title: Eyecup scope--optical recordings of light stimulus-evoked fluorescence signals in the retina.
abstract: Dendritic signals play an essential role in processing visual information in the retina. To study them in neurites too small for electrical recording, we developed an instrument that combines a multi-photon (MP) microscope with a through-the-objective high-resolution visual stimulator. An upright microscope was designed that uses the objective lens for both MP imaging and delivery of visual stimuli to functionally intact retinal explants or eyecup preparations. The stimulator consists of a miniature liquid-crystal-on-silicon display coupled into the optical path of an infrared-excitation laser-scanning microscope. A pair of custom-made dichroic filters allows light from the excitation laser and three spectral bands ('colors') from the stimulator to reach the retina, leaving two intermediate bands for fluorescence imaging. Special optics allow displacement of the stimulator focus relative to the imaging focus. Spatially resolved changes in calcium-indicator fluorescence in response to visual stimuli were recorded in dendrites of different types of mammalian retinal neurons.
authors.affiliation: Department of Biomedical Optics, Max-Planck-Institute for Medical Research, Jahnstr. 29, 69120 Heidelberg, Germany. thomas.euler@mpimf-heidelberg.mpg.de
authors.forename: Thomas
authors.initials: T
authors.lastname: Euler
authors.forename: Susanne E
authors.initials: SE
authors.lastname: Hausselt
authors.forename: David J
authors.initials: DJ
authors.lastname: Margolis
authors.forename: Tobias
authors.initials: T
authors.lastname: Breuninger
authors.forename: Xavier
authors.initials: X
authors.lastname: Castell
authors.forename: Peter B
authors.initials: PB
authors.lastname: Detwiler
authors.forename: Winfried
authors.initials: W
authors.lastname: Denk
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D003712:Dendrites / Q000502:physiology / Q000528:radiation effects; D007834:Lasers; D008433:Mathematics; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation* / Q000379:methods; D009474:Neurons / Q000502:physiology; D055095:Optics and Photonics; D010775:Photic Stimulation; D020419:Photoreceptor Cells, Vertebrate / Q000528:radiation effects; D011817:Rabbits; D012160:Retina / Q000502:physiology* / Q000528:radiation effects; D012984:Software
chemical_list: D002118:Calcium