Ultra-violet and infra-red investigations on muscle

Abstract

Infra-red absorption spectroscopy of muscle has already been carried out, using the Burch reflecting microscope (Barer, Cole & Thompson 1949: Barer, Thompson & Williams unpublished). There are considerable difficulties involved in this type of work. In the first place it is rather doubtful whether such measurements will ever be possible on living muscle owing to the presence of water, which possesses intense absorption bands in some of the most useful regions of the infra-red spectrum. It may be possible to overcome this difficulty to some extent by using heavy water which has a different absorption spectrum. It is in principle possible to obtain information similar to that given by infra-red spectroscopy, even in the presence of water, by means of Raman spectroscopy, but the technical difficulties involved, particularly fight scattering by colloids, would seem to preclude this method of attack so far as muscle is concerned. Our infra-red measurements have hitherto been confined to dried material. The results indicate that there is little prospect of working with whole muscles, as even single isolated striated fibres of the frog, rabbit and crab were usually too thick. However, it was possible to obtain good spectra in the chemically important region from 3 to 14/µon exceptionally thin single fibres or on artificially compressed fibres. An attempt was made to detect dichroism by means of polarized infra-red radiation, but to our surprise none was observed throughout the 3 to 14µrange, even though the material used showed strong birefringence in the visible region. Hr Stocken and I have recently examined certain molecular models of muscle, in the fight of the work of Ambrose, Elliott & Temple (1949) on myosin, and it now appears possible that infra-red dichroism of muscle might be expected to manifest itself only under rather special conditions. We hope to put these theoretical deductions to experimental test. As regards measurements on muscle in the ultra-violet region, the position is much more promising. It is quite possible to determine the absorption spectrum of theAorIband in living single fibres. The entire spectrum from about 230 mµin the ultra-violet to over 600 mµ, in the visible can be recorded simultaneously, using the reflecting microscope. This technique can also be used with polarized ultra-violet fight, in order to detect variation of dichroism in crystals at different wave-lengths (Barer, Jope & Perutz unpublished), and I intend to apply it to the study of dichroism in muscle fibres. Another new possibility is the observation of birefringence, as well as dichroism, in the ultra-violet. I have recently carried out experiments with a view to developing a new type of ultra-violet polarizer and it should now be possible to use the reflecting microscope as an ultra-violet polarizing microscope.