Multiphoton and Raman microscopes

Multiphoton confocals work by using an infrared laser at double or triple the wavelength necessary to excite a fluorophore.

When two or three photons interact with the fluorophore at the same time, it is excited and emits fluorescence.  The advantage of infrared light is that it can penetrate deep in tissues, unlike visible light which scatters.  Therefore, a multiphoton can image deep in thick samples – provided the emitted signal is strong enough to be captured! It helps if the sample is cleared.

Miltenyi TRIM

The Miltenyi TRIM is a multiphoton confocal microscope equipped with two overlapping tuneable infrared lasers (680 to 1300nm) and a fixed line at 1064nm.  This microscope can be configured upright as well inverted, giving complete flexibility of sample access.  It has  a 10x, 20x, 25x and 40x lenses, as well as 11 photoreceptors.  This microscope has multiple filter sets to detect many dyes and fluorescent proteins. The TRIM is also capable of imaging the structure of the imaged tissue using two label-free modes: 1) through second harmonics,  -a process that does not require dyes but relies on autofluorescence; and 2) Raman microscopy -which determines the molecular composition of a sample based on photon-induced specific chemical bond excitation.

Special applications and features:

  1. Multiphoton imaging goes deep in sample
  2. Upright and inverted microscope
  3. Second Harmonics to detect autofluorescence in the tissue and signal context
  4. Raman/CARS microscopy for molecular identification.  Raman works specially well with fatty molecules
  5. Resonant scanner mode for fast imaging

Location: IRR Translational Imaging Hub, contact IRR.Imaging@ed.ac.uk for access.

Leica Stellaris for Coherent Raman Scattering (CRS)

A fully integrated confocal and CRS imaging platform will be coming to the IRR Imaging Hub in Q2 2026, funded by a UKRI-BBSRC grant (UKRI2249).

 Its capability will include stimulated Raman scattering (SRS), coherent anti-Stokes Raman Spectroscopy (CARS), second harmonic generation (SHG) and two photon fluorescence (TPF) as well as confocal microscopy. Designed for live imaging, the microscope will have an incubation chamber and a range of lens magnifications (10 x, 20 x, 40 x).

Location: IRR Imaging Hub, contact IRR.Imaging@ed.ac.uk for sample imaging.

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