Question 1

Dr. Dani Cai  from the National Institutes of Health

Celine Billerit · Accepted Answer

“The BioPen System is one of the best single-cell treatment system on the market. It is easy-to-use, and gives me beautiful, publication-ready movies.”

Question 2

U.S. Food and Drug Administration

Celine Billerit · Accepted Answer

“We have a need for the BioPen System to be used for studying variations in cell contractility due to the exposure of certain drugs or combinations of drugs. [The BioPen] is critical for this effort in acquiring large amounts of data.

It has two advantages:

allows us to save money on expensive drugs because it uses very small amounts;
allows us to save money and resources in stem cell differentiation because the same culture of cells can be used for many measurements.”

Question 3

Prof. Rizzo from the University of Maryland Medical Center

Celine Billerit · Accepted Answer

“The main uses of the Biopen is for stimulating migrating cells. The benefits when using the system is that it can stimulate a portion of a cell. The system provide us with unique data with it you are able to see how cell process are trapped.”

Question 4

Dr. Joseph Bruton from Karolinska Institutet

Celine Billerit · Accepted Answer

“I chose the BioPen System because I can target precise areas of the cell and see how local and global calcium changes in response to different perturbations. The small size of the area means that multiple sites can be examined in a single muscle cell.

The BioPen System has several advantages:

small footprint on the lab bench,
small target area, although this can be enlarged quite a bit,
consumption of expensive antibodies and drugs is minimized compared to perfusion of the whole cell as we used to have to do.”

Question 5

Associate Prof. Kent Jadermark from Karolinska Institutet

Celine Billerit · Accepted Answer

“We have tested the BioPen in different applications here at Karolinska Institutet and in collaboration with a research group at Göteborg University. I consider the BioPen very valuable for applying compounds or drugs to single cells in culture as well as to specific cell layers in brain slices”.

Question 6

Veronica Jove from the Rockefeller University

Celine Billerit · Accepted Answer

«The BioPen product is well designed and works even better than we could have hoped. The scientist who designed the BioPen flew from Sweden to our university to install the product and tailor it to our specific research goals. We have had no installation, device or software problems and have found the product to be flexible for different research purposes. The team is very reliable with customer support. I would highly recommend purchasing this product»

Question 7

Roland Malli's Lab from Medical University of Graz

Celine Billerit · Accepted Answer

"For the application of expensive substances or small amounts of reagents, it’s challenging to use perfusion systems. Using the BioPen System allows us now to precisely perfuse only few single cells and record their subsequent reactions and metabolic changes. In terms of cell-to-cell communication, it has always been impossible to expose a single cell to a stimulus and monitor the reaction of surrounding cells. With the Biopen, it might lead to new, and yet unknown, insights into intercellular communications within cellular networks."

Question 8

Are there any requirements regarding instrumentation, i.e. does this system work with any micromanipulator?

Celine Billerit · Accepted Answer

The BioPen® System integrates with any standard type of micromanipulator capable of holding a 7mm rod. A micromanipulator is required to position the BioPen within the sample dish.

The BioPen system includes:

A box of 10 disposable BioPen pipette tips.
- Two types of tips are available: PRIME and FLEX.
The BioPen holder manifold.
- Two types of BioPen holders are available: PRIME and FLEX.
The PPC1 multichannel pressure controller.
A configured touch-screen tablet computer.
The BioPen Wizard software.

Question 9

Can the BioPen system be used in an upright microscope?

Celine Billerit · Accepted Answer

The BioPen FLEX system has been specially designed to work with upright microscopes, requiring a long working distance objective with at least 11 mm of clearance. Depending on required BioPen arrangement angle, we recommend either using larger sample dish is recommended or a shallow wall height dishes. (Common example being WillCo Wells GWST-5040).

Question 10

Can the BioPen system be used with any objective?

