Preparation of Hard Palm Seeds for Matrix-Assisted Laser Desorption/Ionization-Imaging Mass Spectrometry Analysis.


Journal

Journal of visualized experiments : JoVE
ISSN: 1940-087X
Titre abrégé: J Vis Exp
Pays: United States
ID NLM: 101313252

Informations de publication

Date de publication:
30 06 2023
Historique:
medline: 19 7 2023
pubmed: 17 7 2023
entrez: 17 7 2023
Statut: epublish

Résumé

Matrix-assisted laser desorption/ionization-imaging mass spectrometry (MALDI-IMS) is applied to identify compounds in their native environments. Currently, MALDI-IMS is frequently used in clinical analysis. Still, an excellent perspective exists for better applying this technique to understand chemical compounds' physiological information in plant tissues. However, preparation may be challenging for specific samples from botanical materials, as MALDI-IMS requires thin slices (12-20 µm) for appropriate data acquisition and successful analysis. In this sense, previously, we developed a sample preparation protocol to obtain thin sections of Euterpe oleracea (açaí palm) hard seeds, enabling their molecular mapping by MALDI-IMS. Here, we show that the developed protocol is suitable for preparing other seeds from the same genus. Briefly, the protocol was based on submerging the seeds in deionized water for 24 h, embedding samples with gelatin, and sectioning them in an acclimatized cryostat. Then, for matrix deposition, an xy motion platform was coupled to an electrospray ionization (ESI) needle spray using a 1:1 (v/v) 2,5-dihydroxybenzoic acid (DHB) and methanol solution with 0.1% trifluoroacetic acid at 30 mg/mL. E. precatoria and E. edulis seed data were processed using software to map their metabolite patterns. Hexose oligomers were mapped within sample slices to prove the adequacy of the protocol for those samples, as it is known that those seeds contain large amounts of mannan, a polymer of the hexose mannose. As a result, peaks of hexose oligomers, represented by [M + K]

Identifiants

pubmed: 37458441
doi: 10.3791/65650
doi:

Types de publication

Journal Article Video-Audio Media Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Auteurs

Gabriel R Martins (GR)

Laboratório de Biocatálise (LABIC), Instituto Nacional de Tecnologia; Departamento de Bioquímica, Instituto de Química, Universidade Federal do Rio de Janeiro.

Felipe L Brum (FL)

Laboratório de Biocatálise (LABIC), Instituto Nacional de Tecnologia; Departamento de Bioquímica, Instituto de Química, Universidade Federal do Rio de Janeiro; Centro de Espectrometria de Massas de Biomoléculas, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro.

Davi Marconi Miranda Carvalho da Silva (DMMC)

Laboratório de Biocatálise (LABIC), Instituto Nacional de Tecnologia; Departamento de Bioquímica, Instituto de Química, Universidade Federal do Rio de Janeiro.

Livia C Barbosa (LC)

Centro de Espectrometria de Massas de Biomoléculas, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro.

Ronaldo Mohana-Borges (R)

Centro de Espectrometria de Massas de Biomoléculas, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro.

Ayla Sant'Ana da Silva (AS)

Laboratório de Biocatálise (LABIC), Instituto Nacional de Tecnologia; Departamento de Bioquímica, Instituto de Química, Universidade Federal do Rio de Janeiro; ayla.santana@int.gov.br.

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Classifications MeSH