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MB Sample ID: SA242030
| Local Sample ID: | CX_L-ON_Naïve_B3 |
| Subject ID: | SU002503 |
| Subject Type: | Other organism |
| Subject Species: | Xenopus laevis |
| Taxonomy ID: | 8355 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002503 |
| Subject Type: | Other organism |
| Subject Species: | Xenopus laevis |
| Taxonomy ID: | 8355 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| CX_L-ON_Naïve_B3 | SA242030 | FL030332 | Naïve | Factor |
Collection:
| Collection ID: | CO002496 |
| Collection Summary: | Optic nerves were collected from frogs at 7, 12, 18 and 27 weeks post optic nerve crush and subjected to lipid profiling. |
| Sample Type: | Eye tissue |
Treatment:
| Treatment ID: | TR002515 |
| Treatment Summary: | Optic nerves from each of 10 post-metamorphic transgenic Tg(Islet2b:EGFP-RPL10a) Xenopus laevis frogs, 3.5 - 5.0 cm in length, were either left untreated (naïve) or underwent a monocular surgery (operated; Fig. 1). Operated individuals were anesthetized with 0.05% ethyl 3-aminobenzoate methanesulfonate (Sigma, USA) and received either a sham surgery (sham) or a crush injury (crush) to the right optic nerve, and no treatment the left optic nerve (control; Fig. 1). |
Sample Preparation:
| Sampleprep ID: | SP002509 |
| Sampleprep Summary: | Lipids were extracted from the optic nerve tissue with a Bligh and Dyer method. The organic phase containing the lipids was removed after centrifugation and dried down with a vacuum centrifuge. The lipids were flushed with argon gas to prevent oxidation and stored at -80°C prior to analysis. Dried lipid samples were reconstitued in 49µl of isopropanol:acetonitrile 1:1 (v/v) and 1µl of EquiSPLASH™ LIPIDOMIX® Quantitative Internal Standard (330731) and sonicated for 15 minutes for total solubilization. Samples were split into two separate vials containing 25µl each, one for positive mode and one for negative mode. Reversed phase chromatographic separation was performed on Vanquish Horizon UHPLC system (Thermo) using an Accucore Vanuqish C18+ UHPLC Column. An injection volume of 5µl was used and the flow rate was 260 µl/min. Mobile phase A was 50% acetonitrile, 50% water, 5mM ammonium formate, and 0.1% formic acid. Mobile phase B was 88% isopropanol, 10% acetonitrile, 2% water, 5mM ammonium formate and 0.1% formic acid. |
Combined analysis:
| Analysis ID | AN003935 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Thermo Vanquish |
| Column | Thermo Accucore C18 (150 x 2.1mm,2.6um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap |
| Ion Mode | UNSPECIFIED |
| Units | µg/ml |
Chromatography:
| Chromatography ID: | CH002914 |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Thermo Accucore C18 (150 x 2.1mm,2.6um) |
| Column Temperature: | 55 |
| Flow Gradient: | The gradient began at 10% B for 1 min, then shifted to 30% B for 1.5min, 50% for 3.5min, 60% for 10min, 80% for 2 min, 95% for 2 min, then stayed at 100% B for 6 min before ramping down to 10% B for 2 min. |
| Flow Rate: | 260 ul/min |
| Solvent A: | 50% acetonitrile/50% water; 0.1% formic acid; 5mM ammonium formate |
| Solvent B: | 88% isopropanol/10% acetonitrile/2% water; 0.1% formic acid; 5mM ammonium formate |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003673 |
| Analysis ID: | AN003935 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The raw scans were analysed and quantified with LipidSearch 5.0 and the statistical analysis was conducted through Metaboanalyst 5.0. |
| Ion Mode: | UNSPECIFIED |
