Infrared Laser Ablation Microsampling Coupled with MALDI Imaging

International Mass Spectrometry Conference, Florence, Italy, August 28, 2018

Louisiana State University: Kermit K. Murray, Fabrizio Donnarumma, Kelin Wang, Carson W. Szot, Chao Dong,
Baylor University: Touradj Solouki, Michael E. Pettit

Introduction
Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) is a powerful method for determining the location of biomolecules in tissue; however, protein identification and quantification remains challenging. The goal of this project is to develop an imaging workflow that combines MALDI imaging with laser ablation microsampling for liquid chromatography tandem mass spectrometry.

Methods
In the combined workflow, MALDI imaging is used to identify regions of interest (ROI) from intact proteins. The ROI are sampled using infrared laser ablation and the captured material is analyzed by LC-MS/MS using data independent acquisition to identify and quantify the proteins. The data are cross-correlated to identify the localized proteins in the MALDI images.

Results
Development of the combined approach is aimed at creating an automated system for ablation and capture and using it in a coupled workflow of MALDI imaging and LC MS/MS analysis. The infrared laser ablation and capture system uses a mid-infrared optical parametric oscillator laser with a custom reflective objective that has a large working distance and good numerical aperture. We have developed custom positioning software that allows MALDI MSI heat maps to be overlaid on camera images to co-register ROI ablation with the IR laser. Tissue sections are mounted on conductive microscope slides and either consecutive sections or MALDI analyzed sections can be used. Laser ablated proteins are digested with magnetic capture beads and the peptides released for analysis with a Waters nanoAcquity UPLC system coupled to a Synapt G2-HDMS.

Conclusions
MALDI MSI coupled with region-specific laser ablation sampling for LC MS/MS is a fast and versatile approach for spatially resolved tissue proteomics. We have demonstrated that proteins can be identified from spatially localized regions and are developing new methods for correlating the intact proteins observed in MALDI with the proteins identified by tandem mass spectrometry.

Novel Aspect: Coupled MALDI imaging with high precision infrared laser ablation capture for LC MS/MS for protein identification and quantification.

ASMS 2017

65th American Society for Mass Spectrometry Conference on Mass Spectrometry and Allied Topics, Indianapolis Indiana, June 4 – 8, 2017

Downtown Indianapolis
Downtown Indianapolis
Sunday Tutorial

MALDI – Past and Future, Kermit K. Murray

Monday Poster 644

Combined Proteomic and RNA Expression Analysis of Laser Ablation Extracted Tissue, Chao Dong, Kelin Wang, Fabrizio Donnarumma, Scott W. Herke, Michael E. Pettit, Touradj Solouki,3 and Kermit K. Murray, presented at the 65th ASMS Conference on Mass Spectrometry and Allied Topics, Indianapolis, Indiana, June 5, 2017.

Combined Proteomic and RNA Expression Analysis of Laser Ablation Extracted Tissue
Combined Proteomic and RNA Expression Analysis of Laser Ablation Extracted Tissue
Chao Dong ASMS 2017
Chao Dong ASMS 2017
Monday Poster 698

Infrared Laser Ablation Sample Transfer with Conserved Biological Function, Kelin Wang, Matthew D. Baldone, Fabrizio Donnarumma, and Kermit K. Murray, presented at the 65th ASMS Conference on Mass Spectrometry and Allied Topics, Indianapolis, Indiana, June 5, 2017.

Infrared Laser Ablation Sample Transfer with Conserved Biological Function
Infrared Laser Ablation Sample Transfer with Conserved Biological Function

Kelin MP698 2017

Tuesday Poster 429

Pulsed Valve Matrix-assisted Ionization, Bijay Banstola and Kermit K. Murray, presented at the 65th ASMS Conference on Mass Spectrometry and Allied Topics, Indianapolis, Indiana, June 6, 2017.

