In vivo, real-time confocal imaging.

Area of Science:

• Biomedical Engineering
• Microscopy
• Cell Biology

Background:

• Confocal microscopy is a powerful technique for high-resolution imaging.
• In vivo imaging allows for the study of biological processes in their natural state.
• Existing methods often require tissue fixation, limiting dynamic studies.

Purpose of the Study:

• To adapt a tandem scanning confocal microscope for real-time, non-invasive in vivo imaging of cells.
• To evaluate the system's capability in visualizing cellular structures within various living tissues.
• To establish a new paradigm for studying cellular structure and function over time.

Main Methods:

• Development and adaptation of a tandem scanning confocal microscope for in vivo applications.
• Utilized a 20x BioOptics surface contact objective for imaging.
• Employed a DAGE Mti low light level SIT camera and Gould IP9527 image processor for image acquisition and processing.
• Compared in vivo images with conventional bright-field and scanning electron microscopy of fixed tissues.

Main Results:

• Successfully obtained real-time, non-invasive cellular images from various living tissues (cornea, kidney, liver, etc.) in rabbits and rats.
• Achieved detailed cellular visualization comparable to traditional microscopy of fixed tissues.
• Demonstrated the ability to image cells up to 1 mm deep within opaque organs.

Conclusions:

• In vivo confocal imaging offers remarkable detail of living cells, comparable to fixed-tissue microscopy.
• This technique represents a fundamentally new approach for studying cellular structure and function sequentially over time.
• Further refinements promise enhanced resolution and unprecedented biological insights.