Nanotechnology is a rapidly growing field of research and development. The growth has genrated the need for more powerful tools and equipment, especially in imaging and image analysis.

Specifically, researchers need to be able to image, characterize and map target nanostructures in their native state quickly and easily, without stains and related sample preparation.

CytoViva has tackled this need by integrating Specim’s world-class spectrographs with their patented Enhanced Darkfield (EDF) microscopy optics and customized hyperspectral image analysis software.

How hyperspectral imaging works at the nanoscale?

The images below demonstrate the power of Specim’s high spectral resolution V10E visible near-infrared (VNIR) spectrograph combined with CytoViva’s EDF microscope optics. They feature unstained mammalian cells exposed to gold nanoparticles (AuNPs) functionalized with a targeting protein. Gold nanoparticles are small gold particles with a diameter of 1 to 100 nanometers.

Figure 1 is a hyperspectral image of AuNPs in Cells with each nanoscale image pixel containing the VNIR spectral response. The image is collected with CytoViva’s EDF illuminator mounted on an Olympus BX-43 microscope frame using a 60X oil objective.

The cells are imaged in a line-scan fashion using Specim hyperspectral camera and CytoViva’s proprietary data acquisition software. An automated microscope stage moves the sample image across the slit of a Specim V10E Spectroscope integrated with a Specim sCMOS camera, creating a hyperspectral data cube.

Figure 2 is a zoom image of one of the cells in the upper right-hand corner. These images represent the power of CytoViva’s EDF microscope illumination technique as they produce a very high signal-to-noise image of nanoscale entities embedded in cells.

Figure 1. Hyperspectral image of AuNPs in Cells
Figure 2. Zoomed Image of AuNPs in Cells

Figure 3 illustrates the spectral data which can be collected and analyzed using the system. The white curve represents the cells’ and the red curve the functionalized nanoparticles’ unique spectral fingerprints.

The spectral fingerprints enable the mapping of the nanoparticles in the sample (See Figure 4). The spectral response of the cells can be further used to filter the mapping input data to prevent false positives.

Figure 3 Example Spectrum of Cells (white) and AuNPS (red)
Figure 4: Mapping of AuNPS (in red) in Cells

Researchers worldwide rely on hyperspectral microscopy

Hundreds of leading research labs worldwide use CytoViva’s hyperspectral microscopy system combined with Specim’s spectrographs and hyperspectral camera to observe nanomaterials from 1 to 100 nm interacting with cells, tissue, and other matrixes. Data generated with CytoViva hyperspectral microscopy system have been published in over 1,600 scientific journals.

CytoViva’s systems support applications such as nano drug delivery, nano bio-sensor development, and nanomaterials synthesis and characterization. Nanomaterials that can be observed include metal, metal oxide, CNTs, polymeric, and lipid-based materials. CytoViva’s hyperspectral microscopy is also developed for new clinical applications and diagnostics, such as liquid biopsies to look for cancer cells.