Biologic Crystals and Particles Produced in Tissue Culture. Ii. Enzymatic Responses and Environmental Transformations.

biologic crystals and particles (BCP) in this report were shown in the first paper of this series (13) as large (up to 300 z in length) RNA-staining objects (pyronin response; controlled by RNase) which appeared in cul tures of chick embryo tissues viewed by phase-contrast microscopy. Their pleomorphic shapes were described as either semirigid tubules, right-handed helices (12), ribbons, rhomboid plates, hexagonal plates, or elongated (straight or branched) filaments often arranged in whorls. These unusual objects, observed only in a special culture environment, were closely associated with the embryo chick explants. The special environment was produced in multipurpose culture chambers (2, 4†" 7,13†" 16)by lay lag broad sheets of dialysis cellophane (8†" 11,15) across the culture-containing coverslip surface. The chamber was thus separated into the following two compartments: (a) a shallow explant area (less than 50 js in depth) be tween the closely apposing cellophane and culture cover slip, and (b) a thick fluid nutrient vault area (3 mm.) between the cellophane and closing coverslip. The purpose of this second report on BCP is to : (a) identify them with proteins as shown by specific enzy matic digestion and (b) ifiustrate by phase-contrast micro graphs some of their dynamics as observed in the native environment. Their slowly changing contours, attitudes, and sizes were appreciated only when they were photo graphed by time-lapse technics which extended over a number of days, weeks, and months. The transformation of some types of tubules to right-handed helices was of particular interest. At the time of the first report, it was not definitely known that the rhomboid and hexagonal crystalline plates developed from tubules: (a) the rhom boids were formed by several types of tubular and/or helical transformations and (b) the hexagons developed from or on the opening ribbons of helices. Many varia tions in the progression of stages have been observed. Some of the transformations ifiustrated denote growth and replication processes. Other shapes of the hexagonal crystals (triangles, diamonds, and trapezoids) are shown. Models of the hexagonal system were effected by manipu lating paper soda straws and are correlated with the specific helical morphology observed. MATERIALS AND METHODS Embryo chick tis&ues.†" Fertile eggs (New Hampshire X White Leghorns) were supplied by the Texas Agriculture and Mechanical College, Department of Poultry Science. Upon delivery the eggs were set in a 40°C. incubator; embryos were selected for cultivation after 11†" 18days …