Hyperspectral Microscopy and Raman Imaging

Advanced Microscopy for Imaging and Characterization of Nanomaterials

HORIBA Scientific and CytoViva have integrated confocal Raman imaging, hyperspectral imaging, and advanced darkfield imaging onto a single microscope platform, essential for nanomaterial studies. By combining three rapid imaging methods, the system enables easy observation, characterization, and identification of samples at the nanoscale.

This integrated microscope utilizes the proven XploRA Plus Raman microscope from HORIBA Scientific, which provides full confocal Raman and PL imaging. The system includes variable laser illumination options (532 nm, 638 nm, or 785 nm) and an integrated spectrometer with four gratings on a motorized turret to ensure optimal resolution and coverage (gratings of 600, 1,200, 1,800, 2,400 gr/mm), along with a CCD or EMCCD for enhanced sensitivity. Data acquisition and analysis are performed via the user-friendly LabSpec6 software, with access to the KnowItAll Raman library from Horiba.

When the microscope is equipped with CytoViva's Enhanced Darkfield and Hyperspectral technology along with HORIBA's Raman capabilities, it can capture hyperspectral and darkfield images of large areas in minutes. The hyperspectral images are generated with high spatial and spectral resolution.

Using XploRA, Raman spectra of specific regions of interest within the same field of view can be quickly captured. This can be done in a single shot, across a line in the field of view, or through a full Raman map. This approach allows for cross-correlation between the Raman spectra and the broadband hyperspectral signal images, ensuring data accuracy.

Technical Features

Raman Component Specifications

Laser Options

  • Integrated up to 3 internal lasers: 532 nm, 638 nm, 785 nm
  • Other wavelengths and high-power options available upon request
  • PC controlled with AutoSwitch option for Raman/white light selection

Spectral Resolution: 1.4 cm⁻¹ to 8 cm⁻¹ depending on grating, laser, and CCD selection

Spectral Range: 50 cm⁻¹ to 4000 cm⁻¹ depending on grating and laser selection

Spectrograph

  • Flat-field spectrometer for imaging, compatible with large CCD detectors
  • High throughput with 4-position grating turret

Confocal Resolution

  • Fully confocal, adjustable confocal aperture, 500 nm XY resolution
  • SWIFT™ mode option for 10x faster Raman imaging

Detector

  • OE. TE deep-air cooled scientific CCD (-60°C), 1024 x 256 pixels
  • 16-bit, up to 1.48 MHz readout speed

Vertical Research Microscope

  • Standard 2-position illuminator with transmission/reflection illumination
  • USB image camera for image acquisition
  • Included objectives: 10x and 100x

Optical Imaging

  • Standard epi-illumination
  • Advanced darkfield illumination for transmission

Objective Sampling Optics

  • Long working distance options: 50x and 100x
  • Macro cuvette cell sampler
  • Fibre-optic probes

Software

  • OneClick functionality as standard
  • LabSpec Spectroscopy Suite
  • Available options: database, chemometrics, imaging, ParticleFinder

Weight: 35 kg

Operating Temperature: 15 - 28°C (optimal 22°C ± 1°C)

Voltage: 110/240 VAC, mains supply

Dimensions (WxDxH): 479 x 352 x 661 mm

Note: No water cooling or LN2 supply required

 

Hyperspectral Component Specifications

Spatial Resolution: 100 nm
Spectral Resolution: 2 nm
Exposure Time: 5 µs – 60 sec
Light Source: Quartz halogen with aluminum reflector
Wavelengths: VNIR (400 nm – 1,000 nm)
Maximum Spatial Scan Width: 896 µm with 10x objective
Pixel Size: 6.45 µm x 6.45 µm

Motorized Stage:

  • Travel Range: 114 mm x 75 mm
  • Motion Resolution: 10 nm

Analysis Software: ENVI 4.8
Spectral Image Display: Real-time display of the RGB image recreated from spectral data

 

CytoViva Enhanced Darkfield Optical Illumination System Specifications

System Performance:

  • Noble metals & metal oxide NPs detection: ~10 nm
  • SoftNPs (liposomes, polymers, etc.) detection: ~75 nm

Illumination System:

  • Field of View: ~400 μm
  • Numerical Aperture: 1.2–1.4 N.A.

Light Source:

  • Standard Light Sources: Solarc, halogen, mercury vapor
  • Power Supply: 100-240V AC, 50/60 Hz, 1.6A max
  • Light Guide Type: Liquid (5 mm OD, 3 mm ID)

Dimensions:

  • Maximum Height: 98 mm
  • Minimum Mounting Clearance: 51 mm
  • Weight: 0.43 kg

Requirements:

  • Immersion Oil Type A (nd > 1.515), Cargille Brand
  • Cover Slip Thickness: 0.17 mm
  • Glass Slide: 25 mm x 75 mm x 1 mm

Microscope Compatibility:

  • Upright Research Microscopes:
    • C- and O-shaped, 3-point condenser mounts
    • (Diameter and clearance requirements for CytoViva optical illumination system)
  • Inverted Research Microscopes: Requires optional inverted adapter

Objective Compatibility:

  • Low Range Objective: 10X Air, 20X Air
  • Mid Range Objective: 40X Air (N.A. = 0.7, optional iris), 40X Long Working Distance
  • High Range Objective: 60X and 100X Oil with Iris (N.A. ~1.4)