2
‡ Simple operation and utmost reliability‡ Ultrafine sensitivity through high-precision pressure setting ‡ Maintenance-free piston/cylinder system‡ Large, ergonomic dials ‡ Universal capillary holder for mounting onto all
common micromanipulators
Intracytoplasmic sperm injection (ICSI)
Eppendorf Systems ...
CellTramCellTram® Air, CellTram® Oil and CellTram® vario— manual microinjectors for pressure control, manual microinjection, and liquid dispensing—are designed with special emphasis on optimal ergonomics, operational comfort, and high precision. CellTram microinjectors are the ideal complement to your Eppendorf workstation, yet they also fit into all other common micromanipulator workstations.
Intracytoplasmic sperm injection (ICSI), the injection of a single sperm into an oocyte, has become a powerful means of overcoming male factor infertility. For this application, Eppendorf offers the TransferMan® NK 2 micromanipulator. One manipulator is used for moving the holding capillary and a second for transferring the sperm. The oocyte is held in place using the manual injector CellTram® Air. Sperm transfer is carried out using the hydraulic manual injector CellTram® vario.
CellTram specifications CellTram Air CellTram Oil CellTram vario
Method of pressure generation Cylinder/piston system, air-filled, maintenance-free
Cylinder/piston system with QuickValve-System, oil-filled, maintenance-free
Cylinder/piston system with gear and QuickValve System, oil-filled, maintenance-free
Volume alteration per revolution 88 µl 9.5 µl 9.5 µl/950 nl (coarse/fine)
Selectable total volume 2,640 µl 950 µl
Total piston stroke 15 mm (corresponds to 30 revolutions)
50 mm (corresponds to 100 revolutions)
Minimum adjustable volume <200 nl <20 nl < 20 nl/< 2 nl (coarse/fine)Maximum pressure 2,900 hPa 20,000 hPa
Additional Leica Microscope Setup for IMSI:
Objective 100x oil
Contrast options DIC
Other requirements MagChange 1.6x, 2.0x, digital camera with C-mount Vario-TV-Adapter 0.33x–1.6x
Leica Microscope Setup
Microscope Inverted
Objectives Bright field 5x/10x, L 20–40x(Corr)
Contrast options IMC (Integrated Modulation Contrast)
Other requirements Stage with insert for petri dishes, heated stage or insert
recommended microscopes: Leica DMI3000 B - Leica DMI6000 B
Benefits
2
3
Embryonic stem cell transfer
Eppendorf TransferMan NK 2This micromanipulator is especially designed for work with suspension cells. It combines simplicity and innovation in a user-friendly system that satisfi es highest sophisticated user requirements.
With the smooth, stepfree proportional kinetics and menu-driven control, TransferMan NK 2 offers an intuitive operation, allowing the most complex micromanipulation techniques to be carried out rapidly, easily and precisely.
... plus qualifi ed Microscopes ...
Embryonic stem cell transfer is used to insert, as well as to modify DNA sequences of the animal genome.
For the transfer of ES cells into blastocysts, two TransferMan NK 2 devices are used for controlling the holding and transfer capillary. CellTram Air holds the blastocyst while the CellTram vario is used for transferring the ES cells.
TransferMan specifi cations
Control Via central joystick; proportional kinetics
Drive Stepper motor
Resolution Approx. 40 nm per micro step
Max. movement (each axis)
Approx. 20 mm
Max. speed of tool 7,500 µm/s
‡ One single joystick for coarse/fi ne movements and control of all axes
‡ Proportional kinetics provide intuitive control‡ Automated Home function for fast capillary exchange‡ Individual speed setting allows to work at full capacity‡ Storage of three independent work positions
Benefi ts
Leica Microscope Setup
Microscope Inverted
Objectives Bright fi eld 5x/10x, L 20–40x
Contrast options IMC (Integrated Modulation Contrast), DIC
Other requirements Stage with insert for petri dishes, heated stage or insert
recommended microscopes: Leica DMI3000 B - Leica DMI6000 B
3
4
Pronucleus injection
FemtoJetThis microinjector features a wide range of functionality, simple operation and a built-in compressor to indepen-dently deliver the required pressure. It is ideal for injecting aqueous solutions into adherent cells with InjectMan® NI 2 and suspension cells (with TransferMan NK 2). The FemtoJet forms the perfect system with Eppendorf micromanipulators, and it integrates just as easily with non-Eppendorf microinjection systems.
... result in excellent Workplaces
The pronuclear microinjection method of producing a transgenic animal is based on the introduction of linear DNA sequences into the chromosomes of fertilised eggs.
The TransferMan NK 2 micromanipulator in combination with Leica microscopes, FemtoJet® and CellTram Air forms an ideal system to satisfy high demands on accuracy and functionality.
FemtoJet specifi cations
Pressure supply Integrated compressor
Injection time (Ti) 0.0–99.9 s; can be set in increments of 0.1 s
Compensation pressure (Pc)
0; 5 hPa-max. 6,000 hPa (87 psi); can be set in increments of 1 hPa
Rinsing pressure Max. 7,000 hPa (102 psi)
(W × H × D) 22 × 28 × 17 cm
Weight 6.2 kg
‡ Independent pressure supply with built-in compressor‡ Programmable injection ensures reproducibility‡ Inject and Clean functions triggered via control button
on instrument, supplied mouse or optional foot control‡ Electronic linkage to InjectMan NI 2
Benefi ts
Leica Microscope Setup
Microscope Inverted
Objectives Bright fi eld 5x/10x, L 20–40x
Contrast options IMC (Integrated Modulation Contrast), DIC
Other requirements Optional fl uorescence, temperable stage or insert
recommended microscopes: Leica DMI3000 B - Leica DMI6000 B
4
5
... meet Microscopes
InjectMan NI 2This ultra-precise micromanipulator was designed in response to researchers' suggestions and application requirements. As a result, the InjectMan NI 2 is your ideal partner for microinjection into adherent cells as well as for microinjection into early stage embryos and small organisms.The unique ability to connect the InjectMan NI 2 with our microinjectors FemtoJet and FemtoJet express ensures a very effi cient and reproducible processing.
