MAPSYNE: Miniaturized micropipette system combined with high-density microelectrode arrays for automated manipulation of neuronal networks in-vitro

We current MAPSYNE, a miniaturized and automatic system combining a high-density microelectrode array (HD-MEA) and a movable micropipette for learning, monitoring, and perturbing neurons in vitro.

The system includes an all-electrical method to mechanically transfer a glass micropipette in direction of a goal location on the HD-MEA floor, with out the necessity for a microscope. Two strategies of performing blind navigation are employed, (i) stop-measure-go method whereby the pipette strikes for a predefined distance earlier than measuring its location then the method is repeated till the pipette reaches its vacation spot, and (ii) predictive method whereby the pipette is repeatedly tracked and moved.

This automated system could be utilized for unsupervised single-cell manipulation of neurons in a community, corresponding to electroporation and native supply of compounds.

This work reviews the event of an oil immersed-scanning micropipette contact technique, a variant of scanning micropipette contact technique, the place a skinny layer of oil wets the investigated substrate. Oil immersed-scanning micropipette contact technique considerably will increase the droplet stability, permitting for extended mapping and the usage of extremely evaporative saline options no matter ambient humidity ranges.

This systematic mapping method was used to conduct an in depth investigation of localized corrosion going down on the floor of an AA7075-T73 aluminum alloy in a 3.5 wt% NaCl electrolyte, which is often difficult within the standard scanning micropipette contact technique. Maps of corrosion potentials and corrosion currents extracted from potentiodynamic polarization curves confirmed good correlations with the chemical composition of floor options and identified galvanic interactions on the microscale degree.

This demonstrates the viability of oil immersed-scanning micropipette contact technique and opens up the avenue to mechanistic corrosion investigations on the microscale degree utilizing aqueous options which are vulnerable to evaporation underneath non-controlled humidity ranges.

Positioning single cells on a strong floor is an important method for understanding the mobile capabilities and cell-cell interactions in cell tradition assays. We developed a microfluidic chip for depositing single cells in microwells utilizing a easy micropipette operation. Cells had been delivered to microwells by the meniscus movement of liquid interface.

The residue deposits of cells had been redistributed with air injection, and the remoted single cells had been saved in microwells. Different microwell sizes and depths had been studied to guage the trapping chance of cells. Medium substitute and cell viability staining with the remoted single cells had been achieved in microwells. The chip will function a instrument for single-cell patterning in an easy-to-use method.

Phos-tag-based micropipette-tip technique for evaluation of phosphomonoester-type impurities in artificial oligonucleotides

Various chromatographic strategies, mixed with mass spectrometry, have been developed for the evaluation of impurities in oligonucleotide medication, however these strategies have usually been much less centered on attainable phosphomonoester-type compounds. Here, we introduce a easy technique for separating terminally phosphorylated impurities from mother or father oligonucleotides by utilizing a phosphate-affinity micropipette tip (Phos-tag tip).

All steps for the phosphate-affinity separation (binding, washing, and elution) are carried out in aqueous buffers at impartial pH. The total separation protocol requires lower than 30 min per pattern. In sensible examples, we demonstrated that phosphorylated impurities in natural-type and chemically modified oligonucleotides could be effectively separated by the Phos-tag tip technique and subsequently characterised by utilizing ion-pairing reversed-phase liquid chromatography mass spectrometry (IP-RPLC-MS).

Thus, a mixture of the Phos-tag tip technique and IP-RPLC-MS is helpful for characterizing and figuring out phosphomonoester-type impurities in oligonucleotide medication.

Cyclic micropipette aspiration reveals viscoelastic change of a gelatin microgel ready inside a lipid droplet.

Gelatin microgels ready inside lipid droplets have a a lot increased elasticity than that of bulk gels as a consequence of their variations in nanostructure. This nanostructural distinction in gelatin microgels is anticipated to supply the microgels with distinctive viscoelastic properties that differ from the majority gels.

To make clear this speculation, right here we evaluated the frequency dependent viscoelasticity of gelatin gels by growing a cyclic micropipette aspiration. The frequency dependent relationship between storage modulus E’ (reflecting elasticity) and loss modulus E” (reflecting viscosity) was in contrast between the microgels and the majority gels. The microgels have a smaller ratio between elasticity and viscosity E”/E’ than that of the majority gels.

