Molecular Machines & Industries

Laser Microdissection System

mmi CellCut Plus

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NEW-GENERATION TECHNOLOGYMMI is leading the way to provide scientists with innovative and sophisticated, solid state laser capture microdissection in-struments. We are aiming to make this core technology in many scientific fields to be more efficient and cost effective. The patented adhesive cap technology is reliable, efficient and easy to use.

The modular concept of the MMI instruments offers a highly flexible instrument platform wherein optical tweezers, mechani-cal micromanipulation, fluorescence and other modules can be added to meet and surpass your requirements. The biggest benefit for our customers is an instrument that is easy to use and capable of performing the most complex research.

Today, more than ever, it is MMI‘s goal to provide you with the highest degree of precision possible. Our technology keeps your samples contamination free and your experimental results reproducible.

Our application support and service team is well-trained and experienced. They will help you to achieve your goals in opti-mizing your experiments to get accurate results.

The MMI team is committed to our customers and we stand behind our products. That is why we invite you to discover and innovate with us.

LEAD THE WAY to a new level of microdissection.

Sincerely yours,The MMI Team

All rights reserved at MMI AG, 2009

LEAD THE WAY to a new generation LCM technology

FIRST IN CUTTING-EDGE TECHNOLOGYThe mmi CellCut Plus system is the ultimate laser microdissection tool forresearchers who effortlessy wanted to isolate groups of cells, single cells and cell components for analysis across a wide range of applications.

mmi CellCut Plus is ideal for scientists interested in the following areas of research: molecular/cell biology, genomics and proteomics, forensics, can-cer and stem cell research. Clinically related applications such as molecular pathology, microbiology, and virology will also find the mmi CellCut Plus of primary importance and value.

Principles of laser microdissection

mmi CellCut Plus combines several proven, leading-edge technologies in providing an extremely fast, precise and clean isolation of cells from a wide range of microscopic sam-ples. This is achieved by dissecting the areas of interest with an ultra precise UV-laser while maintaining it‘s morphology and ensuring the quality of the source material for subsequent downstream analysis without adding of physical or chemical forces.

High-speed ultra fine laser cutting

The mmi CellCut Plus system is fully controlled through the easy-to-use mmi CellTools soft-ware. This user-friendly program provides a live view of the microscopic sample on the monitor and allows the user to identify, mark and isolate the areas of interest easily.

For cell isolation, the maintenance-free, solid-state UV-laser is focused through the microscope’s objective onto a microscopic sample to enable a cut width of as small as 0.3μm with the 100x objective. This, combined with a very short (pico-second) pulse duration and a high repetition rate, provides an ultra precise and fast target excision. The mmi CellTools software controls the focus and energy of the laser in microdissecting the area of interest while the laser remains aligned at the center. The highly precise motorized stage is used to accurately move the sample on the inverted research microscope.

Due to the very low (< 1 μjoule) pulse energy of the laser used in the mmi CellCut Plus sys-tem, subsequent molecules like DNA, RNA and protein is unaffected by the laser dissection process. This means that cells and cell components from frozen- and/or paraffin-embedded tissues, archived material, smears, cytospins, as well as living cells from cell cultures can be marked on the display of the microscope’s image and microdissected without negative impact on the quality of the material for downstream applications.

Contamination-Free Isolation technology

The unique mmi CapLift technology provides an automated transfer of uncontaminated targets from the entire microscope slide into micro centrifuge tubes. The captured samples adhere to the mmi IsolationCap of the extraction tube, which snaps close to keep the sample free from contamination and ready for downstream molecular biological analysis.

Key customer benefits:

• Ultra high precision laser cutting for best laser microdissection results• Ultra-short pulse, low average power laser for best DNA/RNA/Protein

quality recovery• Maintenance free and certified long lifespan laser• Retained orientation of microdissection samples to enable scientists to

view the cells exactly as they appear on the tissue section• Regions of interest can be inspected and processed again after micro-

dissection

Computer-controlled movements

UV-Cut software-controlled laser focus and energy

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A Principle of microdissectionfast, precise and contamination-free

B mmi CapLift technology

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Application examples

Multiple cells in pathological samples

Differences in genomic (DNA/RNA) and proteomic expression of any tissue type can be analyzed easily. For example, transverse sections of intestinal glands from colon tissues can be identified and quickly isolated for study of specific genes and the corresponding hormone response.

Single cells in hematology or cytology

Single cells, such as a typical plasmoblast from a blood smear or other relevant cytological cells, can be identified, cut, and isolated with the mmi CellCut Plus system. With the remai-ning sample securely attached to the mmi IsolationCap, it is easy to match the isolated cell and the remaining sample in documenting the precision and the efficiency of the extraction.

Fluorescence-labeled cell components

The precision of the mmi CellCut Plus system also enables you to simply locate and cleanly microdissect structures smaller than single cells. For example, chromosomes with detectable fluorescence in situ hybridization (FISH) signals can be isolated for subsequent investigation.

