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Nanyang Technological University

Laser Microdissection-Based Protocol for the LC-MS/MS Analysis of the Proteomic Profile of Neuromelanin Granules

2.2K Views

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07:35 min

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December 16th, 2021

DOI :

10.3791/63289-v

December 16th, 2021

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Maximilian Wulf¹^,², Katalin Barkovits-Boeddinghaus¹^,², Paula Sommer¹^,², Karin Schork¹^,², Martin Eisenacher¹^,², Peter Riederer³^,⁴, Manfred Gerlach⁵, Steffen Kösters¹^,², Britta Eggers*¹^,², Katrin Marcus*¹^,²

¹Medizinisches Proteom-Center, Medical Faculty, Ruhr-University Bochum, ²Medical Proteome Analysis, Center for Proteindiagnostics (PRODI), Ruhr-University Bochum, ³Center of Mental Health; Clinic and Policlinic for Psychiatry, Psychosomatics and Psychotherapy, University Hospital Wuerzburg, ⁴Psychiatry Department of Clinical Research, University of Southern Denmark Odense University Hospital, ⁵Center of Mental Health, Department of Child and Adolescent Psychiatry, Psychosomatics and Psychotherapy, University Hospital of Wuerzburg, University of Wuerzburg

Summary

A robust protocol is presented here for isolating neuromelanin granules from human post-mortem substantia nigra pars compacta tissue via laser microdissection. This revised and optimized protocol massively minimizes the required time for sample collection, reduces the required sample amount, and enhances the identification and quantification of proteins by LC-MS/MS analysis.

Explore More Videos

Laser Microdissection

LC MS MS Analysis

Proteomic Profile

Neuromelanin Granules

Spatial Resolution

Tissue Analysis

Postmortem Brain Tissue

Sample Isolation

Disease Mechanisms

Field Of View Analysis

Sample Collection

Triptic Digestion

HPLC MS Analysis

Chapters in this video

0:05

Introduction

0:46

Laser Microdissection and Pressure Catapulting

2:46

Sample Preparation for HPLC-MS and Proteomic Raw Data Analysis

5:10

Statistical Analysis

5:50

Results: Protein Identification in Neuromelanin Granules (NMGs) and Substantia nigra (SN)

7:00

Conclusion

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