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MB Sample ID: SA352339

Local Sample ID:CTL_12dpi_R_2
Subject ID:SU003345
Subject Type:Amphibian
Subject Species:Xenopus laevis
Taxonomy ID:8355

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Subject:

Subject ID:SU003345
Subject Type:Amphibian
Subject Species:Xenopus laevis
Taxonomy ID:8355

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
CTL_12dpi_R_2SA352339FL041123RetinaSample source
CTL_12dpi_R_2SA352339FL041123ControlTreatment

Collection:

Collection ID:CO003338
Collection Summary:Retinas were collected from frogs at 12 and 27 days post optic nerve crush and subjected to lipid profiling.
Sample Type:Eye tissue

Treatment:

Treatment ID:TR003354
Treatment Summary:Optic nerves from each transgenic Tg(Islet2b:EGFP-RPL10a) Xenopus laevis frogs, 3.5 - 5.0 cm in length, underwent monocular surgery of either a left optic crush injury (crush) or sham surgery (sham). The matching controls of uninjured right optic nerves were also collected (control). Operated individuals were anesthetized with 0.05% ethyl 3-aminobenzoate methanesulfonate (Sigma, USA).

Sample Preparation:

Sampleprep ID:SP003352
Sampleprep Summary:Lipids were extracted from the retinal 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 AN005289 AN005290
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Thermo Vanquish Thermo Vanquish
Column Thermo Accucore C18 (150 x 2.1mm,2.6um) Thermo Accucore C18 (150 x 2.1mm,2.6um)
MS Type ESI ESI
MS instrument type Orbitrap Orbitrap
MS instrument name Thermo Q Exactive Orbitrap Thermo Q Exactive Orbitrap
Ion Mode POSITIVE NEGATIVE
Units peak results peak area

Chromatography:

Chromatography ID:CH004000
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:MS005019
Analysis ID:AN005289
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:POSITIVE
  
MS ID:MS005020
Analysis ID:AN005290
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:NEGATIVE
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