Celine Billerit · Accepted Answer

Yes, in general. On all microscopes, we recommend a lower magnification for initial positioning, which can then be switched to a higher magnification if required. On inverted microscopes, any objective can be used: air or immersion medium, short or long working distance. For upright microscopes long working distance air objectives are required.

Question 11

Can the BioPen system be used with other probes?

Celine Billerit · Accepted Answer

Yes, the BioPen System has been optimized to fit into practically any micromanipulation environment, allowing combination with other probes such as patch-clamp pipettes, optical fibers or electrodes. The BioPen FLEX system, expands the available space around the sample allowing for easier positioning of multiple probes

Question 12

Is it possible to integrate with other equipment or have external triggers?

Celine Billerit · Accepted Answer

The PPC1 is equipped with both input and output communication capability, through the use of standard 5V TTL signals. Protocols can therefore be initiated by external equipment (such as a camera or amplifier) or trigger signal can be sent to external equipment. The “Rise” or the “Fall” of the trigger can be used, as well as the duration and wait periods can be set within the BioPen Wizard.

Question 13

Can the BioPen system be used within a microscope incubator chamber?

Celine Billerit · Accepted Answer

Yes, the BioPen system can be used within a microscope incubator chamber. The BioPen holder is made from high-quality stainless-steel components and plastics capable of withstanding a multitude of environments.

Question 14

Can the BioPen system be used in sensitive electronics recording environments?

Celine Billerit · Accepted Answer

The BioPen system can be used in the vicinity and in conjunction to a sensitive electronic detection and recording devices because it is driven by the air pressure, therefore no conductive medium connects the BioPen holder to the surrounding environment.

Question 15

How long does it take to setup the system?

Celine Billerit · Accepted Answer

To setup the BioPen system and to position correctly the BioPen pipette tip into your sample, it requires approximately 15 minutes. Once the micromanipulator is pre-positioned, future setup times can be shorter. Using established protocols, the priming time for the pipette tip is approximately 5 minutes.

Question 16

How much and how many solutions can I put in one BioPen pipette tip?

Celine Billerit · Accepted Answer

Each BioPen pipette tip can be loaded with up to 4 different solutions at one time. The BioPen pipette tip contains 8 wells: 4 wells containing solutions for delivery and 4 wells for collection of waste.

Each well can contain up to 35 μl of solution, although 20-30 μl is recommended for ease of handling. Less than 10 μl of solution is not recommended, as it is difficult to purge the BioPen pipette tip with such little solution in the well. Liquid must be placed into all 4 solution wells. If fewer than 4 solutions are needed, the remaining solution wells should be filled with a buffer or alternative aqueous solution.

Question 17

What types of solutions are compatible with the BioPen pipette tips? Does it work with high viscosity solutions?

Celine Billerit · Accepted Answer

BioPen pipette tips are designed to work with any aqueous solution, as long as all 4 solution wells are loaded. All of the loaded solutions should be of approximately the same viscosity to minimize variances in the delivery region when switching between the loaded solutions.

BioPen pipette tip can be used with solutions of a viscosity between 1- 500 cP, however higher viscosity solutions should be carefully degassed prior to use, to minimize the possibility of trapping air bubbles within a BioPen tip.

Question 18

Does the BioPen pipette tip have any dead volume?

Celine Billerit · Accepted Answer

The BioPen pipette tip is designed to have the solution meeting point very close to the sample, almost eliminating any dead volume. The remaining small volume is approximately 0.4 nL for the BioPen pipette tip PRIME and 1.5 nL for the BioPen pipette tip FLEX.

Question 19

What are the BioPen pipette tips made of?

Celine Billerit · Accepted Answer

The BioPen pipette tips are made of polydimethylsiloxane (PDMS), a flexible medically approved elastomer material. A thin polypropylene film protects the back of the tips when loading into the holder.

The BioPen pipette tips, unlike glass pipettes, won’t break upon contact with a hard surface, and accidental contact will not impair the function.

Question 20

What are the advantages of using a disposable fluidic pipette tip?