Forensic applications

The mmi CellCut Plus system in forensic medicine is used, among other applications to isolate a single sperm from a vaginal smear for genetic analysis, greatly increasing the possibility of a positive match and therefore possibly leading to a criminal conviction.

Other samples

Living specimens like C. elegans can also be examined with the mmi CellCut Plus sys-tem. Here, the living organism can be placed or fixed, between the membrane and a glass slide, in order to isolate and extract areas of interest as required by the individual experi-ment.

Live cells

Most stem cell lines are presently grown at high densities on mouse fibroblast “feeder lay-ers”. Therefore, isolating specific pluripotent cells from the surrounding fibroblasts and diffe-rentiating cells needs to be fast and precise. With mmi CellCut Plus, there is a perfect balan-ce between speed and precision, allowing the target stem cells to be isolated and re-cultured without any side effects, such as karyotype changes.

Basic laser micromanipulation

The mmi CellCut Plus system can also be used as a laser micromanipulation system. Single, short laser pulses produce small self-sealing holes in the plasma membrane of a living sample, which improves protoplast fusion or increases the transfection rates of exogenous substances.

Proteomic Applications

In proteomic research, several thousand cells are needed to run Western-blot or two di-mensional gels. Using the mmi CellCut Plus enables scientists to quickly microdissect cells of interest without any potential damage, as compared to alternative methods like polymer thermo-excitation or cell projection.

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Fluorescence stained chromosomesC

Blood smear with a typical plasmoplastB

Single sperm isolated from a gynaecological smear

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Live C. elegans fixed onto a membrane and isolated.

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Tissue sections of colon

LEAD THE WAY to a wide range of applications

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LEAD THE WAY to a quick and clean isolation

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Microdissection of fluorescence stained chromosomes*

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Blood smear with atypical plasmoblasts*

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Single sperm isolated from a gynaeco-logical smear

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Live C. elegans, fixed onto a membrane and isolated.

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Intestinal gland isolated from transverse tissue sections of colon*

Cryo or paraffin-preserved tissue

Chromosomes

Others, e.g. sperms, C. elegans

Selected targetIncubation and centrifugationI

H Easy cell selection

Assembly of the reversed metal frame and a glass slide (1.0 or 0.17 mm)

Unique sandwich with contamination- free and protected sample

QUICK AND CLEAN TARGET ISOLATIONUnique system for sample preparation and sample protection

For the sample preparation of any source such as frozen or paraffin embed-ded, smears or cytospins and chromosomes spreads, the mmi CellCut Plus sys-tem uses the mmi MembraneSlides. This special frame slide is covered with a thin membrane that is completely inert and has negligible auto fluorescence. The diffe-rent types of samples are prepared on this membrane and are covered with a normal glass slide for protection against contamination.

Selection and cutting

Using the mmi CellCut Plus software, microdissecting the regions of interests are se-lected on the display either by using the mouse freehand or predefining geometrical shapes, such as circles and squares. Any number of areas across the entire slide can be identified as targets and the sizes of the geometrical shapes can be changed as well as copied and pasted for consistency.

Fast, precise and clean excision

The thin (0.3 µm at 100x) cutting path enables a precise and comfortable extraction of the selected areas at an outstanding speed without affecting its morphology or otherwise, negatively affecting the areas of interests. As a result, there is no loss in quality of the material used in subsequent steps. Even the viability of living cells is not affected and therefore, the cells, once selected, can be re-cultured. Depending on the sample type, several thousands of cells can be laser dissected under a minute.

Ready for “downstream” analysis

The mmi IsolationCaps used in the mmi CapLift technology allows the collection of tar-get areas across the entire microscope slide. After microdissection, the mmi IsolationCaps are snapped into the micro centrifuge tubes to undergo extraction of the bio molecules. After using the recommended extraction reagents and at the desired incubation time, the extracted targets are now ready for further genomic and proteomic processing.

Key customer benefits

• Unique mmi sample preparation and protection system allowing con-tamination-free working conditions (RNAse/DNase free and safe from environmental contamination)

• Simple and comfortable working processes for more efficient results• The only system which allows reworking already isolated areas without

the need for a larger area ablation• Best results are even achieved from limited source of material; optimi-

zing technology to ensure DNA and RNA quality• Keeps you in control of your research work without compromising

speed and efficiency.

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F Sample preparationon frame slide with PET or PEN membrane

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Single cells, smears or cytospins

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MMI Live Cell Microdissection –The new mmi Live CellChamberIntroduction: The isolation and enrichment of individual live cells for culture anddifferentiation experiments or for their proteomic and genomic analysis is of increasing interest in stem cell research, cancer research and tissue engineering. While mechanical separation techniques are cumbersome, time consuming and bear the risks of contami-nation or mechanical stress, faster Laser Microdissection based methods are current-ly under development. The mmi CellCut Plus in combination with the newly developed mmi Live CellChamber enables contamination-free isolation of cells in living culture.