Because the E”/E’ of the majority gels is fixed whatever the focus, the microgel viscoelasticity can’t be achieved for the majority gels with a distinct focus. Theses findings imply that making ready biopolymer gels in droplets is helpful to vary the viscoelasticity by way of nanostructural transition by the interplay with the droplet interface.

Measurements of the unitary hydraulic conductivity of membrane channels, p_f , could also be hampered by difficulties in producing enough portions of purified and reconstituted proteins. Low yield expression, the purely empiric selection of detergents, in addition to protein aggregation and misfolding throughout reconstitution might end in a median of lower than one reconstituted channel per massive unilamellar vesicle.

This limits their applicability for p_f measurements, impartial of whether or not mild scattering or fluorescence quenching of encapsulated dyes is monitored. Here we adopted the micropipette aspiration method as a result of its excellent sensitivity permits resolving p_f values for one order of magnitude smaller protein densities in sphingomyelin and ldl cholesterol wealthy large unilamellar vesicles (GUVs). We derived protein density from depth fluctuations that fluorescently labeled channels within the aspirated GUV induce by diffusing by the diffraction restricted spot.

Fixvo II Micropipette - 500 Capacity -
LA607-1NO	EWC Diagnostics	1 unit	EUR 34.73
Description: Fixvo II Micropipette - 500 Capacity -

Varivol II Micropipette-200 Capacity :
LA613-1NO	EWC Diagnostics	1 unit	EUR 61.99
Description: Varivol II Micropipette-200 Capacity :

Varivol II Micropipette-100 Capacity :
LA615-1NO	EWC Diagnostics	1 unit	EUR 61.99
Description: Varivol II Micropipette-100 Capacity :

Varivol II Micropipette-200 Capacity :
LA618-1NO	EWC Diagnostics	1 unit	EUR 61.99
Description: Varivol II Micropipette-200 Capacity :

Varivol II Micropipette-1000 Capacity
LA614-1NO	EWC Diagnostics	1 unit	EUR 61.99
Description: Varivol II Micropipette-1000 Capacity

Varivol II Micropipette-1000 Capacity
LA616-1NO	EWC Diagnostics	1 unit	EUR 61.99
Description: Varivol II Micropipette-1000 Capacity

Varivol II Micropipette-5000 Capacity
LA619-1NO	EWC Diagnostics	1 unit	EUR 61.99
Description: Varivol II Micropipette-5000 Capacity

Varivol II Micropipette-10 Capacity :
LA611-1NO	EWC Diagnostics	1 unit	EUR 61.99
Description: Varivol II Micropipette-10 Capacity :

Varivol II Micropipette-50 Capacity :
LA612-1NO	EWC Diagnostics	1 unit	EUR 61.99
Description: Varivol II Micropipette-50 Capacity :

Varivol II Micropipette-20 Capacity :
LA617-1NO	EWC Diagnostics	1 unit	EUR 61.99
Description: Varivol II Micropipette-20 Capacity :

Varivol II Micropipette-10000 Capacity :
LA878-1NO	EWC Diagnostics	1 unit	EUR 75.54
Description: Varivol II Micropipette-10000 Capacity :

Fixvo II Micropipette - 5 Capacity - 5
LA599-1NO	EWC Diagnostics	1 unit	EUR 34.73
Description: Fixvo II Micropipette - 5 Capacity - 5

Fixvo II Micropipette - 10 Capacity -
LA600-1NO	EWC Diagnostics	1 unit	EUR 34.73
Description: Fixvo II Micropipette - 10 Capacity -

Micropipette Acura 825 0.1-2 uL Socorex - EACH
DD61104	Scientific Laboratory Supplies	EACH	EUR 488.7

Micropipette Acura 825 2-20 uL Socorex - EACH
DD61107	Scientific Laboratory Supplies	EACH	EUR 450.9

Micropipette Acura 825 20-200 uL Socorex - EACH
DD61110	Scientific Laboratory Supplies	EACH	EUR 450.9

Micropipette Acura 825 100-1000 uL Socorex - EACH
DD61111	Scientific Laboratory Supplies	EACH	EUR 450.9

Micropipette Acura 835 1-10 uL Socorex - EACH
DD61113	Scientific Laboratory Supplies	EACH	EUR 487.35

