WO2007136386A2 - Préparation d'échantillons sur puce à base de gouttelettes destinée à la spectrométrie de masse - Google Patents

Préparation d'échantillons sur puce à base de gouttelettes destinée à la spectrométrie de masse Download PDF

Info

Publication number
WO2007136386A2
WO2007136386A2 PCT/US2006/021699 US2006021699W WO2007136386A2 WO 2007136386 A2 WO2007136386 A2 WO 2007136386A2 US 2006021699 W US2006021699 W US 2006021699W WO 2007136386 A2 WO2007136386 A2 WO 2007136386A2
Authority
WO
WIPO (PCT)
Prior art keywords
droplet
plate
electrodes
microfluidic device
droplets
Prior art date
Application number
PCT/US2006/021699
Other languages
English (en)
Other versions
WO2007136386A3 (fr
Inventor
Aaron R. Wheeler
Robin L. Garrell
Chang-Jin Kim
Hyejin Moon
Original Assignee
The Regents Of The University Of California
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by The Regents Of The University Of California filed Critical The Regents Of The University Of California
Publication of WO2007136386A2 publication Critical patent/WO2007136386A2/fr
Publication of WO2007136386A3 publication Critical patent/WO2007136386A3/fr

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
    • G01N33/6848—Methods of protein analysis involving mass spectrometry
    • G01N33/6851—Methods of protein analysis involving laser desorption ionisation mass spectrometry
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502769—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements
    • B01L3/502784—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements specially adapted for droplet or plug flow, e.g. digital microfluidics
    • B01L3/502792—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements specially adapted for droplet or plug flow, e.g. digital microfluidics for moving individual droplets on a plate, e.g. by locally altering surface tension
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0678—Facilitating or initiating evaporation
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0819—Microarrays; Biochips
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/089—Virtual walls for guiding liquids
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0415—Moving fluids with specific forces or mechanical means specific forces electrical forces, e.g. electrokinetic
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0415—Moving fluids with specific forces or mechanical means specific forces electrical forces, e.g. electrokinetic
    • B01L2400/0427—Electrowetting
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/40—Concentrating samples
    • G01N1/4022—Concentrating samples by thermal techniques; Phase changes
    • G01N2001/4027—Concentrating samples by thermal techniques; Phase changes evaporation leaving a concentrated sample

Definitions

  • the present invention relates to the fields of microfluidics, lab-on-a-chip, and micro total analysis system ( ⁇ -TAS), especially droplet-based microfluidics or digital microfluidics.
  • ⁇ -TAS micro total analysis system
  • proteomics has undergone a meteoric rise in popularity, with more than 2000 papers published in the field in 2003.
  • proteomics like genomics, requires methods and instruments capable of collecting, storing, cataloguing, and analyzing vast amounts of information.
  • the technological challenges for proteomics may be even greater than those for genomics, given that an organism has a single genome but may express hundreds of different proteomes, depending on environmental and developmental cues.
  • the development of new methods and instrumentation with the capacity for rapid, high-throughput data collection is crucial for continued progress.
  • MALDI matrix -assisted laser desorption/ionization
  • TOF time-of-flight
  • MALDI-MS Matrix- Assisted Laser Desorption/Ionization-Mass Spectrometry
  • typical proteomics analyses require many steps; a crucial step is mixing the sample with matrix.
  • Repetitive pipetting of reagents onto MALDI targets is time-consuming and can lead to sample loss, dilution, and contamination.
  • High-end commercial instruments utilize robotically controlled deposition, but such instruments are expensive and require careful maintenance.
  • Other methods for high-throughput deposition of sample and matrix include using lithographically patterned targets, microfabricated picoliter droplet delivery devices, or micro fluidic channels. Of these methods, only patterned targets, which facilitate easier spot deposition but do not eliminate pipetting, have gained widespread use.
  • Sample processing for MALDI-MS applications is typically accomplished manually (by pipetting) or in some cases may be partially accomplished by droplet dispensing robots.
  • MALDI-MS is used for many applications, including analysis of proteins, nucleic acids, and synthetic polymers.
  • the present invention relates to droplet-based on-chip sample preparation for mass spectrometry.