Components of the new mmi Live CellChamber

• A membrane ring, for the initial cultivation of cells• A cell culture dish, to house the membrane ring with seeded cells• The microdissection chamber, UV – permeable and filled with adhesive for Laser Microdissection of single cells or group of cells.

Note: All components are sterilized and ready to use.

The Workflow

Adherent cells are grown in a special metal ring on a membrane inside a petri dish. Once a desired cell density is reached, the membrane ring is transferred to the adhesive area of the microdissection chamber. The cell or cells of interest are selected and cut using Laser Microdissection. In addition, unwanted cells can be destroyed or ablated by using individu-al laser shots. The cell chamber is then removed and only the cells of interest remain in the microdissection chamber.

Repeated selection of wanted cells: This process can be repeated using the same metal ring in a new microdissection chamber to select and separate more than one cell type. Once isolation is complete, sufficient medium is added to the microdissection chamber and the cells can be re-cultivated.

Seed cells and cultivate to the desired cell density. Visualize your cells of interest (e.g. phase contrast, immuno labeling, etc.)

Transfer the membrane ring with cells into the mmi microdissection chamber

Selection of cells via LMD

Isolate cells by removing the membrane ring with unwanted cells. Reculture the cells of interest and if desired, the depleted fraction.

Repeat if desired

Your Advantage

The isolation of single cells is a unique and highly useful tool in wide range of research and clinical applications, including individual cell analysis, clonal expansion, transfection expe-riments and stem cell research. Enzyme treatments (i.e. trypsin), other potentially harmful selective reagents, as well as time consuming repetitive enrichments can be avoided. The benefit is an increase in reliability, effectiveness, and accuracy of your research.

Order this product and report back to us. Every feedback will be re warded!

Order No.: 50301

Using the mmi Live Cell Chamberon a microscope

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LEAD THE WAY to live cell microdissection

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LEAD THE WAY to an easy operation

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H Automatic documentationOptions

Toolbar and navigation overview

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Laser parameter controlClear and intuitive setting

INTUITIVE SOFTWARE The mmi CellTools software is used to control the mmi CellCut Plus system. Its graphical user interface allows the precise and intuitive identification and selection of the areas of interests to be excised for the isolation processes. In addition, it provides full control of the system, laser cutting parameters, type of objective used and the specific camera settings related to each different application.

With a fully motorized microscope as its basis, even the automated microscope functions such as objective changes, condenser settings and fluorescence turret changes can be initiated via the mmi CellTools software. With this user-friendly software combined with the optional PenScreen operation, the mmi CellCut system simply makes the most complex experiments possible and will take you to new levels of efficiency.

Overview, easy navigation and positive target identification

Using a low-magnification objective, an overview scan of the entire microscope slide is taken, allowing easy navigation of the whole slide. As well as to find areas of interests or positions, the mmi IsolationCap is then place in the inspection mode by clicking the mouse button. In addition, when using the mmi MultiSlide option, the overview and easy navigation are extended on all slides.

The user-defined inspection mode position is easily set by pressing a button on the naviga-tor screen and can be reset at any time.

Laser control

The common laser parameters such as speed, focus and power can be set to match the individual needs of each application, sample or objective. Once set, the parameters are stored for easy retrieval and can be edited at any time.

Automated full-slide selection and extraction

While the areas of interests are all over the entire microscope slide, you can select and extract from the whole slide area; whereas the software will automatically move the mmi CapLift over the slide to the right position and pick up the microdissected areas.This saves a significant amount of time in the isolating process. Together with the mmi MultiSlide and mmi MultiCap options, the system automatically extracts targets from up to three slides and collects in up to eight different mmi IsolationCaps.

Auto documentation

All the relevant data within a session can be stored via the auto documentation mode. This includes the number and size of the cut-out areas, as well as instrument settings and laser parameters. The isolated targets are documented by the mmi CellCut Plus system during the cutting and isolation process and pictures of the remaining samples are also au-tomatically taken and saved.This enhances traceability of your research work and a given proof of a clear, clean and quick isolation processes. Images can be saved as TIFF, JPEG, BMP or TGA files.

Key customer benefits:

• Easy to use and intuitive software interface enabling intensive focus on your research work

• All control parameters are clearly structured on display• Versatile plug-and-play options for individual needs• Simple and efficient auto documentation of your work• Same user-friendly interface for others MMI products for simple expan-

sions and upgrades

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SYSTEM INTEGRATIONInverted microscopes are designed to provide researchers with a high performance and versatility required for a wide range of research activities. The mmi CellCut Plus is fully integrated with the inverted microscope, maintaining all the properties and op-tions of these research instruments without experiencing any limitations while using our microdissection system. MMI has a fully integrated automatic microscope giving you all the advantages of these technological advances for direct control, easily and intuitively with our mmi CellTools software.