Pe Filter For Micropipette Protection - PK100
DD061968	Scientific Laboratory Supplies	PK100	EUR 152.55

µPet Autoclavable Micropipette Capacity:
LA955-1NO	EWC Diagnostics	1 unit	EUR 145.85
Description: µPet Autoclavable Micropipette Capacity:

µPet Autoclavable Micropipette Capacity:
LA956-1NO	EWC Diagnostics	1 unit	EUR 145.85
Description: µPet Autoclavable Micropipette Capacity:

µPet Autoclavable Micropipette Capacity:
LA957-1NO	EWC Diagnostics	1 unit	EUR 145.85
Description: µPet Autoclavable Micropipette Capacity:

µPet Autoclavable Micropipette Capacity:
LA958-1NO	EWC Diagnostics	1 unit	EUR 145.85
Description: µPet Autoclavable Micropipette Capacity:

µPet Autoclavable Micropipette Capacity:
LA959-1NO	EWC Diagnostics	1 unit	EUR 145.85
Description: µPet Autoclavable Micropipette Capacity:

µPet Autoclavable Micropipette Capacity:
LA960-1NO	EWC Diagnostics	1 unit	EUR 145.85
Description: µPet Autoclavable Micropipette Capacity:

µPet Autoclavable Micropipette Capacity:
LA961-1NO	EWC Diagnostics	1 unit	EUR 145.85
Description: µPet Autoclavable Micropipette Capacity:

µPet Autoclavable Micropipette Capacity:
LA962-1NO	EWC Diagnostics	1 unit	EUR 145.85
Description: µPet Autoclavable Micropipette Capacity:

µPet Autoclavable Micropipette Capacity:
LA963-1NO	EWC Diagnostics	1 unit	EUR 84.57
Description: µPet Autoclavable Micropipette Capacity:

µPet Autoclavable Micropipette 8-channel
LA964-1NO	EWC Diagnostics	1 unit	EUR 601.15
Description: µPet Autoclavable Micropipette 8-channel

µPet Autoclavable Micropipette 8-channel
LA965-1NO	EWC Diagnostics	1 unit	EUR 601.15
Description: µPet Autoclavable Micropipette 8-channel

µPet Autoclavable Micropipette Capacity:
LA966-1NO	EWC Diagnostics	1 unit	EUR 601.15
Description: µPet Autoclavable Micropipette Capacity:

µPet Autoclavable Micropipette 8-channel
LA967-1NO	EWC Diagnostics	1 unit	EUR 601.15
Description: µPet Autoclavable Micropipette 8-channel

Capillary micropipette Microcaps Drummond 0.5ulger) for pipettes 1 to 100 mL - PK100
DD075071	Scientific Laboratory Supplies	PK100	EUR 49.95

Positive displacement micropipette Acura capillar 846 volume 1 - 2 - 4 - 5 uL - EACH
DD61945	Scientific Laboratory Supplies	EACH	EUR 537.3

Positive displacement micropipette Acura capillar 846 volume 10 - 20 - 30 - 40 - 50 uL - EACH
DD61947	Scientific Laboratory Supplies	EACH	EUR 537.3

Positive displacement micropipette Acura capillar 846 volume 60 - 75 - 80 - 90 - 100 uL - EACH
DD61948	Scientific Laboratory Supplies	EACH	EUR 537.3

Filter Syringe Glass Microfibre - PK500
68061316	Scientific Laboratory Supplies	PK500	EUR 1223.1

WHATMAN(TM) GLASS MICROFIBER FILTRATION
WHA1950009	Scientific Laboratory Supplies	EACH	EUR 282.15

WHATMAN(TM) GLASS MICROFIBER FILTRATION
WHA1950017	Scientific Laboratory Supplies	EACH	EUR 280.17

WHATMAN(TM) GLASS MICROFIBER FILTRATION
WHA1950217	Scientific Laboratory Supplies	EACH	EUR 156.42

WHATMAN GF/F 25.7CM DIAM GLASS MICROFIB
WHA1825257	Scientific Laboratory Supplies	EACH	EUR 418.38

WHATMAN(TM) GLASS MICROFIBER FILTERS GR
WHA18226580	Scientific Laboratory Supplies	EACH	EUR 144.54

WHATMAN(TM) GLASS MICROFIBER FILTERS GR
WHA1822866	Scientific Laboratory Supplies	EACH	EUR 293.04

