CellSurgeon

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Subcellular Laser Dissection with Nanometer Precision

The CellSurgeon is a femtosecond laser based nanodissection system, designed for gentle and contact-free manipulation of cells and subcellular structures.  It enables very precise surgery inside live cells, without causing cell damage.

Overview

The CellSurgeon is a microscope based laser dissection system. CellSurgeon is the right instrument to impact cell dynamics, to manipulate cell organelles. Furthermore, cellular and subcellular manipulations in live cells and subsequent high resolution multiphoton imaging can be done easily with CellSurgeon. 

  • Contact-free nanodissection of cells and subcellular structures
  • Gentle manipulation in live cells, without loss of vitality
  • High precision and reproducibility
  • CellSurgeon module is adaptable to various microscopes and imaging procedures
  • Easy-to-use software for analysis, image processing and automated procedures
  • Cost-effective retrofitting of existing imaging systems
  • Customized configuration

 

CellSurgeon enables precise, contact-free 3D-nanodissection of cells and subcellular structures. CellSurgeon can be used to influence cell dynamics, to deactivate cell organelles, or to influence metabolic processes. Feasibility of a wide range of different applications has been already shown. For example, optical perforation of cell plasma membrane to enable DNA uptake as well as cutting of cell structures such as cytoskeleton or manipulation and subsequent time-lapse imaging in 3D.

Main component of CellSurgeon is a pulsed near-infrared femtosecond laser. The laser is coupled into microscope and the beam is tightly focused by high numerical aperture objective. Inside the laser focus very high intensities occur, inducing nonlinear optical processes and finally leading to ablation. Cuts generated in that way are limited to dimensions within the range of nanometers and thus very precise. Due to virtually no thermal or mechanical energy transfer, the cellular structures adjacent to the tight focus of the laser beam remain unharmed and intact. This, especially, is of great benefit in case of manipulation of subcellular structures within live cells because the risk of cell death is insignificant low. However, manipulation processes conducted by CellSurgeon are not only of high precision but they are also highly reproducible.

The ultrafast near-infrared laser with which CellSurgeon is equipped can not only be used for dissection but also for true confocal imaging. CellSurgeon is a multiphoton microscope which allows you to generate high resolution confocal 3D-images of either stained or marker-free samples. Moreover, due to high penetration depth of near-infrared laser, CellSurgeon is the ideal tool to perform imaging as well as manipulation in thick samples.

Another advantage of CellSurgeon is its adaptability to various microscopes and imaging procedures. ROWIAK offers customized solutions for different applications. Easy-to-operate software of CellSurgeon includes modules for 3D-dissection, confocal image acquisition, and basic image processing.


Features and Benefits

Dissection - femtosecond near-infrared laser 750-1050 nm

  • Precise 3D-cutting
  • No sample preparation needed
  • Cutting in native samples
  • Subcellular cutting
  • Cutting in living cells or living small organisms

Imaging - Femtosecond near-infrared laser 750-1050 nm

  • Multiphoton confocal imaging
  • Deep penetration imaging
  • Marker-free imaging possible

Laser movement by optics

  • Wide range of objectives can be used
  • Use of high N.A. objectives results in higher cutting precision

User defined profiles for laser cutting

  • Short set-up time
  • Cutting profiles for different samples can be stored and recalled

Built in laser calibration

  • Easy to perform
  • Fast alignment of laser beam

Pre-defined cutting shapes

  • Easy and fast selection of every structure of interest

Draw on screen function

  • Exact selection of structures

Fully motorized cutting module

  • Easy to use
  • Reproducible set-up
  • Increased throughput

Simultaneous 3D cutting and multiphoton observation

  • Allows selection and documentation in multiphoton mode of structures to be cut
  • Cut and observe (Time Lapse Experiments)

One system for a wide range of fluorescence applications

  • No additional imaging system needed

Versatile and easy to use Windows based imaging & cutting software

  • Short training times
  • Multi user friendly

Integrated design

  • Optimal performance
  • Responsibility by one company

Worldwide service and support

  • Fast service and support
  • Software maintenance from remote
  • Maintenance-free laser

Active application support

  • Support on application development

The CellSurgeon provides a powerful technology platform for precise and gentle nanosurgery in live cells. It is a versatile tool suitable for wide range of applications in biological, medical and pharmaceutical research. The combination of multiphoton imaging techniques with laser nanodissection enables novel experimental approaches to study cellular processes such as metabolism, cell signaling, cell division or apoptosis as well as cell dynamics. Depending on laser pulse energies manipulation levels can vary from stimulation to remodeling or even to complete ablation of cell structures.