Optical bench flexibility

With multiple ports and dual-level laser integration, the microscope offers the “optical bench flexibility” enabling the use of different modules and imaging technologies to be used without any limitation or extensive equipment changes. The system-integrated digital camera is connected to the standard left side port; whereas, other cameras may be added at different ports according to your individual needs.

Standard fluorescence without limits

The dual-level coupling of the solid state UV-cutting laser also ensures that the microscope may be used fully for the whole range of fluorescence applications. For exa-mple, six fluorescence cubes can be used and therefore, additional or specialized dichotic optics or external filter wheels are not necessary.

In conjunction to the mmi CellTools software’s freeze mode for fluorescence images, the mmi CellCut Plus system provides a perfect combination for fluorescence applications and laser microdissection adaptable to your needs.

The L-shaped fluorescence condenser is attached for easy handling of the 100W mercury lamp house. Furthermore, all available options including the fibre optic cold fluorescence illumination or other external lamp houses can also be used.

The mmi CellCameras

MMI offers an ultra high sensitivity camera for laser microdissection applications. The new digital mmi CellCamera with its compact dimensions and outstanding performance supports your most complex demands.

• CellCamera DXA285cF for outstanding image quality and colour fidelity• IEEE 1394a (FireWireTM progressive scan CCD camera)• Ultra high sensitivity Exview CCD technology and dynamic range• Advanced passive cooling concept for exposure times up to 30 sec• 1392 x 1040 pixels• High Quality 12 Bit slow scan mode for lowest readout noise• Up to 15 full frames per second• Ultra compact dimensions• Automatic shutter, automatic/manual white balance

Key customer benefits:

• No laser interference on microscope functions due to dual level coupling• All standard microscope options remain intact while using the mmi

CellCut Plus• Freeze mode for fluorescence images avoids bleaching of cells• High quality imaging and contrast with mmi CellCamera• Cold fluorescence light source has 10x longer lifespan as regular lamp

with alignment free operation and no heating of the samples

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A Dual-level laser coupling Scheme of different light parts

Fluorescence up to six fluorescence filter cubes

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mmi CellCamera high resolution digital camera

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LEAD THE WAY to the future

D PenScreenfor on-screen target identification

E mmi CellExplorer software

mmi MembraneSlide

mmi IsolationCap

mmi CellChamber

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MODULAR UPGRADES AND OPTIONSFor more advanced functions and different requirements, the mmi CellCut Plus system can be upgraded and complemented with a number of useful additions.

PenScreen – A sensitive PenScreen can easily be used to operate the mmi CellCut Plus system in selecting areas of interests directly on the system display.

mmi CellExplorer – Image analysis software which automatically identifies cells based on selection criteria distinctly specified by morphological parameters predefined individually as such in size, shape, staining and fluorescent markers to allow for automatic dissection.

mmi MultiCap – allows automatic collection of targets in up to eight different IsolationCaps keeping the different targets segregated (e.g. MultiGroup function) or to increase throughput together with the mmi MultiSlide.

mmi MultiSlide – For simultaneous microdissection of up to three slides and to increase throughput.

SERVICES AND CONSUMABLES MMI as a leading supplier of LCM instruments also provides a variety of services and appli-cation support. This support is offered throughout the whole value-added chain of sample staining, over microdissection to nucleic acid and protein isolation and analysis. MMI and its partners provide worldwide technical support, guidelines as well as answers to all your questions.

Other MMI services are on-demand end-user training either in-house or on-site, protocols for various applications, scientific publication support, service and maintenance contracts, etc.

MMI also offers on-site protocol development through our own applications specialists’ right in your laboratory with your own equipment and personnel providing you with solutions suited to your individual needs.

MMI Consumables

mmi MembraneSlidesAvailable in PET or PEN mounted on metal frames or plastic frames in RNAse free quality

mmi IsolationCapAvailable in different sizes for specific applications types such as fluorescence or bright field imaging.

mmi CellChamberCell chamber with membrane and special Petri dish for live cell cultures

mmi µ-Slide 18-well18 well for high throughput live cell application. Live cell isolation with mmi IsolationCap.

Director SlidesA new Generation of Laser Microdissection Slides for a faster, simpler Proteomic Analysis. The DirectorTM slides are non-contact laser Microdissection slides utilizing a thin energy transfer layer bonded to a glass support. Applying a Laser Induced Forward Transfer (LIFT) technique, the photonic laser energy is converted to kinetic energy upon striking the energy transfer layer. The layer itself is vaporized and the selected feature instantly propelled into the collection tube.

First Choice in LMD RNA friendly basic Staining KitFast H&E Staining: the whole procedure is done in less than 5 minutes.

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10 LEAD THE WAY to a new level of microdissection

Newly designed mmi MultiCap Lift

mmi LaserBox

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New digital mmi CellCamera

mmi CellCut Plus

LEAD THE WAY TO A NEW LEVEL OF LASER MICRODISSECTION

A The third generation of a much acclaimed cutting-edge tool, mmi CellCut Plus combines several proven leading-edge technologies which enable laser microdiss-ection of paraffin and cryo samples, smears and even cell cultures fast and precise. The advancement of the mmi UV-Cut software allows greater capabilities in a user-friendly design.

mmi CellCut Plus: All the solutions you need!Choose mmi CellCut Plus in order to benefit from a range of new advantages that will simplify your work more effectively.

