Pico-Surf Novel surfactants for generation of robust picodroplets
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Pico-Surf™
Novel surfactants for generation of
robust picodroplets
product datasheet page
Description 3
Main Benefits 4
Choosing which formulation to use 4
Using Pico-Surf™ 6
PCR using Pico-Surf™ 1 7
ESI-MS using Pico-Surf™ 2 10
Sphere Fluidics Ltd. 11
The Dolomite Centre Limited Page 1 of 12
The Dolomite Centre Ltd Part name Part No. Pico-Surf™ 1, 10ml, 2% in FC-40 3200202 Pico-Surf™ 1, 10ml, 5% in FC-40 3200203 Pico-Surf™ 1, 50ml, 2% in FC-40 3200204 Pico-Surf™ 1, 50ml, 5% in FC-40 3200205 Pico-Surf™ 1, 10ml, 2% in Novec-7500 3200211 Pico-Surf™ 1, 10ml, 5% in Novec-7500 3200214 Pico-Surf™ 1, 50ml, 2% in Novec-7500 3200215 Pico-Surf™ 1, 50ml, 5% in Novec-7500 3200216 Pico-Surf™ 2, 10ml, 2% in FC-40 3200219 Pico-Surf™ 2, 10ml, 5% in FC-40 3200220 Pico-Surf™ 2, 50ml, 2% in FC-40 3200221 Pico-Surf™ 2, 50ml, 5% in FC-40 3200222 Pico-Surf™ 2, 10ml, 5% in Novec-7500 3200278 Pico-Surf™ 2, 50ml, 5% in Novec-7500 3200280 Pico-Surf™ 2, 10ml, 2% in Novec-7500 3200281 Pico-Surf™ 2, 50ml, 2% in Novec-7500 3200282 MAR-000124_v.A.14 Page 2 of 12
The Dolomite Centre Ltd
Description
Picodroplets are a fascinating area of research. They are especially important where tests
need to be conducted on only a few nanoliters of sample containing, for example, cells or
DNA. Picodroplet technology enables scientists and researchers to perform thousands of
simultaneous reactions on single cells and small populations of molecules contained
within aqueous droplets.
Pico-Surf™ 1 & 2 are specialised chemical solutions
containing a proprietary surfactant in a fluorocarbon
carrier oil. The surfactant acts as an interface
stabilization agent that enables stability of the
picodroplets under a wide range of temperature and
biological conditions, as well as allowing picodroplets to
be stored for several weeks in suitable conditions.
The Pico-Surf™ surfactants were developed by Sphere
Fluidics and are sold exclusively by Dolomite, as part of
a technical and commercial collaboration between the
two leading companies. Pico-Surf™ 1 is specially
designed for generating aqueous solution-in-oil
picodroplets for molecular biology assays, for instance,
polymerase chain reaction (PCR) and other isothermal
DNA amplification reactions. Pico-Surf™ 2 is more
suitable for applications involving cell growth or
electrospray ionisation mass spectrometry (ESI-MS).
For other applications, please contact Dolomite directly
for advice on which surfactant would be the most
suitable option to use.
Both versions are available pre-dissolved at 2% and
5% concentration in either of two fluorocarbon oils from
3M Company which are commonly used in microfluidic
droplet generation:
FC-40™ Fluorinert™
Novec™ 7500
MAR-000124_v.A.14 Page 3 of 12
The Dolomite Centre Ltd
Please note: For PCR thermocycling applications, we only recommend using Pico-
Surf™ 1 with Novec™ 7500. FC-40 is not recommended for this application.
Available in 10ml and 50ml, the surfactant can be used together with Dolomite’s Droplet
Junction Chips as well as Dolomite’s novel Picodroplet Formation Chip and Picodroplet
Reservoir Chip. It can also be used with the Pico-GenTM Picodroplet Formation Biochips
from Sphere Fluidics. For best results we recommend using with a flurophilic surface
coating.
Main Benefits
Pico-Surf™ 1
Can generate aqueous solution-in-oil droplets for molecular biological assays, for
instance, polymerase chain reaction (PCR) and other isothermal DNA
amplification reactions.
