Paramecia Labeling and Fish Larvae Feeding

Paramecia were collected by centrifugation at 3,000 rmp for 5 min and quantified by optical density measurement at OD500 nm as described (45 (link)). The paramecia were diluted to a density of 1000 paramecia per ml and incubated with 4-(4-(didecylamino)styryl)-N-methylpyridinium iodide (4-Di-10-ASP) at 25 μg/ ml for 1 hr (Invitrogen, Carlsbad, CA, USA). The paramecia were then washed with water three times for 10 min each time to remove the residual 4-Di-10-ASP. Fish larvae were fed with 4-Di-10-ASP labeled or unlabeled (control) paramecia at 1000 paramecia/ml for two hours. The fish larvae were photographed immediately using a GFP2 filter on a MZ16F stereoscopic microscope (Leica Microsystems). The images were analyzed using Image J software to get the fluorescence intensity.

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Li S., Wen H, & Du S. (2019). Defective sarcomere organization and reduced larval locomotion and fish survival in slow muscle heavy chain 1 (smyhc1) mutants. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 34(1), 1378-1397.

Publication 2019

4-Di-10-ASP MZ16F stereoscopic microscope

Centrifugation Fish Fluorescence Iodide Larvae Microscope N methylpyridinium Optical Paramecia

Corresponding Organization :

Other organizations : University of Maryland, Baltimore, Ocean University of China

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Variable analysis

independent variables

Presence of 4-Di-10-ASP (labeled or unlabeled paramecia)

dependent variables

Fluorescence intensity of fish larvae

control variables

Paramecia density (1000 paramecia/ml)
Incubation time of paramecia with 4-Di-10-ASP (1 hr)
Paramecia washing steps (3 times for 10 min each)
Feeding duration of fish larvae (2 hours)

controls

Positive control: Fish larvae fed with 4-Di-10-ASP labeled paramecia
Negative control: Fish larvae fed with unlabeled (control) paramecia

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