Higher level of automation

B New Caplift design: Newly designed single or multi mmi CapLift in a plug and play module with adjustable cap contact pressure. The complete CapLift can be removed giving full access to the microdissection stage for use in a regular motorized inverted microscope, optical tweezer or other micromanipulation studies.

Available Options:

To increase throughput, mmi MultiCap enables the use of eight mmi IsolationCaps which can be simultaneously attached, facilitating automatic switching from one cap onto the other during group cutting.

The MultiSlide functionality allows the simultaneous microdissection of up to three mmi MembraneSlides.

mmi CellExplorer colour recognition software which automatically recognizes cells under fluorescence and bright field.

Higher level of flexibility

C A more powerful solid state laser is available for application that demand higher lasing energy (mini gels, leaves, wood, thick mammalian tissues, etc.)

Digital camera

D New digital camera. The mmi CellCamera is an ultra sensitive camera with progressive scan possibilities. This will enable users to obtain a much greater image quality and contrast of their samples. A b/w version is also available for sensitive fluorescence applications.

Key Customer benefits:

• Easy operation with high level of automation• Outstanding image quality with the new digital mmi CellCamera• Safe reliable operation• mmi CellCut Plus is compatible to the Olympus IX71 / IX81 or Nikon

Ti-E / Ti-S inverted microscopes• Newly designed single or multi mmi CapLift in a plug and play module

with adjustable cap contact pressure• Ability to use your system with a motorized inverted microscope, optical

tweezer or other micromanipulation studies

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mmi CellCut PlusA

LEAD THE WAY to an easy micromanipulation

mmi CellManipulatorup to 10 independent trapping beams

mmi CellManipulator fully integrated with mmi CellCut Plus

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mmi CellManipulator softwareeasy to use software

A NEW GENERATION OF PROFESSIONAL OPTICAL MULTIBEAM TWEEZERSmmi CellManipulator is the world’s only available commercial optical multibeam tweezer system. This optical trap enables ultra-precise, contact-free manipulation of microscopic particles and the measurement of intracellular activities.

It can be used to study cell-cell interactions, in vitro fertilization or cell fusion. Cell sorting and cell positioning can also be accomplished together with an optional exclusive mmi 4-Quadrant detector enabling the measurement of binding forces or viscosities at sub cellular level.

Function

E The mmi CellManipulator uses a ND-YAG infrared laser with a long wavelength of 1064 nm with up to 3 W of trapping power. The wavelength of the laser does not interfere with the integrity of the living specimens. The intensely focused laser beam can hold, move, rotate, join, separate, stretch or otherwise manipulate up to ten microscopic objects simultaneously in three dimensions. The size of the particles can range from 0.1 to 200 micros in liquids. (System complies with IEC608225-1 Am. 2:2001: Class 1 laser product)

High-performance software

F Advanced software enables comfortable, contact-free, high-precision manipulation of se-lected particles inside the entire field of view. Each beam can be moved separately, and allows the user to define groups of tweezers points, which can be pushed, rotated, contracted and expanded, as groups or individually as required.

Modularity

G mmi CellManipulator can be used as a stand alone unit or it can be combined with the mmi CellCut Plus at any time to expand the research possibilities. This, for example, would allow the disruption of cells by permitting the introduction of structures or active substances or to initiate cell fusion or work on fluorescence options.

Options

For an even easier and more effective operation, customize or upgrade mmi CellManipulator with a choice of value-adding options such as: high precision 4-Quadrant detectors – use to measure binding forces and viscosities at sub cellular levels.

Key Customer benefits:

• Unique professional standards in expanding the frontiers of research• Comfortable and safe multibeam operation of up to 10 tweezers points• High performance laser which ensures the integrity of the living specimens• Contact-free and precise movement of selected particles individually or in

groups• Easy operation with live video and documentation

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mmi CellManipulator

mmi CellCut Plus SPECIFICATIONSItem Specifications

mmi CellCut Plus can be delivered together with Nikon Ti-E / Ti-S series or Olympus IX71 / IX81 series.

MMI Inc.P.O. Box 348Haslett, MI 48840, USAPhone: +1 603 629 9536Fax: +1 321 978 0304sales_us@molecular-machines.comwww.molecular-machines.com

Samples For all relevant application samples (e.g. cryo or paraffin-preserved tissue,single cells, cell compartiments, cytospins, chromosomes etc.)

