Author Archives: Leslie Knöbel

Nano-Plotter – Piezoelectric Pipetting Tips

Piezoelectric Pipetting Tips

The Nano-PlotterTM piezoelectric pipetting tips consist of glass and silicon. They are achieved from microstructured glass/silicon wafers.

 

The tips tie into the instrument’s fluidic system through metal shafts. Each pipet connects to a dedicated syringe that supplies both sample and washing fluids. NP1.2 runs up to eight tips, all N2.x instruments can be equipped with 1…16 tips.

 

Tips for different drop sizes are available:

  • Nano-Tip J:     0.35…0.4 Nanoliter
  • Nano-Tip A-J: 0.25          Nanoliter
  • Pico-Tip J:       0.06          Nanoliter
  • Nano-Tip J-H: 0.35…0.4 Nanoliter (External heater unit required)
  • Pico-Tip J-H:   0.06          Nanoliter (External heater unit required)

 

Tips for particular applications:

  • Nano-Tip HV-J: For high-viscous samples
  • Nano-Tip AR-J: Slim design,for array printing into 96-well plates

 

The Nano-Plotter piezoelectric pipetting tips consist of glass and silicon.

Pipetting Tips

Non-contact Micro Dispensers work on any Substrate Surface

The Nano-PlotterTM piezoelectric pipetting tips for the Picolitre range consist of glass and silicon- they are valveless and fabricated from microstructured glass/silicon wafers. Alternatively the instrument is available with solenoid valve dispensers for the Nanolitre range.

The tips interface with the instrument’s fluidic system through metal shafts. Each pipet connects to a dedicated syringe that supplies both sample and washing fluids. All N2.x instruments can be equipped with 1-16 tips in any combination.

Due to the nature of non-contact dispensing the Nano-Plotter supports so called Spot-On-the-Fly mode (SOF, since 2017) for microarraying applications with only one drop per spot position. Particularly when processing a larger number of flat substrates (E.g. glass slides) SOF saves up to 50% time.

Piezoelectric Tips for the Picolitre range

  • Nano-Tip J:     0.35…0.4 Nanoliter
  • Nano-Tip A-J: 0.25          Nanoliter
  • Pico-Tip J:       0.06          Nanoliter
  • Nano-Tip J-H: 0.35…0.4 Nanoliter (External heater unit required)
  • Pico-Tip J-H:   0.06          Nanoliter (External heater unit required)

Tips with special design:

  • Nano-Tip HV-J: For high-viscous samples
  • Nano-Tip AR-J: Slim design,for array printing into 96-well plates

 

 

Fast solenoid valve on the Nano-Plotter (SOF mode)

 

Solenoid Valve Pipetting Tips (Option):

  • 40…350 Nanolitre drop volume
  • Affordable glass nozzle for low operational costs
  • Upgrades available for all Nano-Plotter types

 

 

Liquid Management for Piezoelectric Tips

 

The Nano-Plotter piezoelectric pipetting tips consist of glass and silicon.

Nano-Plotter – Removable Slide Tray

Removable Slide Tray

For fast and easy replenishment of targets. A coolable slide tray slows down spot evaporation for higher spot quality.
Different layouts (coolable/non-coolable, with/without locator pins, for slides/MTPs, customized) available for both
NP 1.2 and NP 2.1.

 

nanoplotter-slide-deck-chilled-platform-gesim-1 Nanoplotter - a coolable slide tray slows down spot evaporation for higher spot quality

Nano-Plotter – Microplate Handler

Microplate Handler

Nanoplotter- microplate handler Cytomatn c425

The  Nano-Plotter (NP2.x only) hooks up to the plate hotel Cytomat 2 C425® (ThermoFisher Scientific). The plate hotel is humidity and temperature conditioned and deliveres up to 42 plates on request, thus enabling unattended runs of a large set of samples.

 

 

Nano-Plotter – Chilling and Humidifying

Chilling and Humidifying

These components are to tune the operational environment of slides and source plates. Evaporational losses can weaken the results of microarray experiments.

