A Schwarzschild reflective objective with a numerical aperture of 0.3 and working distance of 10 cm was used for laser ablation sampling of tissue for off-line mass spectrometry. The objective focused the laser to a diameter of 5 μm and produced 10 μm ablation spots on thin ink films and tissue sections. Rat brain tissue sections 50 μm thick were ablated in transmission geometry, and the ablated material was captured in a microcentrifuge tube containing solvent. Proteins from ablated tissue sections were quantified with a Bradford assay, which indicated that approximately 300 ng of protein was captured from a 1 mm area of ablated tissue. Areas of tissue ranging from 0.01 to 1 mm were ablated and captured for bottom-up proteomics. Proteins were extracted from the captured tissue and digested for liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis for peptide and protein identification.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8895455 | PMC |
| http://dx.doi.org/10.1021/jasms.1c00306 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Immunohistochemical labeling of ongoing axonal degeneration 10 days following cervical contusion spinal cord injury in the rat.
Spinal Cord
January 2025
McKnight Brain Institute, University of Florida, Gainesville, FL, USA.
Study Design: Experimental Animal Study.
Objective: To continue validating an antibody which targets an epitope of neurofilament light chain (NF-L) only available during neurodegeneration and to utilize the antibody to describe the pattern of axonal degeneration 10 days post-unilateral C4 contusion in the rat.
Setting: University of Florida laboratory in Gainesville, USA.
Fracture properties of porcine versus human thoracic aortas from tricuspid/bicuspid aortic valve patients via symmetry-constraint Compact Tension testing.
Sci Rep
January 2025
Department of Engineering Mechanics, KTH Royal Institute of Technology, Stockholm, Sweden.
Aneurysm rupture is a life-threatening event, yet its underlying mechanisms remain largely unclear. This study investigated the fracture properties of the thoracic aneurysmatic aorta (TAA) using the symmetry-constraint Compact Tension (symconCT) test and compared results to native and enzymatic-treated porcine aortas' tests. With age, the aortic stiffness increased, and tissues ruptured at lower fracture energy [Formula: see text].
View Article and Find Full Text PDFSequential MALDI-HiPLEX-IHC and Untargeted Spatial Proteomics Mass Spectrometry Imaging to Detect Proteomic Alterations Associated with Tumour Infiltrating Lymphocytes.
J Proteome Res
January 2025
Department of Medicine and Surgery, Proteomics and Metabolomics Unit, University of Milano-Bicocca, Vedano al Lambro 20854, Italy.
MALDI-HiPLEX-IHC mass spectrometry imaging (MSI) represents a newly established workflow to map tens of antibodies linked to photocleavable mass tags (PC-MTs), which report the distribution of antigens in formalin-fixed paraffin-embedded (FFPE) tissue sections. While this highly multiplexed approach has previously been integrated with untargeted methods, the possibility of mapping target cell antigens and performing bottom-up spatial proteomics on the same tissue section has yet to be explored. This proof-of-concept study presents a novel workflow combining MALDI-HiPLEX-IHC with untargeted spatial proteomics to analyze a single FFPE tissue section, using clinical clear cell renal cell carcinoma (ccRCC) tissue as a model.
View Article and Find Full Text PDFDecellularization and Enzymatic Digestion Methods to Enhance ECM Protein Detection via MALDI-MS Imaging.
Anal Chem
January 2025
Molecular and Cellular Biology Graduate Program, University of Massachusetts, Amherst, Massachusetts 01003, United States.
Matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI-MSI) has been used to generate spatial maps of lipids, metabolites, peptides, proteins, and glycans in tissues; however, its use for mapping extracellular matrix (ECM) protein distributions is underexplored. ECM proteins play a major role in various pathological conditions, and changes in their spatial distributions affect the function and morphology of cells within tissues. ECM protein detection is challenging because they are large, insoluble, and undergo various post-translational modifications, such as glycosylation.
View Article and Find Full Text PDFEfficacy of PDD-guided tumor excision combined with photodynamic therapy in cutaneous squamous cell carcinoma.
Photodiagnosis Photodyn Ther
January 2025
Department of Plastic & Cosmetic Surgery, Daping Hospital, Army Medical University, Chongqing 400042, China. Electronic address:
Background: Cutaneous squamous cell carcinoma (cSCC) is an aggressive tumor with unclear margins. Photodynamic diagnosis (PDD) enables the differentiation of tumor tissue from normal tissue via visible fluorescence. Photodynamic therapy (PDT) is widely used in the treatment of non-melanoma skin tumors.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!