"Live Cell" handling Positive and negative cell selection / isolation possiblePicosecond UV, solid state laser Computer controlled

Wavelenght: 355 nmPulse duration: <500 psecPulse energy/average energy: <1 joule/appr. 4 mWRepetition rate: <5 kHzSystem complies with IEC 60825-1 Am. 2: 2001. Class 1M laser product (invisible laser radiation. Do not view directly with optical instruments)

CapLift technology SW-controlled, covering full slides; unique and contamination free technologyDigital camera with ultra high sensivity Digital colour: 2,088 x 1,550 pixels or

Digital colour: 1,392 x 1,040 pixels orDigital monochrome: 1,392 x 1,040 pixelsCompact housing and FireWire connection

UV-Cut software basic functions Laser energy and focus controlFull slide and Petri dish controlInspection mode with positive target identificationSaving multi-user profilesMultiGroup functions across entire samples / slidesAutodocumentation for sample, images and parameters

PC and Monitor Specifications will be continuously updated according to market developmentsWindows XP Professional, 19" LCD Monitor (with high contrast)

Motorised stage Computer-controlled for high-precise movement/cuttingTravelling range: 112x74 mmStep widht: 0.075 mRepositioning accuracy: 1 m

OptionsPen Screen system operation Sensitive 17" or 21" touch screen monitor for user friendly system operation

and to allow direct target identification with a special penmmi CellExplorer image analysis software Identifies and cuts automatically defined targets based on user settingsmmi MultiCap (motorized) Allows automatic collection of targets in up to 8 different IsolationCapsmmi MultiSlide For microdissection of 3 slide assemblies

X-Cite metal halide illuminator cold light, no adjustments, 10x longer lifespan

Possible CellCut system upgrademmi CellManipulator optical tweezer system Ultra precise, contact-free manipulation of microscopic particles,

with up to 10 independent beams on a high-quality, Nd: YAG infrared laser

Laser Microdissection System

mmi SmartCut Plus

Molecular Machines & Industries

IntroduCtIon

At the CuttIng edgethe mmi SmartCut Plus system consists of a comfortable inverted microscope with a motorized scanning stage, a solid state laser with a requisite laser beam delivery, transfer optics and a high-end computer which runs under MS Windows. the system is operated via MMI’s especially developed software mmi Celltools, which controls the laser, image capture, all scanning stage actions and automated mmi CapLift options.

A In microdissection a “live view” image of a microscopic sample is displayed on the monitor, allowing the user to easily identify and mark the target area e.g. a cell or a group of cells. during microdissection, the uV laser is focused by the objective onto the microscopic sample and remains aligned at the centre of the optical axis whilst the high-precision motorized stage is used to accurately move the sample.

high speed and ultra fine laser microdissection is achieved via

A maintenance free uV solid state laser üoptical laser focusing to a cutting line < 1 µm üShort laser pulse duration in picoseconds and high repetition rate of >5 khz ü

Furthermore the very low laser pulse energy (<1 µjoule), the mmi SmartCut’s Plus “cold ablation” process leaves the target material unaffected with no impact on subsequent protein analysis, dnA or rnA extraction. even live cells can be isolated and cultured after microdisscetion. And the unique and patented mmi CapLift technology provides the contamination-free and targeted collection of microdissected sample material.

the mmi SmartCut Plus is easy and convenient to handle, requires no extensive tech-nical training and does even fit under a laminar flow hood. the mmi SmartCut Plus is especially developed for all routine applications and researchers in cell biology, molecu-lar pathology, forensic medicine prenatal diagnostics and many other fields will greatly benefit from it’s unique features.

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Principle of microdissectionFast, precise and contamination-free

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Computer-controlled movement

Celltools software-controlled laser focus and energy

LeAd the WAY For LCM reduCed to the MAxnature provides us with a huge variety of precise and fast dissection systems. For example the Leaf Cutter Bee - the first bee seen in early spring. She darts to a suitable tree, and within two minutes, selects a leaf, cuts a hole out, and then quietly departs.

MMI Switzerland developed and optimized solid state laser technology for the ultra-precise cutting of samples from tissue and live cell cultures. today, this technology is emerging as a prime technology for the rapid isolation of single cells or multicellular structures.

hereby, we introduce our latest & smartest innovation: the mmi SmartCut Plus system - a compact and high-precision laser capture microdissection system (LCM) that will impress with it's fast and easy handling and not least with it's cost effectiveness.

LeAd the WAY to cutting edge technology

SAMPLe PrePArAtIon CuttIng & reCoVerY

Unique contamination-free sample preparationB MMI's uniques sample preparation technique effectively protects the sample by form-ing a complete barrier to contaminants, such as environmental impurities, which is essen-tial for laser microdissection steps. to isolate targets from any source, including cryo or paraffin-preserved tissues, smears and cytospins, the mmi SmartCut Plus uses a metal frame slide with a 1.4 µm Pet-membrane. the different types of samples are placed directly on the membrane, which is then inverted and a standard glass slide placed below. In order to optimise the preparation of delicate samples, standard protocols – such as the use of poly-L-lysine coating or uV exposure to increase cell adhesion – can still be used. thus, the sample is ready for the subsequent, highly pure isolation via the adhesive cap of the mmi IsolationCap.

