Primo star compound microscope

Colonies started from protoplast preparations were used rather than tissue fragments produced by routine propagation techniques. Week‐old tissue was used to generate protoplasts (as above) and 100 μl of liquid plating medium (PpNH₄ containing 8.5% mannitol and 10‐mM CaCl₂) containing roughly 60,000 protoplasts was added to six wells of a 12‐well plate for each of the following lines: WT, TC, sbh‐1, sbh‐2a, and sbh‐3 (Table S3). For each group of six wells, three contained PRMB with 10 μM t17:0, and three did not. After 4 days each cellophane disk was transferred to a small petri dish with PpNH₄ medium, with or without 10 μM t17:0. One cellophane disk of each line was removed for imaging (and tissue harvesting for LCB analysis) at 8, 14, and 35 days following protoplast formation. Samples were imaged using a Zeiss Stemi dissecting microscope and a Zeiss Primostar compound microscope, both equipped with Excelis color digital 1080p cameras with displays. Chloronema and caulonema cell dimensions (length and width) were measured from calibrated images using ImageJ. Apical cells were not included in these measurements. The areas of gametophore phylloid cells were determined by tracing their outline using ImageJ. Confidence intervals for means and standard deviations of cell dimensional measurements were calculated by bootstrapping with 1000 resampled parameters.

The bats were sacrificed by an overdose of sodic pentobarbital injectable and preserved and fixed in a 10% formaldehyde neutral solution. The bats' abdominal and thoracic cavities and organs were examined with a stereoscopic microscope. The intestine was divided in three sections: 1) duodenum, 2) small intestine (jejunum and ileum), and 3) large intestine and rectum. The site of infection and the number of parasites found were registered and the specimens which were found detached from the host during the necropsy were not assigned to any particular intestinal section. Permanent and transitory slides were made in order to identify the helminths. The trematodes were dehydrated gradually in ethylic alcohol (70%, 96%), stained with Grenacher's carmin, and mounted in Canada balsam. The cestodes and nematodes were cleared with Aman's lactophenol. All parasites were photographed and measured using a Zeiss Primo Star compound microscope equipped with a digital camera (Zeiss Axiocam ERc 5s). One specimen of each species was deposited in the Colección Nacional de Parasitología, Museo Argentino de Ciencias Naturales Bernardino Rivadavia, Buenos Aires, Argentina (MACN-Pa).