 

The ultrasonic humidifier blows an aerosol into the dust protection cover of the Nano-Plotter. The sensor controlled add-on unit increases the humidity smoothly up to 80%.

 

The recirculation bath (another add-on unit) connects to the slide deck as well as to the holder of the micro plate.

 

Nanoplotter Chiller nanoplotter-humidifiers-gesim nanoplotter-mtp-holder-gesim

Target Trays

Customized Trays are available on Request

For fast and easy replenishment of targets, standard Nano-Plotter trays adopt both 1" by 3" slides and membranes. Four strips with locator pins separate the slides from each other and inversion of the strips gives a smooth area for membrane arrangement.

Customized shapes are available on request.
SlideTray

Chilling/ Humidifying

Management of Rare Sample Species

External units for chilling and humidifying allow establishment of conditions close to the dew point. The ultrasonic humidifier connects directly to the hood enclosure of the instrument, thus even microliters of samples can remain on the deck of the instrument with minimum evaporation losses.

Coolable_tray_microwell_plate

Both the coolable base plate underneath the slide tray and the microplate holder can be connected to a recirculating chiller to control the temperatures of both the samples and the targets.

 

Benefits

Optical Target Recognition

The Optical Target Recognition (Option) comprises a camera system assembled to the print head as well as a software extension for the Nano-Plotter software.

Considering a typical drop diameter of 50 Microns the Nano-Plotter is well appropriated to apply Nanolitre amounts of liquid to target structures in the same range. However, the nozzle alignment above tiny targets may become challenging if attempted manually.

Microfluidic Chip with tiny dispense targets

Patterned chip layout

Alignment Camera for automatic nozzle adjustment

Alignment camera

The Optical Target Recognition automatically detects small patterns such as microelectrodes, nanowells and microcantilevers. This enables the Nano-Plotter to precisely find positions and spot onto them (manual selection of points by clicking in the camera window is also possible).

The camera looks for two alignment marks (“Fiducials”) on each chip/substrate. Spotting positions can be defined relative to the alignment marks. Regular target patterns like arrays of small wells usually don’t require dedicated fiducials.


Click here for a customer example…!

 

Purpose and Benefit

How does it work?

Microfluidic channel carriers can become expensive and complicated. On the other hand, microfluidic chips are sensitive and just for single use. E.g. diagnostic stuff for routine use typically features hot embossed microplastics, a procedure which initially requires expensive casting machines and layout specific casting tools.

Experimental Setup of a MicCell with Microscope and Accessories

Experimental Setup of a MicCell with Microscope and Accessories

 

 

 

For R/D purposes the MicCell combines an affordable casting process using PDMS and a convenient mechanical setup for quick replacement of the used PDMS channel plate. The mechanical frame of the MicCell fits on standard microscope stages.

 

MicCell – An Easy to Use Microperfusion System

Introduction

Core of the MicCell is an optical transparent PDMS (Polydimethylsiloxane) chip with a user specific microfluidic channel layout. It is accomplished by “channel spacers” for fluidic interconnections and calottes/ polycarbonate frames for mechanical adaptation to standard microscope systems.

The "MicCell-Process": Preparation of the PDMS-CP "Channel Plate" (Left); Position of the PDMS-CP during usage (Right)

The “MicCell-Process”: Preparation of the PDMS-CP “Channel Plate” (Left); Position of the PDMS-CP during usage (Right)

The disposable PDMS chip can be done by the customer using the included casting set. It is easily replacable, therefore microfluidic experiments can be done at affordable costs.

Standard sizes for the PDMS channel plate are:

  • 22 x 22 mm
  • 22 x 50 mm
  • 25 x 75 mm

The PDMS channel plate goes in between the Polycarbonate carrier and the calotte and gets sealed by a standard glass lid. Glass lids wih electrode structures on the inner side are available on request.

On top of all, the MicCell channel plate carrier can be completed with other microfluidic parts, tubes, filters, bottles, syringe systems and an operation software. Move on to Options for more information.