WHATMAN(TM) GLASS MICROFIBER FILTERS GR
WHA1823007	Scientific Laboratory Supplies	EACH	EUR 108.9

WHATMAN(TM) GLASS MICROFIBER FILTERS GR
WHA1823915	Scientific Laboratory Supplies	EACH	EUR 1101.87

WHATMAN(TM) GLASS MICROFIBER FILTERS GR
WHA1825015	Scientific Laboratory Supplies	EACH	EUR 196.02

WHATMAN(TM) GLASS MICROFIBER FILTERS GR
WHA1825037	Scientific Laboratory Supplies	EACH	EUR 111.63

WHATMAN GLASS MICROFIBER FILTER DIAM. 2
WHA1820025	Scientific Laboratory Supplies	EACH	EUR 26.58

WHATMAN GLASS MICROFIBER FILTER DIAM. 2
WHA1821021	Scientific Laboratory Supplies	EACH	EUR 25.74

WHATMAN GLASS MICROFIBER FILTER DIAM. 2
WHA1821024	Scientific Laboratory Supplies	EACH	EUR 47.52

WHATMAN GLASS MICROFIBER FILTER DIAM. 2
WHA1821025	Scientific Laboratory Supplies	PK100	EUR 51.48

WHATMAN GLASS MICROFIBER FILTER DIAM. 4
WHA1821042	Scientific Laboratory Supplies	PK100	EUR 72.27

WHATMAN GLASS MICROFIBER FILTER DIAM. 4
WHA1821047	Scientific Laboratory Supplies	EACH	EUR 79.2

WHATMAN GLASS MICROFIBER FILTER DIAM. 1
WHA1821150	Scientific Laboratory Supplies	EACH	EUR 96.03

WHATMAN GLASS MICROFIBER FILTER DIAM. 2
WHA1822021	Scientific Laboratory Supplies	PK100	EUR 48.51

WHATMAN GLASS MICROFIBER FILTER DIAM. 2
WHA1822024	Scientific Laboratory Supplies	EACH	EUR 45.54

WHATMAN GLASS MICROFIBER FILTER DIAM. 3
WHA1822037	Scientific Laboratory Supplies	EACH	EUR 51.48

WHATMAN GLASS MICROFIBER FILTER DIAM. 4
WHA1822042	Scientific Laboratory Supplies	EACH	EUR 51.48

WHATMAN GLASS MICROFIBER FILTER DIAM. 1
WHA1822110	Scientific Laboratory Supplies	EACH	EUR 164.34

WHATMAN GLASS MICROFIBER FILTER DIAM. 1
WHA1822150	Scientific Laboratory Supplies	EACH	EUR 361.35

WHATMAN GLASS MICROFIBER FILTER DIAM. 2
WHA1823021	Scientific Laboratory Supplies	PK100	EUR 36.63

WHATMAN GLASS MICROFIBER FILTER DIAM. 2
WHA1823024	Scientific Laboratory Supplies	EACH	EUR 31.68

WHATMAN GLASS MICROFIBER FILTER DIAM. 2
WHA1823025	Scientific Laboratory Supplies	EACH	EUR 39.6

WHATMAN GLASS MICROFIBER FILTER DIAM. 4
WHA1823042	Scientific Laboratory Supplies	EACH	EUR 45.54

WHATMAN GLASS MICROFIBER FILTER DIAM. 7
WHA1823070	Scientific Laboratory Supplies	PK100	EUR 94.05

WHATMAN GLASS MICROFIBER FILTER DIAM. 1
WHA1823110	Scientific Laboratory Supplies	EACH	EUR 40.59

WHATMAN GLASS MICROFIBER FILTER DIAM. 1
WHA1823125	Scientific Laboratory Supplies	EACH	EUR 59.4

WHATMAN GLASS MICROFIBER FILTER DIAM. 1
WHA1823150	Scientific Laboratory Supplies	EACH	EUR 101.97

WHATMAN GLASS MICROFIBER FILTER DIAM. 2
WHA1825021	Scientific Laboratory Supplies	EACH	EUR 66.33

WHATMAN GLASS MICROFIBER FILTER DIAM. 2
WHA1825024	Scientific Laboratory Supplies	EACH	EUR 88.11

