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Ramacle® is an all-encompassing software package written for complete instrument control and data handling for all multimodal techniques on the RMS1000.
Just like the hardware, the software operating the RMS1000 adapts to your research need. Ramacle controls all RMS1000 functions with a straightforward design concept. It focuses on all modern Raman spectroscopy applications, and beyond. For example, running Raman and FLIM maps easily in one comprehensive platform.
Figure 1: Ramacle software interface
With Ramacle, you can easily navigates from microscope set up to measurement set up and offers computer-controlled measurement conditions such as laser excitation, grating, pinhole size etc. On the RMS1000, Ramacle comes with standard data acquisition methods such as single measurements, multiple and accumulated scans, kinetic scan, as well as a comprehensive suite of mapping techniques e.g. 2D, 3D, SurfMAP, and FastMAP. Upgrades and accessories such as temperature stages and multiwell plates unlock additional software features.
2D mapping and 3D mapping allow the user to see distribution of components, as well as revealing areas under high stress or strain, and defects. If using short exposure times FastMAP® is also available to reduce total acquisition times.
Figure 1: 2D Raman mapping showing nanocrystalline silicon and strain (pink, yellow, red) in a defective wafer
Figure 2: 3D Raman map of a pharmaceutical oil-in-water emulsion with TiO2 inclusions
SurfMAP® is a tool which allows accurate mapping of uneven surfaces giving the user assurance that their measurements are in focus and data is reliable. Ramacle builds a surface image which can be viewed in 2D and in 3D topography view. The software then acquires the spectral map moving the stage in X, Y, and Z for perfect laser focus.
Figure 1: Raman SurfMAP® of carbon fibres tracking the D and G bands
Figure 2: Raman SurfMap of an antipsychotic pharmaceutical drug
RaFLIM is a standout technique on the RMS1000. Combining Raman and FLIM to create maps of the same area, with the same spatial resolution (laser dependent). Operating measurement acquisition and data analysis in Ramacle for ease of use and continuity.
Figure 1: Average lifetime and fluorescence intensity image of a Convallaria Rhizome section stained with Acridine Orange. FLIM reveals variation in lifetime across lignified and pectin rich cell walls (2 exponential tail fitting).
Figure 2: Biochemical analysis of woody plant cell walls using our multi-modal micro-spectroscopy approach.
The RMS1000’s external laser port unlocks limitless potential for advanced techniques like multiphoton imaging. Techniques such as two-photon excited fluorescence (2PEF) and second harmonic generation (SHG) are ideal for studying biological samples. Both 2PEF and SHG demand extremely high excitation intensity, provided by a mode-locked femtosecond pulsed laser.
Figure 1: Mouse intestine stained with Alexa Fluor® 568
Figure 2: Photon lifetime imaging of mouse kidney sample
The sample of the mouse intestine (Figure 1) stained with Alexa Fluor® 568. Simultaneous measuring of spectral 2PEF and SHG from Alexa Fluor® 568 dye and fibrillar collagen respectively. Measurement using femtosecond laser and CCD camera. Using the same laser, two photon lifetime imaging was carried out an a sample of mouse kidney (Figure 2). This time a Hybrid Photodetector was used with TCSPC.
2D materials are also often studied with different measurement techniques, the RMS1000 can easily be configured to carry out all imaging techniques on the same sample area providing a full suite of complimentary information. Raman and PL images were collected using a 532 nm laser, whilst the SHG was acquired using a femtosecond laser.
Figure 3: Sample of WSe2 crystal. The darkfield (DF) image reveals areas of different heights. Raman, PL, and SHG images can be used correlatively to identify layer number, and highlight areas of stress, strain, dopants, and defects
The RMS1000 is built with flexibility in mind. A choice of excitation lasers (internal or external to the system) and associated laser rejection filters (both edge and notch) are available. Additionally pulsed diode lasers such as our EPL series can be added if fluorescence lifetime measurements are required.
The RMS1000 can be fitted with both standard and long focal length spectrographs. This allows the system to be configured for ultimate spectral resolution, maximum optical throughput, low wavelength and ultraviolet spectral ranges, or for highly sensitive measurements.
A choice of CCD, EMCCD and InGaAs detectors are available with a maximum of four detectors being integrated per system. For fluorescence lifetime measurements, single photon counting detectors can also be integrated.
The RMS1000 uses either an upright or inverted microscope platform which is compatible with all standard microscopy applications.
Brightfield, darkfield, polarised light, differential interference contrast (DIC) and fluorescence accessories are all available. A large choice of high-quality microscope objectives, high-performance visualisation cameras, collimators and illuminators can be added to the microscope at any time.
Manual or motorised stages are available. The motorised stage allows automated XYZ Raman and fluorescence maps to be obtained and generated through Ramacle. Additional sample stages are compatible with theRMS1000, such as temperature stages, electrochemical cells, and diamond anvil cells.
Other accessories such as a polarisation kit, cuvette holder, and a Class I laser safety enclosure are also available to further expand the capabilities, flexibility and safety of your RMS1000 system. Coupling to other fluorescence spectrometers is also possible.