The flyer on the right side provides an overview of the MicCell system. The catalogue reveals technical specs and all accessories along with ordering information.

 

 

Casting Set

How to make the PDMS Channel Plate

The casting set mainly comprises the casting station, Polycarbonate carriers, liquid PDMS and syringes. In addition, you will need a “Master Chip” introducing your particular channel layout.

PDMS-CP Casting Set (Left) with Casting Station (Right) and Accessories

PDMS-CP Casting Set (Left) with Casting Station (Right) and Accessories

The master chip is made from Teflon coated silicon and made at GeSiM on request. Basically we accept any layout provided by a CAD drawing; a set of typical channel layouts is on the shelf. Please ask.

 

Screwable Microvalve

The Hydrogel Valve – Properties and Function

To add liquid, to start and stop reactions, a dead volume free microvalve (GeSiM patent) can be placed into the flow system.

Hydrogel Microvalve HG-7

Hydrogel Microvalve HG-7

HG-7 Microvalve on a MicCell Carrier

HG-7 Microvalve on a MicCell Carrier

The valve is made from a silicon chip but comes with different housings for convenient handling. Most popular version is called “HG-7″, based on a PEEK-housing with 1/16” fitting.

The valve chip incorporates a small but well defined amount of a hydrogel immobilized in a liquid transparent chamber. The surrounding heater controls the temperature of the chamber in a limited range around the swelling temperature of the hydrogel.

 

 

 

Switching Characteristics of the GeSiM Hydrogel Valve

Switching Characteristics of the GeSiM Hydrogel Valve

 

For operation of the valve nothing but 250 mW power is required. The Fluid Processor provides full control of the device.

The sample liquid gets in contact with the enclosed hydrogel. Therefore the valve is mainly made for aqueous solutions but tolerates <15 % methanol, acetone.

 

Flow Sensor

Measurement of Tiny Flows

The GeSiM flow sensors has been designed for flow rates in the range of 0…100 Microliters per minute. It is a thermocalorimetric sensor measuring the heat transfer in a very tiny chamber. The fluid sample is negligibly heated whereas the surroundig temperature distribution is monitored.

As larger the heat transport through the chamber as more precisely the measurement result will be. In other terms, as more time is allowed for a single measurement as better the accuracy.

The sensor can be used along with the MicCell Fluid Processor (See next article) but is available for standalone operation too. The sensor is cascadable, it allows to measure several independent fluid flows simultaneously. Up to four stacked sensors can be managed by a single controller. The controller connects to a PC through serial interface, a dedicated software displays the measurement result.


Please download the flyer (right) for more information.

 

FluidProcessor

Controlling of Complex Setups

MicCell FluidProcessor / Miccell System on a Microscope

MicCell FluidProcessor / MicCell System on a Microscope

 

 

 

The GeSiM Fluidprocessor is a racked unit combining several types of valves and syringe pumps. It is usually built to a particular application.

 

 

 

 

 

 

 

 

 

 

 

In addition, a Windows based software (“MicCell-Software”) is available for controlling

  • Fluidprocessor
  • Hydrogel Valves
  • Piezoelectric Dispensers
  • Flow Sensors

 

A Versatile Microfluidic System

Selected Applications

The MicCell System gained much attention from researchers world wide. Here we present a list of applications already done. Please contact us for further details.

  • Micro-reaction technology, e.g., hybridization or stoppedflow chamber, using fluorescence detection
  • Immobilization of biomolecules (e.g., protein or DNA) in the microchannels before assembly, e.g., by microarraying
  • Generation of concentration gradients perpendicular to the microchannel cross section by a “gradient mixer”
  • Semi-automatic drug screening using adherent cells or tissue slices
  • Viability tests and other cell-based physiological assays
  • Measurement of the interaction of cells with immobilized proteins, DNA, RNA, oligo- or polysaccharides, lipids, and other ligands
  • Cell handling and sorting using optical tweezers
  • Identification of cancer or stem cells using an “optical stretcher” (patent University of Leipzig)
  • Electroporation in the flow
  • Testing of the uniformity of microbeads and other particles, potentially with sorting
  • Manipulation of elongated macromolecules (e.g., DNA or motor proteins) in hydrodynamic flow fields for the bottom-up construction of nanostructures, force measurements, etc.
  • Micro-capillary electrophoresis under the microscope
  • Integration of, e.g., column or filtration material for micro-purifying with or without microscope control
  • Liquid processing independent of a microscope (e.g., assays using electrochemical detection)
    Chemical synthesis on the nanoscale
  • Observation of opaque objects in the MicCell using the pivotable sample carrier