WHATMAN GLASS MICROFIBER FILTER DIAM. 2
WHA1825025	Scientific Laboratory Supplies	EACH	EUR 85.8

WHATMAN GLASS MICROFIBER FILTER DIAM. 4
WHA1825042	Scientific Laboratory Supplies	EACH	EUR 122.76

WHATMAN GLASS MICROFIBER FILTER DIAM. 7
WHA1825070	Scientific Laboratory Supplies	EACH	EUR 199.95

WHATMAN GLASS MICROFIBER FILTER DIAM. 1
WHA1825125	Scientific Laboratory Supplies	PK25	EUR 145.53

WHATMAN GLASS MICROFIBER FILTER DIAM. 1
WHA1825150	Scientific Laboratory Supplies	EACH	EUR 189.09

WHATMAN GF/D 25.7CM DIAM GLASS MICROFI - EACH
WHA1823257	Scientific Laboratory Supplies	EACH	EUR 164.7

WHATMAN(TM) GLASS MICROFIBER FILTRATION - EACH
WHA1950002	Scientific Laboratory Supplies	EACH	EUR 261.9

WHATMAN(TM) GLASS MICROFIBER FILTERS GR - EACH
WHA1821914	Scientific Laboratory Supplies	EACH	EUR 396.9

WHATMAN(TM) GLASS MICROFIBER FILTERS GR - PK25
WHA1825293	Scientific Laboratory Supplies	PK25	EUR 756

WHATMAN(TM) GLASS MICROFIBER FILTERS WI - PK200
WHA10421030	Scientific Laboratory Supplies	PK200	EUR 171.45

WHATMAN GLASS MICROFIBER FILTER DIAM. 2 - EACH
WHA1820024	Scientific Laboratory Supplies	EACH	EUR 43.2

WHATMAN GLASS MICROFIBER FILTER DIAM. 4 - EACH
WHA1820042	Scientific Laboratory Supplies	EACH	EUR 49.95

WHATMAN(TM) GLASS MICROFIBER FILTERS GR - PK100
WHA1820240	Scientific Laboratory Supplies	PK100	EUR 278.1

WHATMAN GLASS MICROFIBER FILTER DIAM. 3 - EACH
WHA1821037	Scientific Laboratory Supplies	EACH	EUR 87.75

WHATMAN GLASS MICROFIBER FILTER DIAM. 1 - EACH
WHA1821110	Scientific Laboratory Supplies	EACH	EUR 59.4

WHATMAN GLASS MICROFIBER FILTER DIAM. 5 - EACH
WHA1823055	Scientific Laboratory Supplies	EACH	EUR 81

WHATMAN GLASS MICROFIBER FILTER DIAM. 9 - EACH
WHA1823090	Scientific Laboratory Supplies	EACH	EUR 41.85

WHATMAN GLASS MICROFIBER FILTER DIAM. 1 - PK25
WHA1825110	Scientific Laboratory Supplies	PK25	EUR 163.35

WHATMAN (R) GLASS MICROFIBER MEMBRANE HO - EACH
WHA1960054	Scientific Laboratory Supplies	EACH	EUR 691.2

WHATMAN STANDARD GLASS MICROFIBER EXTRAC - PK25
WHA10371025	Scientific Laboratory Supplies	PK25	EUR 280.8

WHATMAN STANDARD GLASS MICROFIBER EXTRAC - EACH
WHA10371042	Scientific Laboratory Supplies	EACH	EUR 248.4

WHATMAN GLASS MICROFIBER FILTER DIAM. 2 - PK100
WHA1820021	Scientific Laboratory Supplies	PK100	EUR 47.25

WHATMAN GLASS MICROFIBER FILTER DIAM. 4 - PK100
WHA1820047	Scientific Laboratory Supplies	PK100	EUR 74.91

WHATMAN GLASS MICROFIBER FILTER DIAM. 7 - PK100
WHA1821070	Scientific Laboratory Supplies	PK100	EUR 216

WHATMAN GLASS MICROFIBER FILTER DIAM. 5 - PK100
WHA1825055	Scientific Laboratory Supplies	PK100	EUR 271.35

WHATMAN(R) GLASS MICROFIBER FILTERS GRA - PK100
WHA1827025	Scientific Laboratory Supplies	PK100	EUR 32.4

Glass Microfibre Filter Discs Gr SS - 25mm - PK100
SLS4530	Scientific Laboratory Supplies	PK100	EUR 32.4

