Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA211950

Local Sample ID:	fed_fasted
Subject ID:	SU002305
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090

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Subject:

Subject ID:	SU002305
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
fed_fasted	SA211950	FL025854	fed_fasted	Treatment

Collection:

Collection ID:	CO002298
Collection Summary:	C57BL/6J (000664) and BALB/cJ (000651) mice were obtained from The Jackson Laboratory. Mice were housed at 20-22°C on a 12 h light/dark cycle with ad libitum access to food (PicoLab Rodent Diet 5053) and water. All animal studies were performed in accordance with Haigis lab protocols approved by the Standing Committee on Animals, the Institutional Animal Care and Use Committee at Harvard Medical School. For heat inactivation studies, 3 mice were used (C57BL/6J, female, 7 weeks old) and kidneys, brain halves, and liver lobes from the same individual animal were subjected to the different heat inactivation treatments (overview in Supplementary Fig. 1A, E). For desiccation experiments, 2 mice were used (C57BL/6J, male, 7 weeks old). For fasting experiments, two independent cohorts of 5 mice were used per treatment group (BALB/cJ, female, 10-11 weeks old) and mice were subjected to a 16 hour overnight fast. For HFD experiments, two independent cohorts of 4 mice were used per treatment group (C57BL/6J, female). Mice were assigned at 5 weeks old to the control diet (PicoLab Rodent Diet 5053) or HFD (Research Diets, Inc. #12492) and maintained on this diet for 4.5 months. The control diet is 4.07 Gross Energy Kcal/g. The HFD is 5.21 Kcal/g. for 8-10 weeks. Comparative MALDI MSI and LC-MS analyses of tissues were always performed on the same tissue specimens.
Sample Type:	Liver_Brain_Kidney

Treatment:

Treatment ID:	TR002317
Treatment Summary:	N/A

Sample Preparation:

Sampleprep ID:	SP002311
Sampleprep Summary:	Tissue preparation for MALDI MSI Frozen tissues were placed at -20 °C before sectioning in a Microm HM550 cryostat (Thermo Scientific™). Tissues were sectioned at 10 µm thickness and thaw mounted onto indium-tin-oxide (ITO)-coated slides (Bruker Daltonics) for MALDI MSI analysis with serial sections mounted onto glass slides for histological analyses. The microtome chamber and specimen holder were maintained between -15 °C and -20 °C. Slides were stored at -80 °C until further processing. For desiccation experiments, slides were subjected to desiccation in a tabletop vacuum desiccator before freezing. Matrix deposition A 1,5-Diaminonaphthalene(DAN)-HCl matrix solution was used for all experiments. To generate the hydrochloride derivative of 1,5-DAN, 39.5 mg of 1,5-DAN was dissolved in 500 µL of 1 mol/L hydrochloride solution with 4 mL HPLC-grade water. The solution was sonicated for 20 minutes to dissolve 1,5-DAN, after which 4.5 mL ethanol was added to yield the matrix solution. Matrices were deposited on slides and tissues using a TM-sprayer (HTX imaging, Carrboro, NC). DAN-HCl matrix spray conditions used where: a flow rate of 0.09 mL/min, spray nozzle temperature of 75 °C, and spray nozzle velocity of 1200 mm/min. A four-pass cycle was used with 2 mm track spacing and the nitrogen gas pressure was maintained at 10 psi.

Sampleprep ID:

SP002311

Sampleprep Summary:

Tissue preparation for MALDI MSI Frozen tissues were placed at -20 °C before sectioning in a Microm HM550 cryostat (Thermo Scientific™). Tissues were sectioned at 10 µm thickness and thaw mounted onto indium-tin-oxide (ITO)-coated slides (Bruker Daltonics) for MALDI MSI analysis with serial sections mounted onto glass slides for histological analyses. The microtome chamber and specimen holder were maintained between -15 °C and -20 °C. Slides were stored at -80 °C until further processing. For desiccation experiments, slides were subjected to desiccation in a tabletop vacuum desiccator before freezing. Matrix deposition A 1,5-Diaminonaphthalene(DAN)-HCl matrix solution was used for all experiments. To generate the hydrochloride derivative of 1,5-DAN, 39.5 mg of 1,5-DAN was dissolved in 500 µL of 1 mol/L hydrochloride solution with 4 mL HPLC-grade water. The solution was sonicated for 20 minutes to dissolve 1,5-DAN, after which 4.5 mL ethanol was added to yield the matrix solution. Matrices were deposited on slides and tissues using a TM-sprayer (HTX imaging, Carrboro, NC). DAN-HCl matrix spray conditions used where: a flow rate of 0.09 mL/min, spray nozzle temperature of 75 °C, and spray nozzle velocity of 1200 mm/min. A four-pass cycle was used with 2 mm track spacing and the nitrogen gas pressure was maintained at 10 psi.

Combined analysis:

Analysis ID	AN003628
Analysis type	MS
Chromatography type	None (Direct infusion)
Chromatography system	timsTOF fleX
Column	none
MS Type	MALDI
MS instrument type	QTOF
MS instrument name	Bruker timsTOF fleX
Ion Mode	NEGATIVE
Units	Da

Chromatography:

Chromatography ID:	CH002683
Instrument Name:	timsTOF fleX
Column Name:	none
Chromatography Type:	None (Direct infusion)

MS:

MS ID:	MS003379
Analysis ID:	AN003628
Instrument Name:	Bruker timsTOF fleX
Instrument Type:	QTOF
MS Type:	MALDI
MS Comments:	SCilS 2022b pro
Ion Mode:	NEGATIVE