For experiments in which feeding was assessed, larvae that had been continuously fed with an excess of paramecia from 5 dpf were either isolated or placed in groups of 3, in 35-mm dishes (3 ml of embryo water), in the absence of food. Paramecia was labeled using lipid dye (DiD’ solid, D-7757, Thermo Fisher Scientific, dissolved in ethanol) for ~2 hrs (5 µl of 2.5 mg/ml working solution per 1 ml of concentrated paramecia, followed by spinning down, rinsing, and reconstitution in deionized or embryo water). After 2 hrs of food deprivation, fluorescent-labeled paramecia was added followed by a quick fixation after 30 min (full protocol is described in Wee et al. (2019)43 and available here: https://bio-protocol.org/prep1116 ). Sensory cues were generated and presented as described above for pERK experiments. Fixed larvae were subsequently distributed into 96-well flat-bottom dishes and imaged using the AxioZoom V16 (Zeiss) and analyzed using Fiji software (3D Objects Counter, custom software also available on www.github.com/carolinewee ).
Did solid
Manufactured by Thermo Fisher Scientific
Sourced in United States
DiD' solid is a fluorescent dye used in various biological applications. It functions as a lipophilic tracer, capable of labeling cell membranes and lipid-rich structures. The dye exhibits excitation and emission maxima suitable for detection with standard fluorescence instrumentation.
Automatically generated - may contain errors
Lab products found in correlation
Axio zoom v16, by Zeiss (2 mentions)
Anti pdgfrα, by BioLegend (1 mentions)
2 iminothiolane hcl, by Merck Group (1 mentions)
Pen strept, by Thermo Fisher Scientific (1 mentions)
Prestoblue cell viability reagent, by Thermo Fisher Scientific (1 mentions)
Siteclick antibody azido modification kit, by Thermo Fisher Scientific (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
Fluorescamine, by Merck Group (1 mentions)
Labrafac, by Gattefosse (1 mentions)
2 protocols using did solid
1
Feeding Behavior Quantification in Larval Fish
2
Targeted Delivery of Therapeutic Nanoparticles
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Labrafac® provided by Gattefosse SA (France) and Lipoïd® S100 by Lipoid Gmbh (Germany). Kolliphor HS15®, poly-d-Lysine hydrobromide (70000-150000 Da), fluorescamine, 2-Iminothiolane•HCl and propidium iodide were purchased from Sigma (United-States). 1,2-Distearoyl-sn-glycero-3-phosphorylethanolamine-PEG (2000) -Maleimide (DSPE-PEG-Maleimide) and NH 2 -PEG (5000) -Dibenzocyclooctyne (NH 2 -PEG-DBCO) were purchased from Nanocs (United-States). NH 2 -PEG (5000) -Maleimide was purchased from Creative PEGWorks (United-States). Anti-PDGFRα was purchased from BioLegend (United-States). Anti-rat-HRP, DMEM (41966-029), HEPES, Pen/Strept, DMEM 0.05% trypsin-EDTA (1x), Presto blue Cell Viability Reagent, DiD'solid, SiteClick™ Antibody Azido Modification Kit and Alexa-488-Phalloidine were purchased from Thermo Fisher Scientific (United-States). Plateletderived growth factor A (PDGFAA) and fibroblast growth factor 2 (FGF) were purchased from Peprotech (United Kingdom). rmPDGFRA/Fc chimera (soluble PDGFRα) has been purchased from R&Dsystems. Spectra-Por Float-a-lyzer G2 300 kDa dialysis membrane and Rotilabo® syringe sterile 0.22 μm filters were purchased from Carl Roth GmbH (Germany).
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