  • sample processing is performed in matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) using one or more droplet-based microfluidic devices to dispense, transport, merge, mix, cut, and deliver droplets containing samples and reagents to specified locations on an array.
  • MALDI-MS matrix assisted laser desorption/ionization mass spectrometry
  • reactions may take place in one or more droplets.
  • Substances in droplets may be concentrated by solvent evaporation from the droplets.
  • Droplets may also be used to deposit materials on the device surface by precipitation or evaporation. Droplets may be used to dissolve one or more components from a solid sample on the device, and may be used to transport the dissolved substances.
  • Droplets may also be used to remove solids or particulates from the surface and transport them elsewhere on the device. Samples or substances may be desorbed directly from the device for analysis by mass spectrometry.
  • the mass spectrometry analysis may include peptide mass fingerprinting and database identification.
  • the present invention is a method for cocrystallizing sample and matrix for MALDI-MS.
  • the method utilizes a solution handling technique known as digital ⁇ crofluidics.
  • digital microfluidics droplets are moved over an array of electrodes by means of Electro Wetting-On-Dielectric (EWOD), dielectrophoresis and/or other mechanisms.
  • EWOD Electro Wetting-On-Dielectric
  • the local wettability of a surface is reversibly changed by applying potentials between electrodes buried beneath hydrophobic, dielectric layers.
  • potentials between electrodes buried beneath hydrophobic, dielectric layers By applying a sequence of potentials to adjacent electrodes on an array, liquid droplets can be made to travel across the surface. Droplets may also move in response to an applied field gradient, without any apparent contact angle change, a process known as dielectrophoresis.
  • digital microfluidics any mechanism used to manipulate droplets on an array of electrodes.
  • digital micro fluidics-based devices including single-plate open air devices, parallel-plate devices filled with silicone oil, and parallel-plate open-air devices.
  • Digital microfluidics-based devices are reconfigurable and can handle neutral and charged analytes, particulates, proteins, cells and microorganisms.
  • the present invention also discloses a method to realize digital microfluidics actuation across a two-dimensional plane (rather than simply across one or two rows of electrodes) and to use this technique to create a fully portable micro fluidic device.
  • Digital microfluidics is well-suited to MALDI, as both techniques rely on array geometries, which stands in contrast to channel-based microfluidic devices for MALDI-MS, which require rastering or complex networks of holes to mate with MALDI-MS targets.
  • a method in accordance with the present invention comprises performing sample processing using at least one droplet-based microfluidic device capable of manipulating droplets containing samples, placing the droplets at specified locations on an array; and moving the droplets within the array.
  • Such a method optionally includes the droplet-based microfluidic devices being formed in an array geometry, applying a sequence of electrical signals to enable transport of droplets across the droplet-based microfluidic device surface, the sequence of electrical signals being applied to a pattern of electrodes buried beneath a dielectric layer on the droplet-based microfluidic device surface, the sequence of electrical signals dividing the droplet, and the droplets comprising a homogeneous liquid, emulsion or suspension.
  • the method further optionally comprises placing a droplet containing a reagent on the array, and moving the droplet containing the reagent within the array, the droplets comprising solutions in which cells or particles are suspended, the samples reacting with chemical agents in the droplets, the samples being acted on by catalysts in the droplets, the samples being concentrated by evaporating liquid from the droplet, the samples being precipitated from the droplet, the samples being crystallized by evaporating liquid from the droplet, the droplets being used to dissolve the samples, the samples being purified by selective precipitation, the droplet-based microfluidics device being used with a mass spectrometer, and a plurality of samples being processed in parallel.
  • a device in accordance with the present invention comprises a first plate comprising an array of first electrodes, a second plate, comprising at least a second electrode, wherein the first plate and the second plate are spaced apart such that a droplet can travel between the first plate and the second plate, a first layer of one or more materials, covering the array of first electrodes, and a second layer of one or more materials, covering the at least second electrode, wherein application of electrical signals between selective electrodes within the array of first electrodes and the at least one second electrode moves the droplet between the top plate and the bottom plate.
  • Such a device further optionally includes the first layer of material being Teflon or other coating material, a first droplet being moved between the first plate and the second plate, and dried between the first plate and the second plate, a second droplet being moved between the first plate and the second plate to where the first droplet was dried, the second droplet containing a reagent.
