Webinar: Laser Microdissection – Dissection Perfection

September 24, 2014

The two talks of this webinar are focussed on the practical advantages of this technique for a precise and contamination-free isolation and selection of single cells from brain sections for RNA downstream analysis and afterwards on cloning live cells from cultures for further cultivation and analysis.

Fig.: Lasermicrodissection of motor neurons from fresh frozen Nissl-stained adult mouse spinal cord tissue sections (for RNA analysis), using a Leica LMD7000 system.

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Laser Microdissection – Dissection Perfection

Laser microdissection is a microscope-controlled manipulation technique for the precise separation of samples using a guided focused laser beam. Today, it is an established method for a large number of applications, mainly in molecular biology, particularly nucleic acid research, cancer research, neurosciences, developmental biology, forensics, proteomics, plant research, for cutting live cell cultures and for single cell isolation. In this webinar the practical advantage of this technique for a precise and contamination-free isolation and the selection of single cells from brain sections and the use of laser microdissection to isolate distinct clones from live cell cultures for further cultivation (cloning) and additional analysis will be presented.

In particular the setting up of primary cultures from pancreatic adenocarcinoma is a delicate and complex procedure with very low chances of succeeding. One of the major issues is represented by the difficulty to separate epithelial cells from fibroblasts, which are very abundant due to the marked desmoplastic reaction associated to pancreatic adenocarcinoma. By laser microdissection It was be possible to isolate and to expand the epithelial cell population of primary cultures derived from pancreatic adenocarcinoma and use this effective models to evaluate gene expression of critical determinants of anticancer drug efficacy and predict different likelihood to respond to treatment.

In the field of neurodegenerative diseases there is now strong evidence that non-cell autonomous mechanisms can contribute to neuronal demise. Such pathological interactions between affected neurons and their surrounding glia and immune cells, called neuroinflammatory processes, can strongly influence disease progression, both in a beneficial or deleterious way. In the view of this theme, my research focuses on using mouse models to understand neurodegeneration in ALS (Amyotrophic Lateral Sclerosis) and in Parkinson’s disease. The two main questions Dr Christian Lobsiger asks are what are the stress and damage responses induced in the affected neuronal populations (motor neurons and dopaminergic neurons, respectively) and what are the corresponding responses induced in the surrounding glial/immune cell populations. We are using laser-microdissection (LMD) assisted RNA profiling to isolate micro-regions as well as neuronal or glial enriched pools to identify candidate pathways that could be implicated in these interactions. Targeting these pathways has the potential to modulate the ongoing neurodegenerative processes and slow disease.

Speakers:


Dr. Niccola Funel

Principal Investigator
Department of Surgical Pathology
University of Pisa


Dr. Christian Lobsiger

Team Member
Brain and Spinal Cord Institute
Salpetriere Hospital

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