at Dallas, Richardson, TX 75083-0688, USA. prashant.prabhat@student.utdallas.edu
Ram
Sripad
S
Ward
E Sally
ES
Ober
Raimund J
RJ
D002460:Cell Line; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D015395:Histocompatibility Antigens Class I; D006801:Humans; D007089:Image Enhancement / Q000295:instrumentation*; D021621:Imaging, Three-Dimensional / Q000295:instrumentation* / Q000379:methods; D008856:Microscopy, Fluorescence / Q000295:instrumentation* / Q000379:methods; D018715:Microscopy, Video / Q000295:instrumentation* / Q000379:methods; D011961:Receptors, Fc / Q000378:metabolism*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D015395:Histocompatibility Antigens Class I; D011961:Receptors, Fc; C100688:Fc receptor, neonatal