  • the dried spot is then analyzed in situ by MALDI-MS. In a two-plate device, this entails removing the top plate.
  • Such a device further optionally includes devices with a single plate with an array of electrodes.
  • droplets are manipulated on the array by applying potentials between electrodes on a single plane, with no need for a top plate.
  • the droplet is then analyzed in situ by MALDI-MS.
  • FIGS. l(a)-(f) illustrate a matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) application
  • FIGS. 2-3 illustrate digital micro fluidics devices in accordance with the present invention
  • FIGS. 4A-4B illustrate droplet movement experiment on an digital microfluidics device
  • FIG. 5 shows images and mass spectra of insulin cocystallized with DHB, FA, and SA;
  • FIGS. 6A-6C illusrate sample MALDI spectra of insulin-DHB created with different techniques in accordance with the present invention
  • FIGS. 7A- 7C illustrate other proteins and peptides as analyzed with digital microfiuidics-MALDI in accordance with the present invention
  • FIGS. 8A-8B illustrate residual nonspecific adsorption for various sizes of insulin spots in accordance with the present invention
  • FIG. 9 illustrates a process chart showing typical steps used in practicing the invention
  • FIGS. 10A-D illustrate a digital microfluidics-driven reduction of insulin into the constitutive peptides (insulin A and insulin B) followed by tryptic digestion.
  • the present invention enables automated, integrated, reconfigurable, high- throughput sample processing for matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) applications.
  • Sample processing for MALDI-MS is accomplished manually (by pipetting) or in some cases may be partially accomplished by droplet dispensing robots.
  • the present invention relies on droplet-based microfluidics (also known as "digital microfluidics") to create, dispense, transport, merge, mix, cut, and deliver droplets containing samples and reagents to specified locations on an array.
  • Droplet-based microfluidics is described in detail in the publications attached hereto as appendices. Creating a droplet can mean dispensing a droplet from a reservoir, ejecting a droplet from a channel, or dividing a larger droplet.
  • droplet-based microfludics a sequence of potentials is applied to adjacent electrodes buried beneath dielectric layers.
  • a potential is applied across electrodes in the device, two phenomena may be observed: (1) a droplet may move towards the biased electrode, and (2) the contact angle between the droplet and device surface may decrease because of a change in the local wettability of the surface. A change in surface wettability is not required for droplet movement.
  • Droplet-based microfluidic devices are formed in an array geometry, which makes it an attractive technology for use with MALDI-MS (which is typically used to analyze arrays of processed samples).
  • the present invention is the first to use droplet-based microfluidics for on-chip sample preparation on an array for mass spectrometry.
  • the present invention makes use of technology for manipulating droplets of homogeneous liquids, as well as liquid droplets that contain suspended liquid droplets (emulsions) or suspended solids (suspensions), cells or microorganisms, by droplet- based microfluidics, which is described in detail in the publications attached hereto as appendices.
  • a homogeneous liquid may be a pure liquid, or a liquid in which one or more components have been dissolved.
  • Droplet manipulations may include any or all of the following functions: droplet generation or dispensing, movement, merging, dividing or cutting, joining, mixing, and concentrating or drying by evaporation. Droplets may also be manipulated so as to dissolve or suspend solids.
  • the droplets may consist of pure liquids, solvents, solutions, or suspensions. Soluble substances or suspended particles in the droplets may be reagents, polymers or other agents such as surfactants that modify the physical properties of the droplets, samples (analytes), labeling agents (molecular or particulate) and/or catalysts, collectively called chemical agents herein.
  • the present invention required the development of specific device designs and utilization parameters to establish feasibility of manipulating samples and reagents for MALDI-MS.
  • MALDI-MS is used for many applications, including analysis of proteins, nucleic acids, and synthetic polymers. It is in the former capacity, the analysis of proteins, that this technique has become an extremely important tool used in virtually every biochemistry laboratory in the world. Proteomics, the study of all proteins expressed in a given sample, is an important field in chemistry and biology, with applications for both clinical diagnostics and basic science. The capacity to streamline the processing required for proteomics and other MALDI-MS applications will be widely desired in both industry and academia.
