Applications Leica LMD7000

Application Notes include short descriptions of research experiments.

Please convince yourself on versatility of Leica LMD Laser Microdissection system.

• Gene Expression Analysis, EN (0,18 MB)
• Live Cell Cutting, EN (0,32 MB)
• Plant Root Meristem, EN (0,51 MB)
• Protein Analysis, EN (0,16 MB)
• Quantitative RT PCR, EN (0,34 MB)
• Spermatozoa Forensic, EN (0,21 MB)
• Tooth Tissue, EN (0,22 MB)
• Laser Microdissection Objectives, EN(0,94 MB)
• AVC Software LMD6000, EN (2,01 MB)
• DNA Damage and Repair, EN (0,88 MB)
• NEW - Plant Research, EN (0,27 MB)

LMD applications require special slides, which are specifically designed to free the dissectate from the section.

For this reason, tissue sections to be microdissected are placed on a UV-absorbing membrane.

Upon the laser action, the membrane is vaporized, which also vaporizes the tissue in the cut line. Since the membrane is not adhered to the slide, the dissectate (with the attached specimen) drops into the cap of the microcentrifuge tube. Leica Microsystems offers high-quality membrane slides that provide high collection rates for laser microdissection.

There are three different membrane slides to choose from:

• PEN (polyethylene naphthalate)
• PET (polyethylene terephthalate)
• POL (polyester)

Membranes are mounted on regular glass slides (PEN), on steel frames (PET/POL), or on the bottom of special Petri dishes for cell cultivation. Depending on tissue type, thickness, and inspection method, one slide type may work better for a certain application. Therefore it is advisable to test different combinations of membranes and slide supports for the best results. Membrane combinations are summarized in a Table, click link above "Table Slides and Petri dishes".

Glass PEN foil slide (upper image on top) and the frame slide (upper image bottom).
Description: The membrane is mounted on the upper side of the slide (the side with the frosted end for marking). Since the membrane is glued on the edges in thecentral part of the slide, there is a space of 1 µm between the glass and membrane. The slides must be handled with care to prevent damage to the membrane, this will impede the laser action.

Petri dish for cell cultivation for LMD (right image).
Description: The Petri dish is placed on the holder for firm mounting on the microscope stage. Since the PEN membrane is perforated, the cells are perfectly supplied with oxygen. For better attachment and growth of cells, the membrane can be coated with poly-Lysine or collagen prior to seeding.

The frame slide (upper image on top) and the frame support (upper image on bottom).
Description of the frame slide: The front side of the slide contains the engraved Leica logo. Please note that only the back side of the slide (the side opposite the Leica logo) is foil mounted, and only this side can be used for mounting tissue sections.
Description of the frame support: The Plexiglas frame support contains the lifted surface (clear area) that fits exactly into the cavity or the front side of the frame slide. Not shown, the membrane slide must be turned over to be mounted onto the frame support.

Important: Please note that sterilizing slides by autoclaving or UV-treatment does not guarantee complete destruction of RNases. For RNA preparation, especially from a single cell (corresponds in average to 10-15 pg of total RNA) or from a small number of cells, RNase-free certified slides are recommended (see Table 1). Or alternatively, regular glass slides can be pretreated with reagents such as RNase Zap® (Ambion).

Citation list for Leica LMD Laser Microdissection

Please click here to open the pdf file with the citation list for Leica LMD Laser Microdissection.

One Solution – Two Competent Partners

Leica is a leading supplier of high precision optical microscopes and related instruments, and provides efficient tools for all the steps required for laser microdissection of tissue samples.
QIAGEN has a worldwide reputation for its technologies for nucleic acid purification, and provides optimized reagents and protocols for purification and analysis of nucleic acids from dissected samples.

The agreement between QIAGEN and Leica combines the expertise of two leading companies to bring you complete solutions for analyzing nucleic acids from laser microdissected specimens.

