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Chinese Hamster Chromosomes

This RCIM photomicrograph is of Chinese Hamster Chromosomes, anaploid (or incomplete diploid chromosome complement). The chromosomes were routinely prepared, Giemsa stained, and photographed using a Zeiss Photoscope III (Carl Zeiss, Inc., Thornwood, N.Y.) Ektachrome ASA 400 film with a 23 to 30 second exposure. An internegative was made for enlargement. By R.L. Peer, 1998, Bristol, Myers, Squibb.

For this type of RCIM the setup was as follows:
A 250 W CSI lamp in an Opti-Quip (Opti-Quip, Inc., Highland Mills, N.Y.) lamp housing, was set up about 24” from the microscope. The illumination passed through a heat absorption and heat reflection filter (Carl Zeiss, Inc., Thornwood, N.Y.) and then directly into the Fiberoptic Light Scrambler.

The Output Module of the Light Scrambler was attached to the epi port of the Photomicroscope III and the light passed through a plano-concave prism, a polarizer, a 1mm diameter pinhole, a 546nm +/-3nm narrow band filter, analyzer and thence through a 63X Zeiss Antiflex objective (top lens had a _ wave retardation plate). The light was set with the Antiflex objective and controlled with a polarizer/analyzer combo. The polarizer/analyzer were fabricated of quartz, coated for general 400-700nm use by Vino Vats (Karl Lambrecht Corp., Chicago, IL). The other retarders and prisms were produced and coated similarly.

Further notes by the author:
The results in the photo are not due to Giemsa fluorescence. I checked that out early in my work in RICM. All alignments are extremely critical with RCIM, especially the Light Scrambler and the 1mm pinhole. Excellent optics and high quality prisms are also a must.

References:

• von de Hulst, H.C. Light Scattering by Small Particles, 1981 (1957) Dover
• Ditchburn, R.W., Light 1991 (1961) Dover
• Born & Wolf Principles of Optics, 6th edition, 1980 (Pub?)
• Hecht, Eugene, Optics, 2nd edition, 1987, Addison-Wesley (also his Optics in Schaum’s Outline Series)
• Buchwald, I. Z., 1989 Rise of the Wave Theory of Light. Univ. of Chicago Press
• Sommerfeld, Arnold Optics, Academic Press, Papers for Small Holes
• Bethe, Hans, 1944. Theory of Diffraction by Small Holes. Physical Review., 66:163
• Bouwkamp, C.J. 1950. On Bethe’s Theory of Diffraction by Small Holes, Philips Res Reports. 5:401