  • the method has been demonstrated to be useful for the . function of mixing and crystallizing protein samples and MALDI matrices.
  • the method has also been demonstrated to be useful for the purification step, and for the combining of reagents or catalysts, such as reducing agents, alkylating agents or enzymes, with samples to be analyzed and performing those reactions on the device.
  • reagents or catalysts such as reducing agents, alkylating agents or enzymes
  • FIGS. l(a)-(f) A MALDI- MS proteomics analysis is described in FIGS. l(a)-(f).
  • FIG. l(a) illustrates the step of 2-D gel electrophoresis
  • FIG. l(b) illustrates the step of excising separated protein spots
  • FIG. l(c) illustrates the step of processing the samples, which may include any or all of the following: sample purification, disulfide reduction, proteolytic digestion and peptide recovery
  • FIG. l(d) illustrates the step of crystallizing the sample with a matrix, wherein a matrix can be a substance that co-deposits with the sample
  • FIG. l(e) illustrates the step of collecting peptide mass fingerprints using MALDI
  • FIG. l(e) illustrates the step of identifying the protein by searching one or more proteomic databases.
  • FIGS. l(c) and l(d) typically require between several hours to days to accomplish, involving tens-hundreds of pipetting steps.
  • Expensive, high-end MALDI-MS instruments are equipped with robots that can accomplish the step of FIG. l(d), but the processes in the step of FIG. l(c) remain a bottleneck in virtually all MALDI-MS proteomics methods.
  • droplet-based microfluidics is used to integrate and automate the steps of FIGS. l(c) and l(d).
  • the steps of FIGS. l(a) and l(b) may also be incorporated by integrating micro fluidic separation columns onto a single high-throughput proteomics analysis platform. This would distill a process that requires many hours or days of laboratory time into an automated, high-throughput, programmable, and reconfigurable method that will require only minutes of laboratory time.
  • Stock solutions of analytes including bovine insulin (100 ⁇ M), bovine insulin chain B (40 ⁇ M), horse heart cytochrome c (14.5 ⁇ M), and horse skeletal myoglobin (59 ⁇ M), were prepared in deionized (DI) water or with 0.2% trifluoroacetic acid (TFA). Stock solutions were kept frozen; working solutions were diluted and used within 1 day. Working solutions of matrixes, including 2,5-dihydroxybenzoic acid (DHB), ferulic acid (FA), and sinapinic acid (SA), were prepared in DI water containing TFA and acetonitrile and were used within 1 day.
  • DI deionized
  • TFA horse heart cytochrome c
  • SA sinapinic acid
  • Teflon- AF 1600 resin was purchased from DuPont (Wilmington, DE). Working solutions of 6% (w/v) were formed in Fluorinert FC-40 solvent; solutions were used as made or diluted (v/v with FC-40).
  • FIGS. 2-3 illustrate a digital microfluidic device in accordance with the present invention.
  • device 200 is formed each device was formed from a bottom plate 202 having individually addressable electrodes 204-212 and a top plate 214 with one contiguous electrode 216.
  • Droplet 218 is shown between bottom plate 202 and top plate 214.
  • Bottom plate 202 is formed from quartz wafers coated with a 3500- A layer of phosphorus-doped polysilicon.
  • Polysilicon electrodes 204-212 are patterned using standard photolithographic techniques and reactive ion etching. A typical formation of the electrodes 204-212 is to grow a thermal oxide (1500 A) on the polysilicon of bottom plate 202 in an oxidation furnace. Holes through the oxide to the electrical contacts were formed with photolithography and wet etching with buffered hydrofluoric acid. The devices were then primed with hexamethyldisilazane vapor and spin-coated (2000 rpm, 60 s) with 5% Teflon-AF layer 220.
  • the devices were postbaked on a hot plate (160 0 C, 10 min) and in a furnace (330 0 C, 30 min) to form a uniform 7500- A layer 220 of Teflon-AF.
  • Layer 220 cam be made from any polymer to achieve the desired surface hydrophobicity or hydrophilicity, or wettability. Additional layers 221 of dielectric or other material can be used to control the electron flow between electrodes 204-212 and any droplets that are placed on layer 220.