 

µContactPrinter 4.1

Flexible and fully automatic patterning Platform

The µContact-Printer 4.1 (µCP 4.1) features PDMS stamps with customized stamp patterns. It offers surface patterning at the submicrometer range as well as Nano-Imprint Lithography on one instrument. Move to Procedures for more information. It fits to standard laboratory benches and is highly customizable.

 

Please click here for full introduction video.

 

Main specs:

  • Dimension: 623 mm x 521 mm x 367 mm (Length x width x height)
  • Stamp rack for up to five stamps
  • Substrate table: Adjustable; repeating accuracy < 5 µm (XYφ), aided by built-in video microscope
  • Spin coating device for homogeneous distribution of liquids on substrates
  • Windows based control software, preinstalled on computer

 

µCP 4.1 – Tools

At least three independent Z-Axes

Print head of µCP 4.1

Print head of µCP 4.1

µCP 4.1 comes with three up to six individually lowering Z-axes. The basic setup offers:

  • µCP print head for soft membrane stamps up to 20 mm by 20 mm, integrated alignment microscope and UV adapter for optical fibres (5 mm diameter)
  • Pneumatic dispenser for the deposition of high-viscous liquids like photoresist SU8 and NOA 81 (Norland), respectively
  • Syringe dispenser for the application of low-viscous inks for micro-contact printing

 

Struts and Capsules

3D-printing of Cell-loaded Alginate Capsules suspended in Hydrogel

Printable biomaterials can benefit from complex compositions: The release of drugs or cell  growth have to be controlled after printing. A group from the Friedrich-Alexander University in Erlangen added prefabricated capsules enriched with cells to hydrogel before printing.

The GeSiM BioScaffolder was part of this study. It presents a novel method to produce macroporous hydrogel scaffolds in combination with cell-loaded capsule-containing struts by 3D bioplotting.

This approach enables loading of the capsules and strut phases with different cells and/or bioactive substances and hence makes compartmentalization within a scaffold possible.

Light microscopy images of cell-loaded alginate capsules in ALP-loaded alginate struts immediately after fabrication. The free space in the center of the image is a macropore.

Light microscopy images of cell-loaded alginate capsules in ALP-loaded alginate struts immediately after fabrication. The free
space in the center of the image is a macropore.

Fluorescence microscopy image after 10 days of culture and OsteoImage®, DAPI and Vybrant staining capsules loaded with ALP. Green: calcium phosphate. Blue: cell nuclei. Red: cell body. Scale bars: a = 200 µm, b = 500 µm

Fluorescence microscopy image after 10 days of culture and OsteoImage®, DAPI and
Vybrant staining capsules loaded with ALP. Green: calcium phosphate. Blue: cell nuclei. Red: cell body. Scale bars: a = 200 µm, b = 500 µm

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

The goal was to produce scaffolds for possible applications in bone tissue engineering consisting of alginate struts containing alginate capsules enriched with MG-63 osteoblast-like cells and ALP (alkaline phosphatase). Two combinations were compared, namely ALP in the struts and cells in the capsules and vice-versa. Both combinations were cytocompatible for cells and mineralization of scaffolds could be detected in both cases, according to an OsteoImage staining. ALP had no adverse effect on cytocompatibility and enhanced mitochondrial activity.

Different components desirable for bone regeneration, e.g., cells and bioactive proteins, can be incorporated both in the capsules and struts. This enables compartmentalization of components, which facilitates greater flexibility in modification of the scaffold.