Facilitates reliable formation and stabilization of droplets from Ø 25µm to Ø 100µm
(volume range 8pl to 500pl)
Long shelf life
High purity
Can be used directly, no further formulation required
Pico-Surf™ 2
Suitable for cell growth studies and analysis by electrospray ionisation mass
spectrometry (ESI-MS)
Facilitates reliable formation and stabilization of droplets from Ø 25µm to Ø 100µm
(volume range 8pl to 500pl)
No or low background signal in electrospray mass spectrometry (ESI-MS)
Long shelf life
High purity
Choosing which formulation to use
Pico-Surf™ 1 Pico-Surf™ 2
Recommended for molecular biology Recommended for cell growth studies
assays such as digital polymerase chain and electrospray ionisation mass
reaction (digital PCR). spectrometry (ESI-MS).
MAR-000124_v.A.14 Page 4 of 12The Dolomite Centre Ltd
Storage
Pico-Surf™ 1 & 2 should be stored in the dark at room temperature. Do not refrigerate.
Carrier fluid
Pico-Surf™ 1 & 2 are both available in Novec-7500 and in FC-40. Deciding which carrier
fluid to choose will be variable depending on the specific application and some
experimentation may be required.
Please note: For PCR thermocycling applications, we only recommend using Pico-
Surf™ 1 with Novec™ 7500. FC-40 is not recommended for this application.
Novec-7500 has a much lower resistivity than FC-40. Resistivity of Novec-7500 is 2.2 x
10^8 ohm-cm and resistivity of FC-40 is 4.0 x10^15 ohm-cm. Novec-7500 also has a
lower viscosity (0.8CSt versus 2.2CSt) which would work well with flow resistors to
balance with water continuous phase. Below is a summary list of physical properties of
Novec-7500 and FC-40.
Novec-7500 FC-40
Boiling point (°C) 128 155
Pour Point (°C) -100 -57
Molecular Weight (g/mol) 414 650
Critical Temperature (°C) 261 270
Critical Pressure (Mpa) 1.55 1.18
Vapour Pressure (kPa) 2.1 0.43
Heat of Vaporization (kJ/kg) 89 68
Liquid Density (kg/m3) 1614 1850
Coefficient of Expansion (K-1) 0.0013 0.0012
Kinematic Viscosity (cSt) 0.77 1.8
Absolute Viscosity (cP) 1.24 3.4
Specific Heat (J/kg-K) 1128 1100
Thermal Conductivity (W/m-K) 0.065 0.065
Surface Tension (mN/m) 16.2 16
Solubility of Water in Fluid (ppm
by weight) 45The Dolomite Centre Ltd Concentration Pico-Surf™ 1 & 2 are both available in 2% and 5% concentrations. Surfactant concentration requirements are variable dependent on the specific application and some experimentation may be required. Concentration can be modified by diluting with the appropriate carrier fluid. Filtration: Pico-Surf is filtered in solution after production using a 0.1 µm polyethersulfone (PES) filter. Each sample is stored in a bottle which has been cleaned with nitrogen gas to remove dust particulates. When the product is first used, additional filtration is not required. Filtration may be necessary after the product has been opened, as exposure to dust particulates may contaminate the sample Using Pico-Surf™ Droplet Storage and Incubation Users are recommended to store droplets generated with Pico-Surf™ in sterile, plastic (micro) centrifuge tubes. However, they can also be stored in any device or container having hydrophobic surface properties. Once generated, Pico-Surf™ emulsions are stable for up to one week when stored between 4°C to 65°C. For long-term storage, especially at elevated temperatures, topping the emulsion with mineral oil is recommended to reduce or eliminate evaporation and contamination. Droplet size The performance of the surfactant depends, amongst other things, on the size of the droplet and properties of the aqueous phase. As droplet size increases, coalescence becomes more likely, especially under, e.g. PCR thermal cycle conditions. As a guide, 100µm (700pl) is an absolute maximum for room temperature use, 30µm in diameter (15pL) or smaller are recommended for PCR. The user may need to test for optimal droplet size based on specific application and reagents used. MAR-000124_v.A.14 Page 6 of 12
The Dolomite Centre Ltd
Product Filtration
Filtration may be necessary after the product has been opened, as exposure to dust
particulates may contaminate the sample. Polyethersulfone (PES) membranes with a
membrane size of 0.2 µm or 0.1 µm are recommended. Cellulose acetate membranes of
the same size are also suitable. PTFE membranes are not recommended as a high back
pressure is created which can affect the performance of the product.
