Scanning Confocal Near-Infrared (NIR) Microscopy "SCoNIM": A New Microscopy Technique for Three-Dimensional Histopathology

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Scanning Confocal Near-Infrared Microscopy: A New Microscopy Technique for Three-Dimensional Histopathology M.S.N.Murthy, M.G.Jones, J.Kulka, J.D.Davies, M.Halliwell, D.R.Bull, P.N.T.Wells Engineering Science and Education ( ESEJ ) published by the IEE: Invited Paper Vol. 4, (5), pages 223-230, OCTOBER 1995

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Conventional light microscopy has been the primary method of examining microstructures since the last century when Ernst Abbe demonstrated the requirements to design an optical system using light in the visible wavelengths. To image microstructures within tissue in detail it is necessary to process the tissue and obtain thin ( 5 m) physical sections by using destructive physical sectioning techniques.
This work describes a microscopy technique using near-infrared light (700 nm-1500 nm) as the source. This has a facility to image thin sections of thick tissue specimens non-destructively. Images of contiguous two dimensional thin sections obtained serially can be combined to generate a three dimensional image of the structures being imaged. The prototype infrared confocal microscope developed in Bristol is one part of the projected composite acoustic-infrared-visible confocal microscope system on the same base: to enable visualisation and comparison of the images of the same tissue obtained by these three different modalities. The results obtained with the NIR system have shown the functional capability of scanning confocal NIR microscope. The limitations of the system are being tackled now with a view towards obtaining as good a resolution and scanning speed as is possible. Such a system with better axial and lateral resolution capability in the transmission and reflection mode is currently under development. The utilisation of the two modalities of imaging with the same microscope i.e. in the transmission and reflection modes can provide two different sets of information for analysis and diagnosis. Hopefully it will help provide further identifiable features which may not have been obvious earlier with images obtained from conventional visible optical microscopes.
Some of the images obtained from breast tissue can be seen by clicking on the hypertext annotations:
Ducts with micro-calcification clusters: Conventional microscope image and three SCoNIM images
SCoNIM images compared with an image obtained using an Infrared Microscope