Pulsed Valve Matrix-assisted Ionization
Pulsed Valve Matrix-assisted Ionization
Bijay Banstola ASMS 2017
Bijay Banstola ASMS 2017
Tuesday Poster 500

Combined Solid and Liquid Matrix High Vacuum Sublimation and Evaporation Coating for MALDI Imaging, Fan Cao, Fabrizio Donnarumma, Md Amir Hossen, and Kermit K. Murray, presented at the 65th ASMS Conference on Mass Spectrometry and Allied Topics, Indianapolis, Indiana, June 6, 2017.

Combined Solid and Liquid Matrix High Vacuum Sublimation and Evaporation Coating for MALDI Imaging
Combined Solid and Liquid Matrix High Vacuum Sublimation and Evaporation Coating for MALDI Imaging
Amir Hossen ASMS 2017
Amir Hossen ASMS 2017
Wednesday Poster 362

Online Atomic Force Microscope Tip-enhanced Laser Ablation Mass Spectrometry, Md Amir Hossen, Bijay Banstola, Fan Cao, and Kermit K. Murray, presented at the 65th ASMS Conference on Mass Spectrometry and Allied Topics, Indianapolis, Indiana, June 6, 2017.

Online Atomic Force Microscope Tip-enhanced Laser Ablation Mass Spectrometry
Online Atomic Force Microscope Tip-enhanced Laser Ablation Mass Spectrometry
Remi Lewal, Chao Dong, Kelin Wand, Fan Cao, and Bijay Banstola at ASMS 2017
Remi Lewal, Chao Dong, Kelin Wand, Fan Cao, and Bijay Banstola at ASMS 2017
Thursday Poster 260

Infrared Laser Ablation Microsampling and Data Independent Acquisition Mass Spectrometry for Tissue Proteomics, Kelin Wang, Fabrizio Donnarumma, Carson W .Szot, Michael E. Pettit, Touradj Solouki, and Kermit K. Murray

Infrared Laser Ablation Microsampling and Data Independent Acquisition Mass Spectrometry for Tissue Proteomics
Infrared Laser Ablation Microsampling and Data Independent Acquisition Mass Spectrometry for Tissue Proteomics
Kelin Wang ASMS 2017
Kelin Wang ASMS 2017
Thursday Poster 695

Laser Ablation Capture of Formalin Fixed Paraffin Embedded Tissue for LC-MS/MS Proteomics Analysis, Fabrizio Donnarumma, Michael E. Pettit, Touradj Solouki, and Kermit K. Murray

Laser Ablation Capture of Formalin Fixed Paraffin Embedded Tissue for LC-MS/MS Proteomics Analysis
Laser Ablation Capture of Formalin Fixed Paraffin Embedded Tissue for LC-MS/MS Proteomics Analysis
Fabrizio Donnarumma ASMS 2017
Fabrizio Donnarumma ASMS 2017
Kelin Wang and Fan Cao at ASMS 2017
Kelin Wang and Fan Cao at ASMS 2017
Murray Group Dinner ASMS 2017: Amir Hossen, Chao Dong, Remi Lewal, Kelin Wang, Fan Cao, Fabrizio Donnarumma, Bijay Banstola, Sucharita Dutta, Shelley Jackson, Kermit Murray, Jeonghoon Lee, and Xing Fan
Murray Group Dinner ASMS 2017: Amir Hossen, Chao Dong, Remi Lewal, Kelin Wang, Fan Cao, Fabrizio Donnarumma, Bijay Banstola, Sucharita Dutta, Shelley Jackson, Kermit Murray, Jeonghoon Lee, and Xing Fan
ASMS Livery Indy Car
ASMS Livery Indy Car
Indiana State House at ASMS 2017
Indiana State House at ASMS 2017

ASMS 2007: Laser Desorption Electrospray Ionization

J. Dong, Y. H. Rezenom, and K. K. Murray, “Aerosol Desorption Electrospray Ionization,” Presented at the 55th ASMS Conference on Mass Spectrometry, June 4, 2007, Indianapolis, Indiana, Ambient Ionization I, MP 006.

2007 ASMS: Aerosol Desorption Electrospray Ionization

American Society for Mass Spectrometry Workshop “Infrared Lasers for MALDI” June 4, 2007


IR Laser + Electrospray Instrument Photos April 16, 2007