Laser Capture Microdissection: Methods for Microgenomic Analysis

Tue, Aug 4, 2009 - Fri, Aug 7, 2009
9:00 am - 5:00 pm

Trac 29 Laser Microdissection: Methods for Microgenomic Analysis

Tuesday, August 4 - Friday, August 7, 2009 • 9:00am – 5:00 pm
(28 hours of lecture and “hands-on” lab training)
Fee: $795 (Class limit of 24 participants)

This workshop, which is team taught by active researchers, will include lecture and hands-on laboratory exercises. Each participant will receive a comprehensive binder containing all material presented in the workshop along with laboratory protocols and reference material. There is a class limit of 24 participants with registration being on a first come first serve basis. 

Registration information can be obtained from Bea Sonnenberg at FAES (301-496-2316). Course schedule and content information can be obtained from Mark Nardone at 301-496-8290, nardonem@mail.nih.gov or from the Bio-Trac website at www.biotrac.com. If you desire further information on upcoming Bio-Trac workshops, please request by email and we can send you a pdf.

Laser Microdissection systems allows for the procurement of specific populations of cells from tissue and cytology samples containing heterogeneous populations of cells. The specificity of analyses are therefore much more representative of the disease process being studied. This approach to microdissection ensures that biological molecules, such as RNA and DNA, remain undamaged during the microdissection process. Downstream molecular analysis of these molecules produces accurate and assured results that have led to over 800 peer-reviewed publications by independent researchers.

In this four-day training program, participants learn to prepare tissue specimens for microdissection, then select and acquire homogenous cell populations using the mmi-CellCut, Leica LMD, Arcturus XT, Veritas and PALM microdissection systems. Instruction emphasizes operation of these LM systems, appropriate tissue handling and sample preparation for subsequent DNA, RNA or protein analysis, and methods for proper molecular extraction. Lecture and detailed instructions to prepare samples for several downstream molecular analyses are presented.

Topics:

 

Overview of Laser Microdissection Technology (History, Theory, Applications); Tissue Sample and Slide preparation; Project set up and QC of microdissected samples, Genomic and Epigenetic Analysis using PCR and microarray-based platforms, mRNA analysis including quantitative RT-PCR gene expression analysis from microdissected tissue samples, microRNA analysis from microdissected samples, and proteomic analysis from frozen and formalin-fixed, paraffin-embedded microdissected samples using Mass Spec. The lectures also include examples of scientific and clinical applications of LM-based studies. The hands on sessions include practice on Arcturus XT.

Course Agenda

Day 1: Tuesday August 4, 2009

Morning Lectures:
1.       Introduction to Laser Microdissection, the Course and the Technology (Jeffrey Hanson, NIH/NCI ) 9:00-10:00
2.       Pathology Review and Sample Preparation for Microdissection (Jaime Rodriguez-Canales, NIH/NCI)  10:05-11:05
3.       Project Design for Laser Microdissection (Jeffrey Hanson, NIH/NCI) 11:10-12:10

(Lunch 12:10-1:10)

Afternoon Lab Sessions:
4.       Lab Rotation I: Introduction to the Instruments (1:10-2:05)
5.       Lab Rotation II: Introduction to the Instruments (2:10-3:05)
6.       Lab Rotation III: Introduction to the Instruments (3:10-4:05)
7.       Lab Rotation IV: Introduction to the Instruments (4:10-5:05)
8.       Slide staining for Groups A and B, 5:10-6:00

Day 2: Wednesday August 5, 2009

Early Morning Lab Session:  Slide staining for Groups C and D, 8:00-8:50

Morning Lectures:
1.       DNA Studies from  Microdissected Samples (Steven Bova,  Johns Hopkins University) 9:00 to 10:00
2.       DNA Methylation of Microdissected Samples (Franziska Eberle,  NIH/NCI) 10:05 to 11:05
3.        Analysis of microRNAs from Microdissected Samples (Michael Tangrea, NIH/NCI) 11:10 to 12:10
4.       Panel Discussion 12:10-12:30

(Lunch 12:30-1:30)

Afternoon Lab Sessions:
5.       Lab Sessions,  1:30-5:30
a.       Lab Rotation 1: Hands on Laser Microdissection (1:30-3:10)
b.       Lab Rotation 2: Hands on Laser Microdissection (3:20-5:00)
c.       DNA extraction (5:00-5:30)

Day 3: Thursday August 6, 2009

Morning Lectures:
1.       RT-qPCR Analysis of Microdissected Samples,  (Heidi Erickson, MD Anderson Cancer Center), 9:00-10:00
2.       RNA from Microdissected Formalin Fixed Paraffin Embedded  Samples (Keith Steele, Maureen Kennedy, and Darci Smith, U.S. Army Medical Research Institute of Infectious Diseases (USAMRIID)) 10:05-11:05
3.       Gene Expression Analysis of Breast Tumor Microenvironment  (Stefan Ambs, NIH/NCI) 11:10-12:10
4.       Panel Discussion 12:10-12:30

(Lunch 12:30-1:30)

Afternoon Lab Sessions:
5.       Lab sessions 1:30-5:30
a.       Lab Rotation 3: Hands on Laser Microdissection (1:30-3:00)
b.       Lab Rotation 4: Hands on Laser Microdissection (3:00-4:30)
c.       DNA cleanup and  qPCR setup (4:30-5:30)

Day 4: Friday August 7, 2009

Special Session:  Analysis of the Lab Sessions Results: 8:20-8:50

Morning Lectures:
1.       Proteomic Analysis from Microdissected Samples using Mass Spectrometry (Donald Johann, NIH/NCI) 9:00-10:00
2.       Proteomic Analysis from FFPE Samples using Laser Microdissection (Vyomesh Patel, NIH/NIDCR) 10:05-11:05
3.       Laser Microdissection Applications  to Proteomics for Clinical Studies (Virginia Espina, George Mason University) 11:10-12:10
4.       Panel Discussion 12:10-12:30

(Lunch 12:30-1:30)

Afternoon Lab Sessions:
5.       Lab sessions (advanced Laser Microdissection Techniques) Sign up rotations 1:30-4:30 (three, one hour lab sessions)
6.       Optional  (NCI Laser Microdissection Core Demonstrations of PixCell II and Veritas Instruments) (for non local attendees) 5:45-7:00 (Building 10 / Room B1B37)

Optional Day: Monday August 10, 2009

Optional for in town attendees (NCI Laser Microdissection Core Demonstrations on Veritas and PixCell II instruments) 10:00-12:00

About Bio-Trac


Bio-Trac is an extensive series of post-graduate level "Hands On" biotechnology training courses given by the Foundation for Advanced Education in the Sciences (FAES) at the National Institutes of Health (NIH) in Bethesda, MD.
  • Team Taught by Active Researchers
  • Lecture and Hands-on Laboratory Training
  • Serving the Scientific Community for over 23 years
  • 11,500+ Scientists Trained
Sponsored by the Foundation for Advanced Education in the Sciences.