Celine Billerit · Accepted Answer

Very minimal clean up times, no cross contamination between experiments (both of solutions and sample dish contaminants) are key advantages to the disposable pipette tips. Sterile pipette tips are also available as an option upon request.

Question 21

How often do the BioPen pipette tips need replacing?

Celine Billerit · Accepted Answer

To offer a contamination free controlled delivery system, the BioPen pipette tips are designed to be one-time use. Typical usage is up to 6-8 hours, with occasional emptying of the waste wells.

Question 22

Are BioPen pipette tips sterile?

Celine Billerit · Accepted Answer

Standard BioPen pipette tips are produced in a high precision, low particle environment, using clean room protocols. They undergo multiple stages of visual and solution injection-based quality control steps. The standard BioPen pipette tips are not sterilized, but sterile pipette tips can be offered as an add-on service option, using an independently validated process (RISE).

Question 23

How many BioPen pipette tips are in a box?

Celine Billerit · Accepted Answer

Each box contains 10 BioPen pipette tips of either the PRIME or FLEX kind.

Question 24

The BioPen is a hydrodynamic confined flow device. What is an hydrodynamic confined flow and what does it look like?

Celine Billerit · Accepted Answer

A hydrodynamic confined flow device is essentially a flow chamber, where a small volume of fluid is spatially confined within the other, significantly larger fluid volume.

The two miscible or immiscible liquids are physically in contact yet separated by means of a virtual boundary. Mixing between the two zones is largely defined by diffusion. The experimental conditions necessary to achieve efficient confinement, while inhibiting diffusion across the virtual boundary, are characterized by low Reynolds and high Péclet numbers. The hydrodynamic flow is a continuous steady state flow with minor fluctuations due to the pressure pump feedback control loop that generates a stable and continuous pressure.

Question 25

How does the delivery of a compound remain localized and does not diffuse to other areas?

Celine Billerit · Accepted Answer

The BioPen tip has 3 channels at its front, introduced into the open volume:

The central channel serves as an injection port or output channel (positive pressure), where liquid is delivered to the open volume.
Two adjacent channels function as aspiration ports or input channels (negative pressure), through which liquid is drawn in from the sample dish (see figure below).

Perfusion recirculation — *Schematic of the hydrodynamic flow formed at the end of the pipette tip*

Balancing this inflow and outflow, creates a small perfusion volume or delivery zone at the front of the BioPen tip.

To generate a localized delivery zone, the flow rate of the aspiration port has to be higher than the flow rate of the injection port, such that all fluid leaving the outlet would flow entirely into the neighboring inlet. Flow recirculation provides a mean to maintain a localized delivery volume, which would otherwise rapidly fade by diffusion and contaminate the surroundings.

Standard flow conditions are set to have 10 times more inflow than outflow.

Question 26

How big is the area/volume of the delivery zone?

Celine Billerit · Accepted Answer

The size range of the delivery zone is:

20-160 mm long
90-110 mm wide

In volumes this relates to approximately 0.1 to 1 nL.

Question 27

Are the concentration of the solutions in the wells and the concentration of the solutions delivered the same?

Celine Billerit · Accepted Answer

The BioPen is flowing compounds directly from the solution well, through an intricate set of fluidic channels, directly to the sample. There is no dilution within the BioPen, therefore the concentrations are the same.

Question 28

How do we visualize the delivery zone?

Celine Billerit · Accepted Answer

You can visualize the delivery zone using a flow tracer. We recommend Erythrosin B as it is a non-toxic member of the family of fluorescein dyes. It can be easily dissolved in almost any aqueous buffer and is both a colorimetric and fluorescent dye, therefore applicable for brightfield and fluorescent microscopes.

Question 29

Can the delivery be stopped and started again?

Celine Billerit · Accepted Answer

Yes, the delivery can be stopped and restarted again at any time.

Question 30

What is the fastest time the solution can be exchanged within the delivery zone?