Selection and cuttingC targets of interest to be extracted are selected on-screen using either free-hand or predefined geometric shapes, such as circles, squares and ellipses. Any number of areas across the entire section can be identified and the sizes of the geometric shapes can be changed as well as copied and pasted for consistency. A group function allows the user to collect an unlimited number of different cells or cell compartments within one screening process in different microtube caps the <1 µm cutting line enables the precise excision of the selected targets at an out-standing speed, without affecting the target. As a result, there is no loss in quality of the material used downstream. even the viability of living cells is not affected and therefore they can be re-cultured once selected.

With such fine control over cutting, mmi SmartCut Plus can be used on a wide range of sources, making it a highly flexible tool for most laboratories. target cells are extrac-ted without any damage to their proteins, dnA or- rnA, and as a result, downstream analyses can be carried out with total confidence.

Target recovery with the automatic mmi CapLift technologyd the unique mmi CapLift technology has been developed to maintain the ease of use and contamination-free nature of the mmi SmartCut Plus system. Sample uptake after microdissection is carried out automatically with the newly designed single mmi CapLift technology. the mmi IsolationCap is centred to the optical axis of the system to enable user-friendly collection of the selected targets. With the cap kept centred in the optical axis, the motorised stage is moved to enable collection of target cells from across the entire slide. this plug in module does not only allow for a targeted and traceable sample uptake of dissected samples it even allows to adjust the cap contact pressure for sample uptake.

Isolated targets, placed on the cap of the mmi IsolationCap, are ready for subsequent downstream processes such as dnA and rnA extraction, as well as protein analysis. Isolation buffer is added and the tube closed for centrifugation to keep the recovery process as short and delicate as possible. this unique feature ensures full control of the work flow and thus provides safety and reliability for tricky application e.g. single cell analysis

A Lens offset function ensures convenient orientation on the slide after changing the objective as the selected area stays centered.

Live cell culture microdissection in our mmi CellChamberse using a mmi CellChamber, the mmi SmartCut Plus can be used to dissect living cells under sterile conditions. the cells grow in the chamber on a membrane coated with poly-L-lysine and, prior to microdissection, are placed in a Petri dish with an adhesive base. Both parts can be sterilised. After microdissection, a different recovery method is used. here the culture chamber is removed from the Petri dish under aseptic condi-tions, leaving the isolated cells behind.

Sample preparationon frame slide with membraneB

excision of live cellse

Special cell chamber with membrane

Petri dish with adhesive material

automatic mmi CapLiftd

easy cell selectionC

LeAd the WAY to an easy operation

mmi SMArtCut PLuS oPtIonS

mmi customized featuresthe mmi SmartCut Plus can be customized with the following choice of features:

F A PenScreen, which enables the selection of target cells and components directly on a touch-sensitive screen using a special pen. this allows the user much greater accuracy in selection, especially for free-form shapes.

g the mmi SmartCut Plus is also available with the mmi Cellexplorer , a cell identification software based on colour recognition, which automatically identifies and cuts-out cells,

h Some application may require the work with our manually adjustable mmi CapLift. Isolated targets, placed on the cap of the microtube, are ready for subsequent down-stream processes such as dnA and rnA extraction, as well as protein analysis. Isolation buffer is added and the tube closed for centrifugation to keep the recovery process as short and delicate as possible.

IntuItIVe ControLthe mmi Celltools software is used to control the mmi SmartCut Plus system. Its graphical user interface allows precise and intuitive identification of the target areas to be excised for the subsequent isolation processes. It also provides full control over the laser cutting parameters, the objective used and specific camera settings in respect of different application-specific requirements. Moreover, all system functions are struc-tured in a way to make them self-explanatory, providing easy operation without any sophisticated technical knowledge or training. Some examples are given below.

Overview and easy navigation I using the 4x objective, an overview scan of the entire microscope slide can be

made, allowing the user to operate and navigate the system more easily

Cutting laser control to maximise the laser microdissection process, all laser parameters, such as speed, focus and energy, can be changed to match the individual needs of the application, sample and objective. once set, the parameters are stored for each process and can be easily recalled for use or editing

AutodocumentationAll the data relevant to a session are stored via the autodocumentation mode. this includes the number and size of the cut-out areas, as well as instrument set-tings and parameters. the isolated targets are documented by the mmi SmartCut Plus system during the cutting and isolation process, and pictures of the remain-ing sample are automatically acquired and saved – all with multi-user access. this enhances traceability and is proof of a clear, clean and quick isolation process.

I toolbar and navigation overview

PenScreenFor on-screen target identification

F

manual mmi CapLifth

g mmi CellexplorerFor cell identification

ChAPter I

one tooL: VArIouS APPLICAtIonSmmi SmartCut Plus is an extremely versatile instrument, across different applications. Whether you are isolating specific cells from an entire tissue, maintaining a pluripotent stem cell line, or extracting a telling foreign cell for forensic investigation, mmi SmartCut performs perfectly.

