Nanotechnology & Surface Metrology

Cytoviva – Microscope System


Co-developed by Auburn University and Aetos Technologies, Inc., CytoViva’s optical microscopy technology is a 2006 and 2007 recipient of the prestigious R&D 100 award. This award is granted annually to the market’s most innovative new technologies. In 2007, CytoViva also received a Nano50TM award, for its contribution to the fast growing world of nanotechnology research.  In 2009, US patents No. 7,542,203 and 7,564,623 were issued for CytoViva’s advanced microscopy illumination optics.


CytoViva Enhanced Darkfield Imaging

Cytoviva Microscope System – Enhanced Darkfield Imaging

Specifically designed to support research in nanotechnology and infectious disease, CytoViva employs a patented (US patents No. 7,542,203, 7,564,623) darkfield-based optical illumination system. This structured illumination technology replaces the standard condenser on a research grade microscope. By improving the alignment and focus of darkfield or oblique angle illumination, the technology enhances signal-to-noise of nanoscale samples up to seven times over standard darkfield optics. This enables scientists to optically observe a wide range of nanoscale materials quickly and easily in solution, live cells, tissue and materials based matrices. In addition, non-fluorescent live cells and pathogens can be easily observed at a level of detail not possible with traditional optical imaging techniques such as phase contrast or differential interference contrast.

When using the CytoViva Dual Mode Fluorescence system, researchers can also observe the interactions between fluorescently labeled nano-particles or bacteria and live unlabeled cells. This unique capability can eliminate the need to create computer enhanced overlay images which require two different illumination methods and advanced software programs. Finally, when combined with CytoViva’s Hyperspectral Imaging capability this high signal-to-noise microscopy method enables researchers to spectrally characterize and map nanoscale samples in a wide range of environments.

  • Sub 100nm resolution of live cells and nano-materials
  • Fluorescent and non-fluorescent sample structure simultaneously

Sample Image

Cytoviva sample Image

CytoViva Video AuNPs in live cell culture

This video illustrates the interactions of AuNPs in a live epithelial cell culture. The video was captured using an optical microscope equipped with the patented CytoViva high signal to noise illumination system.

The System:

CytoViva Enhanced Darkfield Imaging

High Resolution Illuminator


The CytoViva high resolution illuminator replaces the standard microscope condenser. The specialized illuminator focuses fixed-geometry, highly collimated light at oblique angles on the sample. This serves to dramatically improve contrast and signal-to-noise ratio which allows for optimized resolving power and detection capability of non-fluorescing samples.

See video:

Cytoviva High Resolution Illuminator Specification

CytoViva Enhanced Darkfield Imaging

Dual Mode Fluorescence (DMF)

The Dual Mode Fluorescence (DMF) module allows for the observation of both fluorescent and non-fluorescent sample portions simultaneously and in real-time. Samples are viewed directly through the microscope eyepiece and captured using a standard microscope camera without the need for complicated software or electronic manipulation.

Cytoviva Dual Mode Fluorescence Specification

CytoViva Enhanced Darkfield Imaging

Custom Environmental Chamber

The CytoViva environmental chamber supports high resolution, long-term studies of live cells, while enabling real-time, simultaneous observation of fluorescent and non-fluorescent sample portions.

Manufactured by Warner Instruments, the CytoViva environmental chamber is a modified version of the Warner RC-30 Confocal chamber. This modification supports oil immersion contact with the CytoViva high resolution illumination system as well as an oil immersion microscope objective.

The CytoViva system supports all traditional environmental chamber applications including perfusion, temperature control and gasses. Operating as a closed bath system, the CytoViva compatible chamber works with both inverted and upright research grade microscopes.

Now researchers can observe, track and image the interactions between live cells and other elements such as fluorescently labeled nano-particles and bacteria. These observations can be made over hours or even days.

Traditional cell biology and neuroscience applications are also supported by this system.
With the CytoViva system and environmental chamber you can observe fluorescently labeled cellular components simultaneously with the unlabeled portions of the cell.

The CytoViva Environmental Chamber can also be utilized as a fully functional micro-fluidics platform supporting applications in drug delivery, tissue engineering and layer assembly of nano-materials.

Microscope Compatibility

The CytoViva system is available as a comprehensive optical microscope solution. This can include an Olympus research grade microscope, camera, light source and image analysis software.

In many instances however, researchers will add the CytoViva capability to their existing microscope system. The CytoViva high resolution illuminator is designed to fit onto the condenser mount of most upright research grade microscopes. It is the only CytoViva component that attaches directly to the microscope. Additionally, the CytoViva system can be adapted for operation with most inverted microscopes.

Camera Compatibility

CytoViva ® provides a wide range of research grade microscope cameras available from a variety of vendors including Q Imaging, Dage MTI and Optronics.

Proper camera selection will be based on your specific application and needs. For example, capturing a simultaneous, real-time image of both fluorescence and non-fluorescent sample structure will typically require a digital color camera. However, maximizing the resolution of certain images may require a cooled monochrome camera solution.