Celine Billerit · Accepted Answer

This depends on the BioPen setup; solution speed, positioning to the sample, etc. Under typical operation, complete exchange time within the zone, is less than 1 second.

Question 31

Can any type of cell be employed? What type of cells can be used?

Celine Billerit · Accepted Answer

Any type of cells can be used as long as they are surface adherent cells or held by a probe e.g. patch-clamped cells.

Question 32

Can several cells be targeted at once?

Celine Billerit · Accepted Answer

Yes, you can target one or a small number of cells. The number of cells that you can target depends of the size of your target area. For complete exposure, the target area should not exceed the delivery zone size, that is created by the BioPen system.

You can adjust the size of the delivery zone via the BioPen software. The size of the delivery zone varies between:

20-160 μm in length
90-110 μm in width

Question 33

How is the cell affected by the flow?

Celine Billerit · Accepted Answer

The cell is affected minimally by the flow since the BioPen System creates negligible flow shear stress.

Question 34

What are the main application areas for the BioPen system?

Celine Billerit · Accepted Answer

The BioPen is a very flexible tool for localized compound delivery. It can be employed almost anywhere when there is a need to deliver compounds to confined area and volume, especially useful when delivering compounds to individual cells.

The BioPen has several applications such as:

single-cell enzymology
ion channel kinetics
cell to cell molecular transport
single muscle fiber physiology etc…

Download the Application Notes for more details.

Question 35

Do you have any publications using the BioPen system?

Celine Billerit · Accepted Answer

Yes, there have been numerous scientific publications using the BioPen system, you can find the list on the publications tab,

Question 36

What are the key features of the software?

Celine Billerit · Accepted Answer

The BioPen Wizard is the control software for the BioPen system. It enables users of all experience levels to have direct control over spatial and temporal test solution delivery to individual or group of cells.

Software includes several additional features such as:

Experiment presets and user configuration files
Immediate access to frequently used settings
Interactive tooltips to help users navigate, with instant contextual support
Multiple interface language choices
A scripting module for automation of common task
A simulation mode to design and test protocols before use
A calculation panel to give real-time updates on flow rates within the BioPen tip

Question 37

Does it work on any computer? What are system requirements?

Celine Billerit · Accepted Answer

It works with any PC running Microsoft Windows 10 and 7 (both 32 or 64 bit) with an available USB 2.0 (or above) port.