Leica Qiagen Brochure

Visit Qiagen

Reliable instruments for infiltration, embedding and sectioning

Prepare tissue sections by using a variety of sectioning techniques. Choose the appropriate Leica instrument for your application: cut frozen tissue with a cryostat, fixed and paraffinembedded tissues using the Leica RM22 family of rotary microtomes, or native, unfrozen tissue under physiological conditions with a Leica VT1200/1200 S vibrating microtome.

Efficient staining of tissue sections

Stain your fixed sections and visualize cell structures at high resolutions. Staining workstations from Leica allow several staining protocols to run simultaneously, and can be programmed to satisfy customized requirements. Built-in filters reduce exposure to hazardous fumes, and the staining process can be monitored through a transparent lid.

Experience a New Level of Speed

Safely and accurately, capture single cell(s) and tissue regions of any shape or size. The Leica LMD7000 allows you to easily collect samples with a fast, precise, contamination-free method - simply by gravity.
The Leica LMD7000 is a fully automated laser microdissection system based on the Leica DM6000 B. This automation allows you to harvest one or more cell types from a single slide directly into one or more PCR tubes.

Excise cells or metaphase chromosomes from a wide range of samples, including paraffin-embedded sections, frozen sections, blood smears, and stained and immunolabeled specimens.
The Leica LMD7000, was developed combining automated upright microscope architecture, three-dimensional optical control of the dissecting laser beam and the dissected area, non-contact tissue sampling and motorized post-dissection handling.

Efficient purification of DNA, RNA, miRNA, and proteins

Purify analytes from laser-microdissected tissue sections using QIAGEN® Sample Technologies that are specially designed to process very small amounts of starting material.
Efficient purification of DNA and RNA from laser-microdissected fresh-frozen tissues is achieved using the QIAamp® DNA Micro Kit (cat. no. 56304) and RNeasy® Plus Micro Kit (cat. no. 74034), espectively.

For lasermicrodissected formalin-fixed paraffin-embedded (FFPE) tissues, the QIAamp DNA FFPE Tissue Kit (cat. no. 56404), RNeasy FFPE Kit (cat. no. 74404), and miRNeasy FFPE Kit (cat. no. 217404) maximize recovery of usable DNA, RNA, and miRNA, respectively. Full-length proteins can also be extracted from whole FFPE tissue sections using the Qproteom^TM FFPE Tissue Kit (cat. no. 37623).

Highly specific and sensitive amplification for detailed characterization of nucleic acids

Amplify the nucleic acids purified from laser-microdissected tissue sections using QIAGEN Assay Technologies, which are ideally suited for very small amounts of starting material. Highly specific hot-start PCR solutions such as HotStarTaq® Plus DNA Polymerase (cat. no. 203603) and the QIAGEN Multiplex PCR Kit (cat. no. 206143) facilitate fast and successful molecular analysis of DNA.

The QuantiTect® Reverse Transcription Kit (cat. no. 205311) is optimized for cDNA synthesis with integrated genomic DNA removal from very small amounts of RNA (10 pg to 1 μg). Quantitative, real-time PCR and RT-PCR with QuantiTect Multiplex PCR and RT-PCR Kits (cat. nos. 204543 and 204643) allow analysis of multiple targets (2–4) in 1 reaction vessel on any commercially available real-time PCR cycler. All QIAGEN PCR and RT-PCR products include optimized buffers that enhance PCR specificity and minimize optimization of reaction parameters, enabling fast and successful results at the first attempt.

The objectives described on the attached files are specifically intended for laser microdissection. They are designed for high UV-light transmission upon laser cutting, and to provide the best image quality.

The Leica SmartCut Series comprises the following objectives for LMD: UVI 6.3x and HCX PL FLUOTAR family (20x, 40x, 63x, and 150x).

Improved optics provide high performance for laser microdissection of single cells as well as larger tissue areas.

• Application Notes LMD Objectives, EN(0,94 MB)
• Table SmartCut LMD Objectives (0,03 MB)