Dual Mode Fluorescence Microscopy: A New Approach to Nanoparticle and Cellular Imaging

CytoViva, Inc ’s Dual Mode Fluorescence (DMF) Microscopy offers researchers a unique imaging capability which enables simultaneous observation of both fluorescent and non-fluorescent elements within a sample in real-time without the need for post-processing overlays. Designed with simplicity and versatility in mind, DMF Microscopy combines CytoViva’s Enhanced Darkfield Microscopy with fluorescence imaging to reveal critical spatial relationships and interactions at the nanoscale, all in one field of view. With CytoViva’s DMF Microscopy, researchers can achieve new insights into complex biological interactions, offering unparalleled precision for studying nanomaterials and cellular dynamics in disciplines from biomedical research to materials science. By simplifying imaging workflows and enhancing observational power, DMF Microscopy empowers researchers to focus on discovery without the constraints of traditional imaging limitations.

The power of DMF Microscopy is most obvious during research at the nano-bio interface. The example above demonstrates this perfectly. Here, using CytoViva’s Enhanced Darkfield Microscopy and DMF Microscopy, fluorescent and non-fluorescent sample structures can be clearly observed simultaneously, and in real time.?In the images above, Au nano-rods are internalized in confluent cells, which have a DAPI (blue) stained nucleus.??

Figure 1 is an enhanced darkfield image showing nanoparticles and no DAPI label.

In Figure 2, the dual mode fluorescence optical image shows the fluorescent DAPI blue nucleus, unlabeled Au nano-rods, and other non-fluorescent cell structure.

Figure 3 demonstrates DMF Microscopy in its full fluorescent mode, with only the DAPI stain observable. Unlike CytoViva's DMF Microscopy, Figure 3 would be the only image possible with traditional epi-fluorescence or confocal fluorescence systems.

CytoViva’s DMF Microscopy uses a filter wheel with multiple excitation filters placed strategically between the light source and the enhanced darkfield microscope optics with a triple-pass emission filter in the microscope turret. With just a rotation of the filter wheel, researchers can toggle between full fluorescence and full broadband white light modes or achieve a custom blend of fluorescence excitation and white light for their specific imaging needs. This seamless modulation allows users to optimize imaging conditions directly through the microscope eyepiece or optical camera, avoiding complicated software manipulation or electronic overlay processes.

A powerful aspect of DMF Microscopy is its ability to capture both fluorescently labeled and unlabeled sample portions with exceptional contrast and clarity. For instance, in nanoparticle research, researchers can explore interactions between nanoparticles (NPs) and specific cell organelles, such as mitochondria, by labeling mitochondria with Mito Tracker Red. The fluorescent red signal highlights mitochondria, allowing researchers to observe nanoparticle colocalization with this vital organelle and assess potential interactions. In Figure 4, this capability can be clearly observed.

The integration of CytoViva’s Enhanced Darkfield Microscopy and Dual Mode Fluorescence Microscopy creates a unique research tool that combines high-contrast darkfield imaging with versatile fluorescence imaging. This powerful tool opens up new avenues for exploring nanoparticle interactions and cellular dynamics, from biomedical applications to advanced materials science, making it an essential tool for researchers seeking comprehensive, real-time insights into complex biological systems.

Please contact us at [email protected] learn more about this capability and how it can allow you to optically image, and characterize both fluorescence and non-fluorescence sample structures simultaneously and in real time.?

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