Project
320-Spratt-dMetRaman
Title
Stimulated Raman images of deuterated methionine incorporated into adult and larval Drosophila
Description
Introduction: Visualizing small individual biomolecules at subcellular resolution in live cells and tissues can provide valuable insights into metabolic activity in heterogeneous cells, but is challenging. Methods: Here, the authors used stimulated Raman scattering (SRS) microscopy to image deuterated methionine (d-Met) incorporated into Drosophila tissues in vivo. Results: Their results demonstrate that SRS can detect a range of previously uncharacterized cell-to-cell differences in d-Met distribution within a tissue at the subcellular level. Discussion: These results demonstrate the potential of SRS microscopy for metabolic imaging of less abundant but important amino acids such as methionine in tissue.
Funding
This work was supported in part by Japan Society for the Promotion of Science KAKENHI under JP20H02650, JP20H05725, JP20H05726, and JP21J00452; in part by Japan Science and Technology Agency Core Research for Evolutional Science and Technology (CREST) under Grant JPMJCR 1872; in part by Nakatani Foundation Grant for Technology Development Research, and in part by Quantum Leap Flagship Program of the Ministry of Education, Culture, Sports, Science and Technology (MEXT) under Grant JPMXS0118067246.
Title
Stimulated Raman scattering of dMet in Drosophila larval fat body.
Description
Stimulated Raman scattering images of deuterated methionine (dMet) uptake in Drosophila larval fat body. The Raman peak of dMet is 2130cm^{-1}. The images of LysoTracker fluorescence in the same area is also reposited.
Description
The dissected fat body from the third instar larvae of Drosophila which was incorporated deuterated methionine (dMet).
Extrinsic variables
Flies were transferred to a modified version of holidic medium supplemented with deuterated methionine (Cambridge Isotope Laboratories: DLM-6797–0.1) instead of methionine, for up to 48 h prior to dissection.
Gene ontology: Biological processes
Gene ontology: Cellular components
Gene ontology: Molecular functions
Microscopy types
inelastic scattering of photons (Raman scattering) (
FBbi:00000589)
(stimulated Raman spectroscopy)
Detection methods
Home-made detector with a Si photodiode (Hamamatsu S3399), electric filters, amplifers, and a lock-in amplifier.
Visualization methods
deuterated methionine (d8-Met)
Illumination methods
Home-made detector with a Si photodiode (Hamamatsu S3399), electric filters, amplifers, and a lock-in amplifier.
Contrast enhancing methods
Resolution enhancing methods
Sample preparation methods
living tissue
Light source
Picosecond Ti:sapphire laser (Coherent, Mira900D) and synchronized Yb fiber laser
Detector
Home-made detector with a Si photodiode (Hamamatsu S3399), electric filters, amplifers, and a lock-in amplifier.
Objective
Olympus 60x, NA1.2