Authors	Title	Journal	Year	doi
Chinmay Katke, Esteban Pedrueza-Villalmanzo, Karolina Spustova, Ruslan Ryskulov, C. Nadir Kaplan, and Irep Gözen	Colony-like Protocell Superstructures	ACS Nano	2023	10.1021/acsnano.2c08093
Sebastian Persson, Niek Welkenhuysen, Sviatlana Shashkova, Samuel Wiqvist, Patrick Reith, Gregor W. Schmidt, Umberto Picchini, Marija Cvijovic	Scalable and flexible inference framework for stochastic dynamic single-cell models	PLOS Computational Biology	2022	10.1371/journal.pcbi.1010082
Elif Köksal, Inga Põldsalu, Henrik Friis, Stephen Mojzsis, Martin Bizzarro, Irep Gözen	Spontaneous formation of prebiotic compartment colonies on Hadean Earth and pre-Noachian Mars	ChemSystemsChem	2021	10.1002/syst.202100040
Inga Põldsalu, Elif Senem Köksal and Irep Gözen	Mixed fatty acid-phospholipid protocell networks	Phys. Chem. Chem. Phys.	2021	10.1039/D1CP03832J
Sandra Burgstaller, Helmut Bischof, Thomas Rauter, Tony Schmidt, Rainer Schindl, Silke Patz, Bernhard Groschup, Severin Filser, Lucas van den Boom, Philipp Sasse, Robert Lukowski, Nikolaus Plesnila, Wolfgang F. Graier, and Roland Malli	Immobilization of Recombinant Fluorescent Biosensors Permits Imaging of Extracellular Ion Signals	ACS Sensors	2021	10.1021/acssensors.1c01369
Xiaohuan Wang, Long Li, Yingfeng Shao, Jiachen Wei, Ruopu Song, Songjie Zheng, Yuqiao Li, and Fan Song	Effects of the Laplace pressure on the cells during cytokinesis	iScience	2021	10.1016/j.isci.2021.102945
Trkulja, Carolina L., Jungholm, Oscar, Davidson, Max, Jardemark, Kent, Marcus, Monica M., Hägglund, Jessica,Karlsson, Anders, Karlsson, Roger, Bruton, Joseph, Ivarsson, Niklas,Srinivasa, Sreesha P., Cavallin, Alexandra, Svensson, Peder, Jeffries, Gavin D. M., Christakopoulou, Maria-Nefeli, Reymer, Anna, Ashok, Anaswara, Willman, Gabriella, Papadia, Daniela, Johnsson, Emma, Orwar, Owe	Rational antibody design for undruggable targets using kinetically controlled biomolecular probes	Science Advances	2021	10.1126/sciadv.abe6397
Karolina Spustova, Elif Senem Köksalm Alar Ainlam Irep Gözen	Subcompartmentalization and Pseudo‐Division of Model Protocells	Small	2021	10.1002/smll.202005320
Jové, V., Zhongyan, G., Hol, F.J.H., Zhao, Z., Sorrells, T.R., Carroll, T.S., Prakash, M., McBride, C.S., Vosshall, L.B.	Sensory Discrimination of Blood and Floral Nectar by Aedes aegypti Mosquitoes	Neuron	2020	10.1016/j.neuron.2020.09.019
Köksal, E.S., Liese, S., Xue, L., Ryskulov, R., Viitala, L., Carlson, A., Gözen, I.	Rapid Growth and Fusion of Protocells in Surface‐Adhered Membrane Networks	Small	2020	10.1002/smll.202002529
Perpiñá-Viciano, C., Işbilir, A., Zarca, A., Caspar, B., Kilpatrick, LE., Hill, SJ., Smit, MJ., Lohse, MJ., and Hoffmann, C.	Kinetic Analysis of the Early Signaling Steps of the Human Chemokine Receptor CXCR4	Molecular Pharmacology	2020	10.1124/mol.119.118448
Jõemetsa, S., Spustova, K., Kustanovich, K., Ainla, A., Schindler, S., Eigler, S., . . . Gözen, I.	Molecular Lipid Films on Microengineering Materials.	Langmuir	2019	10.1021/acs.langmuir.9b01120
Köksal, E., Liese, S., Kantarci, I., Olsson, R., Carlson, A., & Gözen, I.	Nanotube-Mediated Path to Protocell Formation.	ACS Nano	2019	10.1021/acsnano.9b01646
Orwick Rydmark, M., Christensen, M., Köksal, E., Kantarci, I., Kustanovich, K., Yantchev, V., . . . Gözen, I.	Styrene maleic acid copolymer induces pores in biomembranes.	Soft Matter	2019	10.1039/c9sm01407a
Markwardt, M., Snell, N., Guo, M., Wu, Y., Christensen, R., Liu, H., . . . Rizzo, M.	A Genetically Encoded Biosensor Strategy for Quantifying Non-muscle Myosin II Phosphorylation Dynamics in Living Cells and Organisms.	Cell Reports	2018	10.1016/j.celrep.2018.06.088.
