Fresh human tonsils were snap frozen in liquid nitrogen following excision as described above. Tissue sections of 10 μm thickness were prepared using an HM525 NX cryostat (Thermo Scientific, Waltham, MA) operated at −20°C, and thaw-mounted onto the conductive side of indium tin oxide (ITO)-coated slides. Slides were placed in a vacuum desiccator at −10 mmHg for at least 15 min before matrix deposition. Matrix deposition was performed with a TM-Sprayer (HTX Technologies, Chapel Hill, NC). For positive mode mass spectrometry imaging (MSI) experiments, 2,5-dihydroxybenzoic acid (DHB; Alfa Aesar, Ward Hill, MA) was sprayed onto the glass slide after tissue mounting at a concentration of 30 mg mL−1 with 12 passes/cycles (approx. density = 0.001667 mg/mm2). For negative mode MSI, 1,5-diaminonaphthalene (DAN; Sigma-Aldrich, Burlington, MA) was sprayed onto the glass slide after tissue mounting at a concentration of 5 mg mL−1 with 8 passes/cycles (approx. density = 0.009600 mg/mm2). Universal sprayer settings were as follows: nebulization gas pressure = 10 psi, track spacing = 3 mm, velocity = 1200 mm min−1, CC pattern.
1 5 diaminonaphthalene dan
Manufactured by Merck Group
Sourced in Canada, Thailand, United States
1,5-diaminonaphthalene (DAN) is a chemical compound used in various laboratory applications. It is a white to pale yellow crystalline solid. DAN serves as a reagent and intermediate in organic synthesis.
Automatically generated - may contain errors
Lab products found in correlation
Tm sprayer, by HTX Technologies (1 mentions)
2 5 dihydroxybenzoic acid dhb, by Thermo Fisher Scientific (1 mentions)
Hm525 nx, by Thermo Fisher Scientific (1 mentions)
Acetonitrile acn, by Thermo Fisher Scientific (1 mentions)
Ethanol, by Merck Group (1 mentions)
Trifluoroacetic acid tfa, by Honeywell (1 mentions)
Chloroform, by RCI Labscan (1 mentions)
Formic acid, by Honeywell (1 mentions)
Tetrahydrofuran, by Thermo Fisher Scientific (1 mentions)
Ethanol, by Thermo Fisher Scientific (1 mentions)
Methanol, by Thermo Fisher Scientific (1 mentions)
Chloroform, by Thermo Fisher Scientific (1 mentions)
2 5 dihydroxyacetophenone, by Merck Group (1 mentions)
Hplc grade acetonitrile, by Thermo Fisher Scientific (1 mentions)
Lipid standards, by Avanti Polar Lipids (1 mentions)
5 protocols using 1 5 diaminonaphthalene dan
1
Mass Spectrometry Imaging of Frozen Human Tonsils
2
Sublimation Apparatus for DAN Synthesis
3
Cryosectioning and MALDI Imaging of Brain Tissue
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Brains were attached to a specimen holder on dry ice with distilled water. Using a cryostat, 10 μm thick coronal brain sections were thaw-mounted onto indium tin oxide (ITO) coated glass slides (Hudson Surface Technology Inc., Old Tappan, NY, USA). Slides were dehydrated for 15 min in a vacuum chamber and then coated with 1,5-diaminonaphthalene (DAN, Sigma–Aldrich, Oakville, ON, Canada) matrix using a sublimation apparatus (Chemglass Life Sciences, Vineland, NJ, USA). DAN matrix was sublimated for 9 min at 130°C sand bath temperature. Following matrix application, a calibration mixture of five standard proteins (4700 Calibration Mixture, AB Sciex, Concord, ON, Canada) was mixed with alpha-cyano-4-hydroxy-cinnamic acid matrix (CHCA, Sigma–Aldrich) and 0.75 μl spots were applied on DAN matrix-free areas of the glass slide in the vicinity of the section to be imaged. Once calibration spots had dried, slides were stored at -20°C overnight. The following day, glass slides were thawed and dehydrated in a vacuum chamber for 15 min.
4
Luminescent Conjugated Oligothiophene Characterization
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All chemicals and solvents were
used without further purification: acetic acid (Cat.#: 64197, VWR
Chemicals, Radnor, PA, USA), acetonitrile (ACN, Cat.#: 75058, Fisher
Scientific, Waltham, MA, USA), chloroform (Cat.#: 67663, RCILabScan,
Bangkok, Thailand), 1,5-diaminonaphthalene (DAN, Cat.#: 56451, Sigma
Aldrich, St.Louis, MO, USA), 2′,5′-dihydroxyacetophenone
(DHA, Cat.#: D107603, Sigma Aldrich), ethanol (Cat.#: V002075; Sigma
Aldrich), formic acid (FA, Cat.#: 56302, Honeywell), and trifluoroacetic
acid (TFA, Cat.#: 40967; Honeywell, Charlotte, NC, USA). Luminescent
conjugated oligothiophene (LCO) tetramer formyl thiophene acetic acid
(q-FTAA) and heptamer formyl thiophene acetic acid (h-FTAA) were obtained
from Prof. Peter Nilsson, Department of Chemistry, Linköping
University. Water was obtained from a Synergy UV water purification
system (Milli-Q, Merck Millipore, Darmstadt, Germany).
used without further purification: acetic acid (Cat.#: 64197, VWR
Chemicals, Radnor, PA, USA), acetonitrile (ACN, Cat.#: 75058, Fisher
Scientific, Waltham, MA, USA), chloroform (Cat.#: 67663, RCILabScan,
Bangkok, Thailand), 1,5-diaminonaphthalene (DAN, Cat.#: 56451, Sigma
Aldrich, St.Louis, MO, USA), 2′,5′-dihydroxyacetophenone
(DHA, Cat.#: D107603, Sigma Aldrich), ethanol (Cat.#: V002075; Sigma
Aldrich), formic acid (FA, Cat.#: 56302, Honeywell), and trifluoroacetic
acid (TFA, Cat.#: 40967; Honeywell, Charlotte, NC, USA). Luminescent
conjugated oligothiophene (LCO) tetramer formyl thiophene acetic acid
(q-FTAA) and heptamer formyl thiophene acetic acid (h-FTAA) were obtained
from Prof. Peter Nilsson, Department of Chemistry, Linköping
University. Water was obtained from a Synergy UV water purification
system (Milli-Q, Merck Millipore, Darmstadt, Germany).
5
Lipid Extraction and Characterization
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Materials: 1,5-diaminonaphthalene (DAN) and 2',5'dihydroxyacetophenone (DHA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). HPLC-grade acetonitrile, methanol, ethanol, chloroform, and tetrahydrofuran (THF) were purchased from Fisher Scientific (Pittsburgh, PA, USA). Lipid standards were purchased from Avanti Polar Lipids (Alabaster, AL, US).
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