Raman microscopeFeatures
Molecular structure

Principle of Raman scattering

Observation of wavelength shift
originated from the interaction between light and molecular vibration
Raman spectrum
Laser

Ramanscattering

Wavelengthshift

via vibrational spectrum

Low-invasivenessLaser

no requirement for sample treatment

Dry and Wet condition
visible light is used for observation

Label-free

No fixation nor staining
is required!

Wavelength

White lightapplicable to difficult-to-stain sample
2850 cm-1

Wavelength tunability
405 nm532 nm1000 cm-1

780 nm2930 cm-1

1650 cm-1

Intensity/ResolutionHighLow
Invasiveness
HighLow
Resonant Raman scattering
Ramane.g) PC12 cells

Home-built multimodal microscope

What can do?

• Raman spectroscopic imaging

Label-free analysis of molecular distribution and molecular
structure
Measurement with a spectrometer (reflection arrangement)
Imaging duration: 5–30 min/image, 1–10 s/point• Second-harmonic generation (SHG) imaging

Collagen orientation and maturity, the crystallinity ofmolecules, etc.
Measurement with spectrometer (reflection configuration) or
PMT (transmission configuration)
Imaging duration: 10 s–1 minConcept:

Multifunctional spectroscopic microscopethat can be "modified" to meet your needs

Fundamental specifications
Microscope: Inverted microscope-based systemwith laser-scan / stage-scan system
LED light (UV, B, G), Phase contrast
Ex. Light:
CW 532 nm (473, 633, 785 nm will be available in the future)
Pulsed femtosecond laser 785 nm
Functions: Spectrometer (Reflection configuration)
Photomultiplier tube (Transmission configuration)
Spot detection / Spot scanning imaging
Resolution: ca. 500 nm (XY), 1,000 nm (Z)Other:Can be modified as request• Fluorescence imaging with LED light sourceFluorescence observation of stained tissue/cells
Measurement with sCMOS camera (reflection configuration)
Light source: UV, B, G excitation
Imaging duration: 0.1–10 s• Laser scanning fluorescence spectroscopic
imaging

Fluorescence observation of stained tissue/cells
Measurement with a spectrometer (reflection configuration)
Imaging duration: 1–10 s• Multiphoton fluorescence spectroscopic imaging

Fluorescence observation of stained tissues/cells in the deeppart of the living body, etc.
Measurement with a spectrometer (reflectance configuration)
Imaging time: 10 s–1 minWhat Raman microscopes can doLabel-free imaging of functional molecules and tissues
e.g. myocardial infarction diagnosis by Raman spectroscopy
Acute response

750 cm-1

Redox state ofcytochrome c

Human heart tissue
Normal

S. Ohira, T. Minamikawa et al.,Sci. Rep. (2017).

NormalInfarct

Process of MI

Old MINecrosis

T. Yamamoto, T. Minamikawa et al.,Sci. Rep. (2018)

N. Muranishi, T. Minamikawa et al.,Anal. Chem, (2014)．JP Pat. 6103700
Norm.
Nec.
Day 2
Day 5

Necrosis

Gran.
Fib.

Granulation
NormalRaman shift / cm-1

Granulation
FibrosisMolecular structural analysis of difficult-to-stain moleculese.g. Lipid Molecular Structure Analysis
Species/structurese.g. NASH diagnosis

Conformation

Lipid species(e.g. cholesterol/TAG)

DOPC

C-CStructures(Unsaturation)

C=CDPPC
Trans Gauche

Molecular Spectral Analysis
T. Minamikawa, et al. Sci. Rep, (2020)