PCR using Pico-Surf™ 1
Below is a typical example of PCR reactions in picodroplets generated from Pico-Surf-1.
Materials and Reagents
Pico-Surf™ 1, 5% in HFE-7500 (Sphere Fluidics Ltd).
Commercial PCR kit 1.
Commercial PCR kit 2.
Template: Human genomic DNA
Primers: ACTB primer set.
Key experimental conditions
Picodroplet size: 59 micrometres
DNA concentration: 16 ng/uL
Primer concentration: 0.3 uM
Thermal cycle conditions: 95°C, 2 min, hot start
30× 95°C, 10s
59°C, 30s
72°C, 30s
10°C, hold
3. Typical experimental results:
a) Thermal stability:
The picodroplets were inspected under microscope to verify their integrity after PCR
thermal cycle (see image below). Although larger picodroplets might be applicable
under certain experimental conditions and application requirement, it is recommended
to generate smaller than 30 micron droplets for emulsion PCR, to ensure integrity of
picodroplets after PCR thermal cycles.
MAR-000124_v.A.14 Page 7 of 12The Dolomite Centre Ltd b) PCR results: PCR product can be recovered by breaking emulsion using Pico-Break™ reagents then analysed by gel electrophoresis. PCR in a conventional format was also tested as control. Gel electrophoresis result below. Note: Pico-droplet integrity or PCR efficiency may vary when using different commercial PCR reaction kits. The user should optimize the choice of reagents based on their needs. MAR-000124_v.A.14 Page 8 of 12
The Dolomite Centre Ltd
Result of gel electrophoresis
General guidance for PCR
Pico-Surf™ 1 emulsion is stable under most PCR thermal cycling conditions. The
following tips should be considered when optimising your specific PCR reaction.
Generate picodroplets smaller than 15pL in volume (30µm in diameter).
Top the emulsion with mineral oil if possible.
Mainly to prevent protein aggregation, detergent addition is sometimes included in
commercially-available PCR reagent mixtures or recommended in published
papers or laboratory protocols. Depending on their composition and concentration,
these detergents can substantially deteriorate droplet stability under the conditions
used in thermal cycling. Users should titrate the detergent in their PCR solution
and also optimise this with reference to the Pico-Surf™ 1 concentration and other
assay parameters.
MAR-000124_v.A.14 Page 9 of 12The Dolomite Centre Ltd
ESI-MS using Pico-Surf™ 2
The following graphs show a comparison of protein analysis in samples analysed using
picodroplets ESI-MS in contrast with conventional ESI-MS.
alpha LA droplets
XL 20110809 alpha lactalbumin 20 uM droplet 9 53 (0.134) Cm (3:561) TOF MS ES+
2024 290
100
2023 2024 Picodroplet MS:
2024
2023 2027 20 M -lactalbumin in 22.5 mM NH4OAc
%
2018 2028 2% Pico-Surf 2 in HFE3283
2018
0
XL 20110809 alpha lactalbumin 20 uM conventional 8 (0.148) Cm (6:13) TOF MS ES+
2024 313
100
2027 Conventional MS:
2023 2027 20 M -lactalbumin in 22.5 mM NH4OAc
%
2023 2030
2021
0 m/z
1000 2000 3000 4000 5000 6000 7000
Pico-Surf™ 2 is also well suited to cell growth studies whereby single cells are
encapsulated in droplets for examination of growth behaviour.
Microalgae growth in picodroplets stabilized with Pico-Surf™ 2
MAR-000124_v.A.14 Page 10 of 12The Dolomite Centre Ltd Sphere Fluidics Ltd Sphere Fluidics Ltd is a leading company specialised in picodroplet technology that performs thousands of simultaneous analyses on single cells and small populations of molecules. Sphere Fluidics Ltd is commercialising new lab-on-a-chip and picodroplet technology developed at Cambridge University by company founder Professor Chris Abell and Professor Wilhelm Huck. Focussing on storing and retrieving individual picodroplets containing a unique sample, their technology benefits a wide range of applications, including rapid antibody discovery, identification of novel algal strains and the generation of new biocatalysts. For more information please visit www.spherefluidics.com Pico-Surf™ is a trademark of Sphere Fluidics FC-40™ and Novec™ are trademarks of 3M Company MAR-000124_v.A.14 Page 11 of 12
The Dolomite Centre Ltd MAR-000124_v.A.14 Page 12 of 12
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