ZMTH3 cells, transfected with pEGFP-HMGA2 by optical perforation, R: fluorescence image, L: brightfield image, with courtesy of Judith Baumgart, LZH Hanover

One distinguished application is the optical perforation* of cell plasma membrane to enable cells to uptake DNA, RNA, or proteins. By focusing the laser beam for a few milliseconds close to the membrane, transient pores are created so that nucleic acids or proteins can enter into the cell. Compared to conventional transfection techniques as the use of viral vectors, chemical carriers or electroporation, optical perforation considerably increases the transfection efficiency, and decreases the risk of cell damage.


* CellSurgeon module for transfection of molecules into living cells is available only with an additional module for frequency doubling of the laser frequency. Please contact our product manager for further information.

Ablation of a Single Mitochondrium

From optical knockout of specific cell organelles to dissection of single cytoskeletal filaments - the CellSurgeon allows very precise and reproducible nanosurgical operations. Dissection and ablation of subcellular structures do not affect surrounding structures [Fig. 1].


 

CellSurgeon - For Chromosome Dissection and Extraction

CellSurgeon equipped with Micromanipulators and a capillary holder can now be applied for microdissection of single chromosomes [Fig. 2a] and extraction of chromosomal fragments [Fig. 2b].

The laser beam of the CellSurgeon can be used to dissect the chromosomes to fragments of less than one micron (approx. 60 Mb of DNA) in thickness and isolate a single fragment for subsequent processing such as sequencing, or cloning and construction of libraries.

Since we use near-infrared laser for microdissection, the quality of DNA adjacent to the cutting line is hardly affected by the long-waved light.

Fig. 1 Endothelial cells before (R) and after (L) ablation of a single mitochondrion, by courtesy of Judith Baumgart, Laser Center Hanover
Fig. 2a Dissected chromosomes
Fig. 2b Extraction of dissected fragment

The CellSurgeon is not only a nanodissection instrument. Depending on the laser energy it is also possible to use the system for bleaching of labeled cell structures. A further potential application is the optical stimulation of cell organelles.

GFP-labelled mitotic spindles (fission yeast Schizosaccharomyces pombe) L: bleaching, R: cutting/ ablation, with courtesy of Isabell Raabe, MPI of Cellular Biology & Genetics

Customized configuration

LLS ROWIAK LaserLabSolutions offers customized configurations. Depending on your requirements different lasers, positioning systems and microscopes are available. It is also possible to adapt the CellSurgeon to microscopes and imaging procedures already existing in your lab. Please contact us for more information!


Standard Equipment

SmartBeam
Laser light source

  • Coherent Chameleon
  • Tuning range 720-950 nm
  • Repetition rate 90 MHz
  • Output power 1 W

SmartDissectionPlus
Pulse picker + controller

flexible adjustment of the pulse repetition rate of the laser for extreme sensitive cutting studies


SmartDissection
Positioning controls

  • Motorized XY stage and joystick
  • Piezo objective z-drive 500 nm
  • Shutter and controller
  • Specimen holder for glass slides and petri dishes
  • PC and Monitor

SmartMicroscope
Inverted Microscope Stand

  • 12 V / 100 transmitted light illumination
  • Front, side, and bottom photo ports
  • Manual Z focusing drive
  • Binocular tube
  • 6x objective revolver
  • Long distance condenser 0,55 DIC, PH, 5x with aperture
  • HBO100 W incident light source
  • Incident light attenuator
  • 6x motorized and coded filter cube turret
  • Filter 01 BP 365 / 12 FT 395 LP 397
  • Filter 10 BP 450-490 FT 510 BP 515-565
  • Filter 31 BP 565/30 FT 585 BP 620/60

SmartObjective

  • Objective  20x / 0,50
  • Objective 40x / 1,3 Oil
  • Objective 100x/1,3 Oil

SmartImage

  • 0.63x front port c-mount adapter
  • Fluorescence digital camera

SmartImagePlus
Multiphoton imaging

  • Photomultiplier
  • Amplifier
  • External filter wheel
  • Filters: e.g.  short pass, DAPI, GFP, TexasRed

SmartLab
Optical table

  • ScienceDesk Heavy Duty Passive Frame 900 x 1200mm
  • PerformancePlus Breadboard - 1200 x 900 x 60mm

SmartSoftware

Core software for nanodissection and image acquisition

Product Brochures & Application Sheets

Related Material

Images

Contact

Dr. Fabian Will
Phone +49 (0)511 277 2953
Fax +49 (0)511 277 1766
f.will(at)lls-rowiak.de

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Belgium

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info(at)autoqbiosciences.com

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Romania

APEL LASER S.R.L.
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077135, Mogosoaia, ILFOV, Romania
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www.apellaser.ro
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ab ninolab
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Taiwan

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