Glass Microfibre Filter Discs Gr SS - 47mm - PK100
SLS4531	Scientific Laboratory Supplies	PK100	EUR 39.15

Glass Microfibre Filter Discs Gr SS - 55mm - PK100
SLS4532	Scientific Laboratory Supplies	PK100	EUR 51.3

Glass Microfibre Filter Discs Gr SS - 70mm - PK100
SLS4533	Scientific Laboratory Supplies	PK100	EUR 63.45

Glass Microfibre Filter Discs Gr SS - 90mm - PK100
SLS4534	Scientific Laboratory Supplies	PK100	EUR 90.45

Glass Microfibre Filter Discs Gr SS - 110mm - PK100
SLS4535	Scientific Laboratory Supplies	PK100	EUR 113.4

Glass Microfibre Filter Discs Gr SS - 125mm - PK100
SLS4536	Scientific Laboratory Supplies	PK100	EUR 170.1

Glass Microfibre Filter Discs Gr SS - 150mm - PK100
SLS4537	Scientific Laboratory Supplies	PK100	EUR 211.95

Whatman GFB 90mm Membrane Glass Microfibre - PK25
FIL4218	Scientific Laboratory Supplies	PK25	EUR 54

Whatman GFB 110mm Membrane Glass Microfibre - PK25
FIL4220	Scientific Laboratory Supplies	PK25	EUR 63.45

Whatman GFB 125mm Membrane Glass Microfibre - PK25
FIL4222	Scientific Laboratory Supplies	PK25	EUR 82.35

Whatman GFB 150mm Membrane Glass Microfibre - PK25
FIL4226	Scientific Laboratory Supplies	PK25	EUR 125.55

Whatman GFB 185mm Membrane Glass Microfibre - PK25
FIL4228	Scientific Laboratory Supplies	PK25	EUR 157.95

Whatman GFD 90mm Membrane Glass Microfibre - PK25
FIL4420	Scientific Laboratory Supplies	PK25	EUR 44.55

Whatman GFD 110mm Membrane Glass Microfibre - PK25
FIL4422	Scientific Laboratory Supplies	PK25	EUR 59.4

Whatman GFD 125mm Membrane Glass Microfibre - PK25
FIL4424	Scientific Laboratory Supplies	PK25	EUR 76.95

Whatman GFD 150mm Membrane Glass Microfibre - PK25
FIL4426	Scientific Laboratory Supplies	PK25	EUR 132.3

Whatman GFF 90mm Membrane Glass Microfibre - PK25
FIL4516	Scientific Laboratory Supplies	PK25	EUR 91.8

Whatman GFF 110mm Membrane Glass Microfibre - PK25
FIL4518	Scientific Laboratory Supplies	PK25	EUR 125.55

Whatman GFF 125mm Membrane Glass Microfibre - PK25
FIL4520	Scientific Laboratory Supplies	PK25	EUR 152.55

Whatman GFF 150mm Membrane Glass Microfibre - PK25
FIL4522	Scientific Laboratory Supplies	PK25	EUR 222.75

Whatman GFA 55mm Membrane Glass Microfibre - BX100
FIL4116	Scientific Laboratory Supplies	BX100	EUR 59.4

Whatman GFA 70mm Membrane Glass Microfibre - PK100
FIL4120	Scientific Laboratory Supplies	PK100	EUR 74.25

Whatman GFA 90mm Membrane Glass Microfibre - PK100
FIL4122	Scientific Laboratory Supplies	PK100	EUR 105.3

Whatman GFA 125mm Membrane Glass Microfibre - PK100
FIL4126	Scientific Laboratory Supplies	PK100	EUR 174.15

Whatman GFA 150mm Membrane Glass Microfibre - PK100
FIL4128	Scientific Laboratory Supplies	PK100	EUR 211.95

A perfusion system minimized unstirred layers within the instant membrane neighborhood as demonstrated by the distribution of each encapsulated and extravesicular aqueous dyes. p_f amounted to 2.4 ± 0.1 × 10^-13 cm³/s for aquaporin-1 that served as a check case. The new assay paves the way in which for straight monitoring the impact that interplay of aquaporins with different proteins or inhibitors might have on p_f on a single pattern. This article is protected by copyright. All rights reserved.