Institute of Biomaterials Erlangen

Institute of Biomaterials Erlangen


Rainer Detsch, Bapi Sarker, Tobias Zehnder, Aldo R. Boccaccini and Timothy E.L. Douglas:
Additive manufacturing of cell-loaded alginate enriched with alkaline phosphatase for bone tissue engineering application. De Gruyter, BioNanoMat 2014; 15(3-4): 79–87

 

 

Concept of the BioSyntheSizer

Introduction

Introduction video (720p) on Youtube

Introduction video (720p) on Youtube

The compact GESIM BioSyntheSizer manifests a new instrumentation concept for robots for complex laboratory automation on low-volume scale. It is a new, very compact lab automation platform whose goal is utmost flexibility. In the standard configuration it masters several different chemical syntheses simultaneously, as long as they fit in a liquid handling scheme. This, on the other hand, allows parallelization without having to make a new setup for each synthesis.

 

20 years of experience in the field of microfluidics and lab automation sets the BioSyntheSizer apart from common pipetting robots. Hard- and software follow a strictly modular approach and makes the platform open for application specific configurations. We will be pleased to learn about your requirements in chemical synthesis!

 

Multi-Function Head for Free Choice of Tools

Interchangeable Tools for up to 7 Z-Axes

Individually lowering Z-axes allow to synchronize multiple tools with different dimensions. The head of the instrument connects to the F-box (Media control unit for compressed gas, power, vacuum).

 

Example of a BSyS print head with seven Z-axes: (1) camera, (2+3) twin tool vaccum gripper + 3-lumen pipet, (4) FlipTube cap opener, (5) heatable piezo pipet +Z-Sensor on one axis, (6+7) nozzle/needle adapter (From left)

Example of a BSyS print head with seven Z-axes: (1) camera, (2+3) twin tool vaccum gripper + 3-lumen pipet, (4) FlipTube cap opener, (5) heatable piezo pipet +Z-Sensor on one axis, (6+7) nozzle/needle adapter (From left)

 

Liquid Handling

  • Conical receptacle to load/ eject disposable pipet tips and Luer needles
  • Needle tips for penetration of antiseptic lids
  • Piezoelectric Picolitre pipettes
  • Twin-tip Piezoelectric pipet: Two piezo pipets can be swivelled to mix up drops in flight
  • Solenoid dispense valves for the Nanolitre range
  • Pneumatic/ piezo dispense valves for more viscous drops

Mechanical tools

  • Vacuum gripper for crimp vials and 3-lumen pipette for inert gas, solvent, vacuum, e.g. for azeotropic drying of solutes
  • Tool for cap opening/closing of FlipTube microcentrifuge tubes
  • Z-sensor to measure height profiles of substrates

Others

  • Powder pipettes for the Microgram range
  • Cartridge dispensers for high viscous media
  • Titration pipette for automatic adjustment of pH
  • Camera for pattern recognition and QR code identification
  • UV lamp for crosslinking

 

 

Racks and Reactors

Components for the Work Area of the Instrument

BSys3.1 with application specific kit plates onboard (right)

BSys3.1 with application specific kit plates onboard (right)

 

On the work deck you find easily mountable and removable holders for disposable pipette tips, needles, microtitre plates, glass slides, storage and reaction vessels (e.g. crimp vials with septum, microcentrifuge tubes with cap) and other freely definable objects that can be heated or cooled on request. A time-controlled connection to extraction or cleaning columns, an HPLC or a photometric analysis can be done via tube adapters.

 

  • Trays for micro well plates/ slides
  • Racks for microcentrifuge tubes, septum vials, disposable pipette tips, Luer-lock needles
  • Racks  with tube adapters to connect nozzles to tubes/columns etc.
  • Racks with filter cartridges for ion separation
  • Application specific microfluidic manifolds
  • Reactors with antiseptic lid, temperature up to 150°C, pressure up to 8 bar
  • Magnetic stirrers
  • Wash/Dry stations, optical tools for tips alignment and droplet measurement (stroboscope)