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PARISS® Hyperspectral Microscopy

The PARISS Model “PHSI” operates in field scanning mode  in darkfield scatter, fluorescence, luminescence, %transmission, %reflection and absorption.
PARISS hyperspectral microscopy work flow

Hyperspectral microscopy work flow

How PARISS hyperspectral microscopy works

Hyperspectral microscopy correlates spectra presented by the field of view (FOV) with spectra in a reference  spectral library.  An unlimited FOV is acquired by placing a sample on a slide mounted on a computer controlled translation stage on a microscope. The sample is illuminated with white light for darkfield scatter, %transmission or %reflection, or with a laser for fluorescence or other excited states. To see a video that describes how the PARISS hyperspectral microscope works click here.

Each object in the field of view will present a spectrum.  If there are tens of thousands of objects, then PARISS will acquire tens of thousands of spectra many of which are common. The challenge associated with handling thousands of unique and often common spectra is handled with custom software that sorts and classifies all spectra in the FOV.

All or some classes of spectra can then be entered into a reference spectral library. Each library spectrum can be pseudo-colored and linked to a unique target “spectral object”.  The presence of one or more pseudo-colors then confirms the location and presence of a target object.

Future samples can then be scanned, and the spectra presented by all objects in the FOV will be correlated with those in the library.  Those that meet a minimum correlation coefficient will then be “painted” onto a gray scale image. Various data processing options including counting correlated objects and various mathematical functions.

To see a video that describes how the PARISS microscope works in darkfield click here.

Creating a reference spectral library

The PARISS hyperspectral microscope acquires many thousands of spectra over an unlimited field of view (FOV).  The PARISS software evaluates all spectra and sorts them into “classes.”  One class will likely be “background spectra” the remaining spectra will be target objects.

Some, or all, classes of spectra can be added into a reference spectral library.  Each library spectrum will be pseudo colored either by the software or the instrument user.

All, or selected, spectra from the FOV that correlate with a library spectrum will then be “painted” with pixel-perfect accuracy onto a grayscale image  of the FOV.

PARISS hyperspectral microscope model: PHSI includes

  • Mounting interface: “C” mount to most research upright and inverted microscopes
  • Wavelength range: 360 to 920-nm simultaneously
  • Spectral resolution: ~1-nm at 436-nm
  • PARISS imaging spectrograph (Details)
  • Scientific CCD/CMOS spectrum detector
  • Observed image scientific CCD/CMOS
  • Data processor
  • Computer operated microscope stage
  • Custom hyperspectral microscopy software
  • Includes a MIDL calibration lamp to validate wavelength accuracy
  • Optional: NIST certified light source.
  • Optional Xenon illuminator for darkfield nanoparticle characterization
  • Calibration standards: Available MIDL wavelength calibration lamp and a “SYLPH”  NIST certified radiometric light source.
  • Sold either as an accessory to an existing microscope or as a complete system, contact LightForm.
Hyperspectral microscope image of nanoparticles internalized in cells

PARISS Hyperspectral microscopy reveals nanoparticles in cells

Histogram of distribution of spectral classes

Distribution of spectral classes in a field of view

Hyperspectral microscope image of multi-fluorescent labeled tissue

Hyperspectral microscopy used to delineate multiple overlapping fluorophores in a tissue section

Reference spectral library

The PARISS hyperspectral microscope software generates a reference spectral library.

PARISS hyperspectral software

How PARISS hyperspectral microscope software works.

PARISS Videos

Darkfield hyperspectral nanoparticle characterization

Darkfield hyperspectral nanoparticle characterization 

How hyperspectral microscopy works

How PARISS hyperspectral imaging microscopy works

PARISS hyperspectral modes of operation

PARISS hyperspectral imaging microscopy modes of operation

All PARISS Hyperspectral systems are custom configured to meet the needs of an application. The above configuration is for guidance only. Specifications can and do change without notice.