Pathological, haematological and cytological samplesJ today, targeting specific cells in healthy as well as in diseased tissues is of particu-

lar importance in elucidating the molecular mechanisms that lead to cancer and other life threatening diseases. the group of tomlins et al. published recently a molecular concept of prostate cancer progression after laser microdissection and individual profil-ing of 101 specific cell populations with the mmi CellCut, (ref.1), further Buckanovich et al. used Immuno-LCM for the expression profiling of cell populations in the tumor microenvironment of human ovarian cancer (ref.2.) Figure J showed a colon tissue section stained with hematoxylin/eosin where the insert displays intestinal glands from this section after microdissection. the easily dissected area is now ready for any down-stream analysis such as molecular profiling for proteomics or genomics means. ref. 1 tomlins, S.A. et al. nAture genetICS 39, 41-48 (2007)

ref. 2 Buckanovich, rJ et al. CAnCer BIoLogY & therAPY, 5:6 635-642 (2006)

Forensic investigationsK the mmi SmartCut Plus system is an extremely useful tool in forensic medicine, since it can be used, for example, to isolate a single sperm from a vaginal smear for genetic analysis, leading to traceable dnA fingerprints. In a recent work published by Anslinger et al. the sex-specific isolation of cells from mixtures by means of combined in-situ hybridization and single cell isolation via laser microdissection has been judged as a tool that will greatly facilitate forensic work. ref. 3 Anslinger, K. et al. Int J Leg Med 121 (1), 54-56 (2007)

Live cellsL Most stem cell lines are presently grown at high densities on mouse fibroblast “feed-er layers”. therefore, isolating specific pluripotent cells from the surrounding fibroblasts and differentiating cells needs to be rapid, exact and easy. mmi SmartCut Plus offers you the perfect balance between speed, precision and ease of use.

J Intestinal gland isolated from transverse tis-sue sections of colon

Single sperm isolated from a gynaecological smearK

LeAd the WAY to a wide range of applications

Cell culture in phase contrast observationL

the manufacturer reserves the right to make technical changes without prior notice.

mmi SmartCut Plus SPeCIFICAtIon

Item Specifications

Samples For all application-relevant samples; cryo or paraffin-preserved tissues, single cells, cytospins, cell compartiments, chromosomes, etc.

Microscope System integrated nikon tS100-F with CF160 infinity optics, illumination pillar with 30 W halogen lamphouse, long working distance condenser nA 0.3 Wd = 75mm,

CF160 objectives a) CFI PLan Fluor objective with 4x magnification, nA 0.13 and Wd of 17.1b) CFI PLan Fluor eLWd objective with 20x magnification, nA 0.45, Wd from 7.0 to 8.1

Picosecond uV, solid-state laser Computer-controlled; Wavelength: 355 nmPulse duration: <500 psecPulse energy/average energy: <1 µjoule/approx. 4 mW repetition rate: > 5 khzSystem complies with IeC 60825-1 Am. 2: 2001. Class 1M laser product(invisible laser radiation. do not view directly with optical instrument)

CapLift technology Covering full slides; unique and contamination-free sandwich technologydigital camera with high sensitivity digital colour: 1032 x 776 pixels with integrated 1/3” interline progressive CCd

Compact housing and FireWire connectionmmi Celltools software basic functions Laser energy and focus control

Full slide and Petri dish controlSaving multi-user profilesMultigroup function across entire sample/slidesAutodocumentation for sample, images and parameters

PC and monitor Windows xP, 19” LCd monitorSpecifications will be continuously updated according to market development

Motorised stage Computer-controlled for high-precision movement/cuttingtravelling range: 120 x 100 mm; step width: 0.075 µm; repositioning accuracy: 2 µm

OptionsPenScreen system operation Sensitive 17” or 21" touch screen monitor for user-friendly system operation

and to allow direct target identification with a special penmmi Cellexplorer A cell identification software based on colour recognition which automatically identifies

and cut's out cellsmanual mmi CapLift manual mmi CapLift available upon request

MMI Inc.

P.O. Box 560728

Rockledge, FL 32956-0728

Phone: +1 603 629 9536

Fax: +1 321 978 0304

Mail : sales_us@molecular-machines.com

www.molecular-machines.com

MMI GmbH

Breslauer Strasse 2

85386 Eching, Germany

Phone : +49 89 319 048 40

Fax : +49 89 319 048 59

Mail : info@molecular-machines.com

www.molecular-machines.com

MMI AG

Flughofstrasse 37

8152 Glattbrugg, Switzerland

Phone: +41 44 809 10 10

Fax: +41 44 809 10 11

Mail: info@molecular-machines.com

www.molecular-machines.com

mmi SmartCut Plus can be delivered optionally with a nikon tS100-F or a olympus CKx 41 microscope.

Swiss Precision