Infrared Laser Ablation Microsampling for Small Volume Proteomics.
J Am Soc Mass Spectrom
; 33(6): 1003-1010, 2022 Jun 01.
Article
in En
| MEDLINE
| ID: mdl-35536596
ABSTRACT
Infrared (IR) laser ablation was used to remove localized tissue regions from which proteins were extracted and processed with a low volume sample preparation workflow for bottom-up proteomics by liquid chromatography tandem mass spectrometry (LC-MS/MS). A polytetrafluoroethylene (PTFE) coated glass slide with 2 mm diameter microwells was used to capture ablated rat brain tissue for in situ protein digestion with submicroliter solution volumes. The resulting peptides were analyzed with LC-MS/MS for protein identification and label-free quantification. The method was used to identify an average of 600, 1350, and 1900 proteins from ablation areas of 0.01, 0.04, and 0.1 mm2, respectively, from a 50 µm thick rat brain tissue section. Differential proteomics of 0.01 mm2 regions captured from cerebral cortex and corpus callosum was accomplished to demonstrate the capabilities of the approach.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Proteomics
/
Laser Therapy
Limits:
Animals
Language:
En
Journal:
J Am Soc Mass Spectrom
Year:
2022
Document type:
Article
Affiliation country:
Estados Unidos