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MB Sample ID: SA245392
Local Sample ID: | B652_Hypothalamus |
Subject ID: | SU002542 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | APOE-targeted replacement mice homozygous for human E3 (B6.129P2-Apoe^tm2(APOE*3)Mae N8, Taconic #1548-F) or human E4 (B6.129P2- Apoe^tm3(APOE*4)Mae N8, Taconic #1549-F) alleles and crossed to the 5x Familial Alzheimer's Disease (5XFAD) model (MMRRC #34840, B6SJL^Tg( APPSwFlLon,PSEN1*M146L*L286V)6799Vas/Mmjax)) |
Age Or Age Range: | 3 months, 12 months, 24 months |
Gender: | Female |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU002542 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | APOE-targeted replacement mice homozygous for human E3 (B6.129P2-Apoe^tm2(APOE*3)Mae N8, Taconic #1548-F) or human E4 (B6.129P2- Apoe^tm3(APOE*4)Mae N8, Taconic #1549-F) alleles and crossed to the 5x Familial Alzheimer's Disease (5XFAD) model (MMRRC #34840, B6SJL^Tg( APPSwFlLon,PSEN1*M146L*L286V)6799Vas/Mmjax)) |
Age Or Age Range: | 3 months, 12 months, 24 months |
Gender: | Female |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
B652_Hypothalamus | SA245392 | FL030748 | Hypothalamus | Brain Region |
B652_Hypothalamus | SA245392 | FL030748 | E3_young | Group |
B652_Hypothalamus | SA245392 | FL030748 | E3/E3 | APOE genotype |
B652_Hypothalamus | SA245392 | FL030748 | Wild-type | 5XFAD Genotype |
B652_Hypothalamus | SA245392 | FL030748 | 3 months | Age |
Collection:
Collection ID: | CO002535 |
Collection Summary: | Three different mice for each experimental group (18 mice in total) were anesthetized via 5.0% isoflurane before exsanguination and transcardial perfusion with ice-cold Dulbecco’s phosphate buffered saline (DPBS; Gibco # 14040133). Following perfusion, hemibrains were quickly removed, leaving behind brainstem and cerebellum. Hemibrains were immediately placed in OCT compound (Fisher HealthCare Tissue Plus O.C.T. Compound Clear 4585) and gently lowered into isopentane (Sigma-Aldrich 2- Methylbutane M32631) in a beaker surrounded by dry ice (isopentane chilled to approximately -70°C). Hemibrains were submerged for 60 seconds, placed on dry ice, wrapped in aluminum foil, and stored at -80°C until sectioning. |
Sample Type: | Brain |
Treatment:
Treatment ID: | TR002554 |
Treatment Summary: | All mice were group housed in sterile micro-isolator cages (Lab Products, Maywood, NJ), and fed autoclaved food and acidified water ad libitum. No additional treatment was applied, with the only comparisons being across different genotypes and ages as specified in the study design table. |
Sample Preparation:
Sampleprep ID: | SP002548 |
Sampleprep Summary: | Prepared brain hemispheres were cryosectioned to 10 μm thick coronal sections at approximately Bregma -2.00 mm. Brain sections were mounted on glass slides and prepared for MALDI MSI as follows: After desiccation for one hour, slides were sprayed with 14 passes of 7mg/mL N-(1-Naphthyl) ethylenediamine dihydrochloride (NEDC) matrix (Sigma) in 70% methanol (HPLC-grade, Sigma) which was applied at 0.06mL/min with a 3mm offset and a velocity of 1200mm/min at 30°C and 10psi using the M5 Sprayer with a heated tray of 50°C. Slides were used immediately or stored in a desiccator until analysis. |
Combined analysis:
Analysis ID | AN004002 |
---|---|
Analysis type | MS |
Chromatography type | None (Direct infusion) |
Chromatography system | none |
Column | none |
MS Type | MALDI |
MS instrument type | QTOF |
MS instrument name | Waters Synapt G2 XS |
Ion Mode | NEGATIVE |
Units | Intensity per pixel |
Chromatography:
Chromatography ID: | CH002955 |
Chromatography Summary: | Not applicable (MALDI MSI) |
Instrument Name: | none |
Column Name: | none |
Column Temperature: | N/A |
Flow Gradient: | N/A |
Flow Rate: | N/A |
Solvent A: | N/A |
Solvent B: | N/A |
Chromatography Type: | None (Direct infusion) |
MS:
MS ID: | MS003750 |
Analysis ID: | AN004002 |
Instrument Name: | Waters Synapt G2 XS |
Instrument Type: | QTOF |
MS Type: | MALDI |
MS Comments: | For the detection of lipids, a Waters SynaptG2-Xs high-definition mass spectrometer equipped with traveling wave ion mobility was employed with the following parameters. The laser was operating at 2000 Hz with an energy of 300 AU and spot size of 50 μm at X and Y coordinates of 100μm with mass range set at 50 – 1000 m/z in negative mode. MALDI-MSI data files were processed to adjust for mass drift during the MALDI scan and to enhance image quality and improve signal-tonoise ratio using an algorithm available within the High-Definition Imaging (HDI) software (Waters Corp). To adjust for mass drift during the MALDI scan, raw files were processed using a carefully curated list of 20 MALDI NEDC matrix peaks (m/z), 26 small molecule MALDI peaks(m/z), and 24 lipid peaks(m/z). Files were processed at a sample duration of 10 sec at a frequency rate of 0.5 min, and an m/z window of 0.1 Da, using an internal lock mass of previously defined metabolite of taurine 124.007 m/z with a tolerance of 1amu and a minimum signal intensity of 100,000 counts. Data acquisition spectrums were uploaded to the HDI software for the generation of lipid images. Regions of interest (ROIs) were user defined by a blinded investigator using anatomical reference points based on the mouse Allen Brain atlas. For all pixels defined within a ROI, peak intensities were averaged and normalized by total ion current (TIC) and number of pixels. |
Ion Mode: | NEGATIVE |