Microinjection is one of the core methods to introduce foreign DNA and other non-permeable molecules into cells. Nuclear injection of plasmid DNA enables rapid expression of proteins in specifi c cells within a population.
The menu-controlled, programmable micromanipulator InjectMan NI 2 is ideally suited for microinjection of adherent cells. The electronic connection of the InjectMan NI 2 and the FemtoJet guarantees a very rapid, safe and reproducible microinjection process.
InjectMan specifi cations
Control Via central joystick; dynamical kinetics
Drive Stepper motor
Resolution Approx. 40 nm per micro step
Max. movement (each axis)
Approx. 20 mm
Angle of inclination 30–60° to object table
Max. speed of tool 7,500 µm/s
‡ Precise coarse/fi ne control of all axes with one single joystick‡ Axial mounting of the Z-module‡ Can be used with FemtoJet or FemtoJet express for
automated axial injection movement‡ Automated Home function for rapid capillary exchange‡ Programmable Z-axis limit for defi ned injection level
avoids capillary breakage
Benefi ts
Leica Microscope Setup
Microscope Inverted
Objectives 10x, L 20–40x
Contrast PH (Phase contrast), IPH (Integrated PH)
Other requirements Fluorescence, stage with insert for petri dishes
recommended microscopes: Leica DMI3000 B - Leica DMI6000 B
5
Microinjection into adherent cells
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Your local distributor: www.eppendorf.com/worldwideEppendorf AG · 22331 Hamburg · Germany · Tel: +49 40 538 01-0 · Fax: +49 40 538 01-556 · E-mail: [email protected]
Eppendorf North America, Inc. · One Cantiague Road · P. O. Box 1019 · Westbury, N.Y. 11590-0207 · USA Tel: +1 516 334 7500 · Toll free phone: +1 800 645 3050 · Fax: +1 516 334 7506 · E-mail: [email protected]
Application Support Europe, International: Tel: +49 1803 666 789 · E-mail: [email protected] America: Tel: +1 800 645 3050 ext. 2258 · E-mail: [email protected]
Asia Pacific: Tel: +60 3 8023 6869 · E-mail: [email protected]
Ordering information
Description Order no.
Micromanipulator InjectMan® NI 2
manipulator for semiautomatic microinjection into adherent cells 5181 000.017
TransferMan® NK 2
micromanipulator for suspension cells with menu-controlled programming 5188 000.01210° adapter 5188 110.004A-head1 for 10° assembly 5181 150.027
Microscope adapter
Leica DM IL LED 5181 212.006 Leica DMI3000 B/4000 B/6000 B 5181 203.007Universal Stand2, suitable for mounting of manipulators on upright microscopes and stereo microscopes independent of the microscope tripod used
5181 250.005
Microinjectors
FemtoJet®, programmable microinjector with integrated pressure supply 5247 000.013FemtoJet® express, programmable microinjector with external pressure supply 5248 000.017
CellTram
CellTram® Air, pneumatic manual microinjector 5176 000.017CellTram® Oil, hydraulic manual microinjector 5176 000.025CellTram® vario, hydraulic manual microinjector, with gears 5176 000.033
Consumables
Femtotip®, sterile injection capillaries, 0.5 µm inner and 1.0 µm outer diameter, set of 20 5242 952.008Femtotip® II, sterile injection capillaries, 0.5 µm inner and 0.7 µm outer diameter, set of 20 5242 957.000Microloader, 2 racks of 96 5242 956.003VacuTip, sterile microcapillaries for holding suspension cells; 35° angle; 15 µm inner diameter, 100 µm outer diameter (set of 25) 5175 108.000TransferTip®-RP (ICSI), sterile microcapillaries for sperm injection with the ICSI technique; rigid, parallel flange; 0.5 mm long; 35° angle; 4 µm inner diameter, 7 µm outer diameter (set of 25)
5175 114.000
TransferTip®-F (ICSI), sterile microcapillaries for sperm injection with the ICSI flexible flange; 0.4 mm long; 35° angle 4 µm inner diameter, 7 µm outer diameter (set of 25)
5175 106.008
TransferTip®-R (ICSI), sterile microcapillaries for sperm injection with the ICSI rigid flange; 1 mm long; 35° angle; 4 µm inner diameter, 7 µm outer diameter (set of 25)
5175 113.004
TransferTip® (ES), sterile microcapillaries for transferring ES cells into blastocysts; 20° angle; 15 µm inner diameter, 20 µm outer diameter (set of 25)
5175 107.004
1 The assembly of the 10° adapters, together with the A-head, enables work with very shallow injection angles (e.g., for injection into the pronucleus of mouse embryos). 2 The universal stand is suitable for the assembly of manipulators independent of the microscope tripod used.
For more information, visit our webpages at www.eppendorf.com/cellmanipulationand www.leica-microsystems.com