A non-instrument-based method for the analysis of formalin-fixed paraffin-embedded human spinal cord via matrix-assisted laser desorption/ionisation imaging mass spectrometry

Rapid Communications in Mass Spectrometry : RCM
Matthew B O'RourkeMatthew P Padula

Abstract

This paper, in conjunction with a work published earlier this year by O'Rourke et al., aims to provide a comprehensive set of protocols for the analysis of formalin-fixed paraffin-embedded (FFPE) tissue via matrix-assisted laser desorption/ionisation imaging mass spectrometry (MALDI IMS) in a low-cost and highly repeatable and robust way, thereby allowing other research teams to begin their own IMS-centered avenues of research. Samples of FFPE tissue were sectioned at 5 µm, water float mounted to specially prepared ITO glass slides and then dilipidated in a graded alcohol series. Tissue sections were then antigen retrieved under pressure in 20 mmol Tris-HCl (pH 8.8), coated with trypsin and digested O/N at 37°C. Samples were then sublimated with matrix to a final coverage of 0.2 mg/cm(2), recrystallised at 37°C with 50:50 acetonitrile (ACN)/0.1% trifluoroacetic acid (TFA) for 1 h and analysed with a MALDI TOF/TOF mass spectrometer. Serial sections were imaged, revealing little to no variation with regards to image quality and corresponding spectra. We have also attempted to describe the processes that govern the various aspects of this protocol with respect to each step necessary to ensure reproducibility. We are confident that...Continue Reading

References

Sep 1, 1996·Biotechnic & Histochemistry : Official Publication of the Biological Stain Commission·C R TaylorR J Cote
Feb 18, 2004·Laboratory Investigation; a Journal of Technical Methods and Pathology·Vladimir K RaitJeffrey T Mason
Sep 13, 2005·Laboratory Investigation; a Journal of Technical Methods and Pathology·David K CrockettKojo S J Elenitoba-Johnson
Oct 10, 2009·Current Protocols in Molecular Biology·Rebekah L GundryJennifer E Van Eyk
Apr 20, 2010·Brain Research·Catherine Anne GorriePhil M E Waite
Jan 23, 2013·Proteomics. Clinical Applications·Shan-Rong ShiJeffrey T Mason
Apr 11, 2013·Advanced Drug Delivery Reviews·Eric M Weaver, Amanda B Hummon
May 9, 2013·Histochemistry and Cell Biology·Andreas Römpp, Bernhard Spengler
Jun 14, 2013·International Journal of Molecular Sciences·Surendra NimeshPrem Kumarathasan
Apr 2, 2014·Journal of Proteomics·Megan M GesselRichard M Caprioli

❮ Previous
Next ❯

Citations

Jul 16, 2016·Mass Spectrometry Reviews·Matthew B O'RourkeMatthew P Padula
Jan 10, 2018·Journal of the American Society for Mass Spectrometry·Matthew B O'RourkeMatthew P Padula
Feb 9, 2019·Current Protocols in Molecular Biology·Matthew B O'RourkeMatthew P Padula
Nov 8, 2019·Journal of Mass Spectrometry : JMS·Matthew B O'Rourke, Matthew P Padula
Jan 29, 2020·Analytical and Bioanalytical Chemistry·Juliane HermannJoachim Jankowski
Oct 13, 2016·Proteomics. Clinical Applications·Matthew B O'Rourke, Matthew P Padula
Nov 27, 2020·Proteomes·Matthew B O'Rourke, Matthew P Padula
Feb 4, 2021·Analytical and Bioanalytical Chemistry·Angus C GreyNicholas J Demarais

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.