At CytoViva we can assist you in determining the most appropriate image capture capability for your application and provide this as part of an overall customized system solution.

Objective Compatibility

To maximize the benefits of CytoViva, your microscope must be equipped with the proper objectives. Most applications will require that you have the objectives within the ranges listed below. We encourage you to contact us to determine your specific needs.

Mid : 40x oil, air or water NA < 1.0 iris optional

High : 60x – 100x oil immersion NA ~1.4 w/iris

Image Analysis Software

CytoViva is unique in its ability to create high contrast images while observing both fluorescent and non-fluorescent sample structure simultaneously and in real time. This enables researchers to capture images never before possible with an optical microscope. The ability to gather quantifiable data from these real time images can be critical.

As part of an overall system solution, CytoViva provides an image analysis capability that can enable you to determine variables such as fluorescence intensity, counts and distance measurements. This capability may be included as part of a camera control operating system or provided on a standalone basis based on your needs. To learn more about our image analysis capability, click on the link below to contact a technical specialist.

CytoViva ® Optical Performance

CytoViva allows one to achieve optimized resolution and detection over traditional optical imaging techniques with living, fixed or non-biological samples. These capabilities result primarily from the exceptional contrast (high signal to noise ratio), which allows utilization of non-diffraction limited phenomena and improved point spread function.

This improved signal to noise ratio performance is achieved through:

The precise optical alignment in the fixed geometry of the unit

Ultra-efficient management of light within the optical path of the unit

Producing both Koehler illumination and main features of critical illumination

The resolving power of a microscope depends on the properties of the objective and the degree of coherence of the light incident upon the object. With traditional Critical illumination, a uniformly bright light source is placed close behind a diaphragm and is imaged by a condenser on to an object plane of a microscope objective. The light source is focused on the object. With traditional Koehler illumination, a converging (collimating) lens is placed close to the diaphragm and forms an image of the source light in the focal plane of the condenser, which includes a condenser diaphragm. Rays from each light source point then emerge from the condenser as a parallel beam. The light source is focused on an aperture of the condenser.

With CytoViva, complete Koehler illumination and a main feature of Critical illumination are achieved using a novel illumination system. Koehler illumination is pre-aligned in the device by fixing the light source precisely on the entrance slit of the condenser. This allows the user to adjust a focus point on the sample, which is a useful feature of Critical illumination, and is achieved when the condenser is aligned with the objective to find the focal point on the sample. Thus, Koehler illumination is initially fixed, and then CytoViva can be adjusted (up or down) to find the proper position and size of an illuminated spot for Critical illumination. Both Koehler and Critical illumination are not generally achievable in traditional microscopy. The resulting high signal to noise ratio of CytoViva allows the user to observe previously obscured non-diffraction limited optical effects.

The resolution of conventional optical microscopy is limited by the wave nature of light and is conventionally defined by the Rayleigh criterion; this is commonly referred to as diffraction limited resolution (limit of ~240 nm). With CytoViva, non-diffraction limited optical effects and an improved point spread function are produced that extend its resolution well beyond diffraction limits.*

*90nm (lambda/5) resolution in optical transmission microscope with annular condenser: Optics Letters, Vol. 31, Issue 19, pp. 2855-2857 Arnold Vainrub, Oleg Pustovyy, and Vitaly Vodyanoy.

What Scientists Are Saying

J. Paul Robinson, PhD, Director of Purdue University’s Cytometry Laboratory, Professor of Basic Medical Sciences and Biomedical Engineering, Purdue University comments…
“”It’s like not knowing you need glasses. You don’t know what you can’t see. Then someone hands you a pair-and the world is clear with amazing detail.””

John A. Smith, MD, PhD, MMM, Divisional Director, Department of Pathology, University of Alabama-Birmingham says it well …
“”Looking through CytoViva, you are face to face with living biology. You see into the world of cell biology that you didn’t know existed. You are visualizing the future of underlying biological processes as it merges with the present.””

Dr. Elaine Coleman, Associate Professor, Auburn University, Department of Anatomy, Physiology and Pharmacology comments…
“”Its capabilities in cell culture research are astounding. I have purchased a unit for my future research because its capabilities are unique for observing live cell cultures.””

Resolution of 90 nm (λ/5) in an optical transmission microscope with an annular condenser

Arnold Vainrub, Oleg Pustovyy, and Vitaly Vodyanoy
(pdf symbol)  Optics Letters 31 (2006)2855-2857

Resolution of 90 nm was achieved with a research microscope simply by replacing the standard bright-field condenser with a homebuilt illumination system with a cardioid annular condenser. Diffraction gratings with 100 nm width lines as well as less than 100 nm size features of different-shaped objects were clearly visible on a calibrated microscope test slide. The resolution increase results from a known narrower diffraction pattern in coherent illumination for the annular aperture compared with the circular aperture. This explanation is supported by an excellent accord of calculated and measured diffraction patterns for a 50 nm radius disk.