Wright, S., Consuelo, M., Cañizal, A., Benkel, T., Simon, K., Le Gouill, C., . . . Hoffmann, C	FZD5 is a Gαq-coupled receptor that exhibits the functional hallmarks of prototypical GPCRs	Science Signaling	2018	10.1126/scisignal.aar5536
Oehler, B., Mohammadi, M., Viciano, C., Hackel, D., Hoffmann, C., Brack, A., & Rittner, H.	Peripheral interaction of resolvin D1 and E1 with opioid receptor antagonists for antinociception in inflammatory pain in rats.	Frontiers in Molecular Neuroscience	2017	10.3389/fnmol.2017.00242
Waxse, B., Sengupta, P., Hesketh, G., Lippincott-Schwartz, J., & Buss, F.	Myosin VI facilitates connexin 43 gap junction accretion.	Journal of Cell Science	2017	10.1242/jcs.199083
Gözen, I., Jeffries, G., Lobovkina, T., Celauro, E., Shaali, M., Doosti, B., & Jesorka, A.	Single-Cell Analysis with the BioPen	Open Space Microfluidics (Book)	2016	10.1002/9783527696789.ch10
Ahemaiti, A., Wigström, H., Ainla, A., Jeffries, G., Orwar, O., Jesorka, A., & Jardemark, K.	Spatial characterization of a multifunctional pipette for drug delivery in hippocampal brain slices.	Journal of Neuroscience Methods	2015	10.1016/j.jneumeth.2014.12.017
Xu, S., Kim, A., Jeffries, G., & Jesorka, A.	A rapid microfluidic technique for integrated viability determination of adherent single cells.	Analytical and Bioanalytical Chemistry	2015	10.1007/s00216-014-8364-9
Zhang, H., Kim, A., Xu, S., Jeffries, G., & Jesorka, A.	Cellular communication via directed protrusion growth: Critical length-scales and membrane morphology.	Nano Communication Networks	2015	10.1016/j.nancom.2015.10.001
Zhang, H., Xu, S., Jeffries, G., Orwar, O.	Artificial nanotube connections and transport of molecular cargo between mammalian cells.	Nano Communication Networks	2013	10.1016/j.nancom.2013.08.006
Bruton, J., Jeffries, G., & Westerblad, H.	Usage of a localised microflow device to show that mitochondrial networks are not extensive in skeletal muscle fibres.	PLoS ONE	2014	10.1371/journal.pone.0108601
Ahemaiti, A., Ainla, A., Jeffries, G., Wigström, H., Orwar, O., Jesorka, A., & Jardemark, K.	A multifunctional pipette for localized drug administration to brain slices.	Journal of Neuroscience Methods	2013	10.1016/j.jneumeth.2013.08.012
Ainla, A., Gözen, I., Hakonen, B., & Jesorka, A.	Lab on a Biomembrane: Rapid prototyping and manipulation of 2D fluidic lipid bilayers circuits.	Scientific Reports	2013	10.1038/srep02743
Jansson, E., Trkulja, C., Ahemaiti, A., Millingen, M., Dm Jeffries, G., Jardemark, K., & Orwar, O.	Effect of cholesterol depletion on the pore dilation of TRPV1.	Molecular Pain	2013	10.1186%2F1744-8069-9-1
Wegrzyn, I., Ainla, A., Jeffries, G., & Jesorka, A	An optofluidic temperature probe.	Sensors	2013	10.3390/s130404289
Ainla, A., Jeffries, G., Brune, R., Orwar, O., & Jesorka, A.	A multifunctional pipette.	Lab on a Chip	2012	10.1039/c2lc20906c
Ainla, A., Xu, S., Sanchez, N., Jeffries, G., & Jesorka, A.	Single-cell electroporation using a multifunctional pipette.	Lab on a Chip	2012	10.1039/c2lc40563f
Ainla, A., Jansson, E., Stepanyants, N., Orwar, O., & Jesorka, A.	A microfluidic pipette for single-cell pharmacology.	Analytical Chemistry	2010	10.1021/ac100480f

BioPen® SYSTEMS

MICROFLUIDIC SYSTEMS FOR TARGETING SINGLE CELLS

Control the microenvironment of your cells by targeting one cell at the time

COMPARE BIOPEN PRIME AND BIOPEN FLEX

List of publications using BioPen

Dr. Dani Cai from the National Institutes of Health