Multiphoton Laser Scanning Microscope FV1000MPE : Applications
Zebrafish
Transgenic zebrafish with cell membranes labeled with CFP.
CFP is shown in cyan and YFP in green.
Specimens provided by:
Dr. Rachel O Wong, Mr. Philip Williams,
Dept. Biological Structure, University of Washington
Silkworm
3-dimensionally constructed image of cGMP-containing cells marked with CY3 running along the antenna nerve of the silkworm.
200 µm projection image.
Specimens provided by:
Hitoshi Aonuma
Research Institute for Electronic Science, Hokkaido University, Japan
Mouse/Rat brain
3-dimensionally constructed images of neurons expressing EYFP in the cerebral neocortex of a mouse under anesthesia.
Cross-sectional images down to 0.7 mm from the surface can be observed after attachment of a special adapter to the specimen.
Objective: LUMPlanFL 60xW/IR
Specimens provided by:
Hiroaki Waki, Tomomi Nemoto, and Junichi Nabekura
National Institute for Physiological Sciences,
National Institutes of Natural Sciences, Japan
Calcium signal of a single dendritic spine examined by multiphoton uncaging and fluorescence
a) Stacked fluorescent image of dendritic spines in the hippocampus (excitation of 830 nm). Whole-cell
recording was performed, and Alexa 594 and the calcium indicator OGB-5N were injected. At the head
of the single spine (red), multiphoton uncaging of caged glutamate was performed and glutamate was
administered (excitation of 720 nm). A line scan was performed on the line (the line linking the 2
triangles) from the head of this single spine toward the dendritic trunk.
b), c) Simultaneous line scanning for Alexa 594 and OGB-5N.
d) Calcium concentration determined from the fluorescence emission ratios of OGB-5N and Alexa 594.
e) Changes in calcium concentration at the head of the spine (H, black), changes in calcium
concentration at the dendritic trunk (D, red), current from whole-cell recorded NMDA receptors (INMDA).
Calcium flow into the trunk via NMDA receptors at the head of the spine is apparent from these
observations.
Reprinted from Noguchi et al. Neuron 46(2005)609-622.
Jun Noguchi, Haruo Kasai
Center for Disease Biology and Integrative Medicine, Faculty of Medicine,
University of Tokyo
Second Harmonic Generation imaging of neurons.
A: SHG image of neurons in dissociated culture from the mouse cerebral
cortex. After FM4-64 was injected to neurons, the cells were irradiated
with a femtosecond laser at 950 nm and the SHG signal at 475 nm was
detected with the transmitted light detector.
B: Zoomed fragment (5X) of the spacimen in the yellow box in image A.
As it is apparent, spines protruding from dendrites can be observed with
SHG.
C: SHG and multiphoton images have been superimposed.
Image data provided by:
Mutsuo Nuriya, PhD, Masato Yasui, MD, PhD
Department of Pharmacology School of Medicine, Keio University
