title: The use of a laser for correlating light and electron microscopic images in thick tissue specimens.
abstract: Correlating images between light and electron microscopy is difficult especially in tissue specimens with a substantial z-dimension. To facilitate correlated light and electron microscopy (CLEM) in thick tissue specimens, we describe a basic method of using a femto-pulsed near-infrared laser to burn precise three-dimensional fiducial markers that circumscribe cells or regions of interest for easy identification between imaging methods. This rapid and reliable approach permits traditional fixation and avoids the use of electron-dense labeling methods that can obscure ultrastructural details. The versatility of the technique permits CLEM in a variety of tissue specimens to allow interpretation of highly resolved ultrastructural data in the more macroscopic and potentially dynamic context of light microscopy.
authors.affiliation: Department of Physiology, Indiana University School of Medicine-Muncie, Muncie, Indiana, USA.
authors.forename: Derron
authors.initials: D
authors.lastname: Bishop
authors.affiliation: Research Unit Therapy Development, Institute of Clinical Neuroimmunology, Ludwig-Maximilian University, Munich, Nordallee, Germany.
authors.forename: Ivana
authors.initials: I
authors.lastname: Nikic
authors.affiliation: Research Unit Therapy Development, Institute of Clinical Neuroimmunology, Ludwig-Maximilian University, Munich, Nordallee, Germany; Munich Center for Systems Neurology (SyNergy), Nordallee, Munich, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Kerschensteiner
authors.affiliation: Munich Center for Systems Neurology (SyNergy), Nordallee, Munich, Germany; Institute of Neuronal Cell Biology, Technische Universität München, Nordallee, Munich, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Misgeld
mesh_terms: D000818:Animals; D002540:Cerebral Cortex / Q000648:ultrastructure; D021621:Imaging, Three-Dimensional; D008822:Mice, Transgenic; D018613:Microscopy, Confocal / Q000379:methods; D046529:Microscopy, Electron, Transmission / Q000379:methods; D008867:Microtomy
keywords: Confocal microscopy; Electron microscopy; In vivo; Multiphoton microscopy; Serial section reconstruction; Technique