  • the top plate 214 is a glass piece with the electrode 216 being formed from indium-tin oxide (ITO).
  • ITO indium-tin oxide
  • a 150 A layer 222 of Teflon- AF was spin-coated (0.5%, processed as above) onto the ITO-coated top plate 214.
  • Layer 222 can be made from any polymer to achieve the desired surface hydrophobicity or hydrophilicity, and does not have to be the same material as layer 220.
  • the two plates 202 and 214 are joined with spacers 224 (at approximately 300 ⁇ m apart), which can be formed from three pieces of double-sided tape or other materials. Other spacings between plates 202 and 214 are possible within the scope of the present invention.
  • FIG. 3 illustrates a top view of plate 202 in a typical digital microfluidics pattern, which comprises sixteen 1-mm 2 electrodes 204-212 (having a typical 4- ⁇ m gap between electrodes) where each electrode 204-212 is connected to an electrical contact pad 226.
  • Aqueous droplets 218 (0.5 ⁇ L) are sandwiched between the two plates 202 and 214 and are moved by applying ac potentials (1 kHz, 75 Vrms) between the electrode 216 in the top plate 214 and successive electrodes 204-212 in the bottom plate 202.
  • ac potentials (1 kHz, 75 Vrms
  • digital microfluidics devices 200 were stored in a chamber under house vacuum; 0.5- ⁇ L droplets dried in 1 -2 min. Matrix and sample cocrystals were imaged by light microscopy. Typically, several spots were deposited on each digital microfluidics device 200.
  • the bottom plate of the digital microfluidics device 200 was affixed with double-sided tape into a 1-mm-deep milled-out groove on a standard stainless steel MALDI target. A mass spectrometer is then used to collect MALDI-MS data.
  • acetonitrile is often used to increase the solubility of matrixes for MALDI-MS.
  • the liquids include solvents having a wide range of polarities, and include ionic liquids, chlorinated solvents, alcohols, carboxylic acids, aldehydes, amides, sulfoxides, ethers, heterocycles and nitriles, but are not limited to these listed liquids.
  • FIGS. 4A-4B illustrate droplet movement experiment on an digital microfluidics device.
  • a droplet 400 of insulin was moved to a designated electrode 402 as shown in FIG. 4a.
  • the droplet 400 was then allowed to dry on electrode 402.
  • a droplet 404 of FA was moved to the electrode 402 on top of the droplet 400 of insulin.
  • Droplet 404 was then allowed to dry.
  • dried spot FIG. 4b; and (4) the droplet was allowed to dry. Droplets were routinely driven on and between each line of electrodes on each device.
  • FIGS. 5A-5D show images and mass spectra of insulin cocystallized with DHB, FA, and SA.
  • MALDI-MS was used to analyze spots of protein and matrix prepared by digital microfluidics.
  • SA spots 504 were deposited manually followed by digital microfluidics- driven movement and drying of an insulin droplet. This result demonstrates that if matrixes that are not water soluble are desirable, the technique of pre-coating a high- throughput target with matrix could be used for digital microfluidics-MALDI devices.
  • five spots 506 were prepared by depositing insulin and then DHB on an digital microfluidics device and on a stainless steel target (FIG. 5D).
  • the two kinds of spectra had similar S/N ( 43.1 and 68.4 for conventional MALDI and digital microfluidics-MALDI, respectively).
  • Cocrystallization of sample and matrix for MALDI-MS is process-driven, with many recipes and variants to choose from.
  • FIGS. 6A-6C illusrate sample MALDI spectra of insulin-DHB created with different techniques in accordance with the present invention.
  • Spots formed with the sample first technique were prepared as depicted in FIG. 4.
  • Spots formed with the dried drop technique were prepared by using digital microfluidics to merge a droplet of insulin and DHB and allowing the combined droplet to dry. Prior to drying, the droplet was moved back and forth between electrodes several times, which has been shown to increase mixing efficiency.
  • Spots formed with the sandwich technique were prepared by using digital microfluidics to deposit a droplet of DHB, insulin, and then DHB again.
  • the signal-to-noise ratios 600-604 were similar for spectra formed by each technique, with the sample first and sandwich techniques giving a slightly narrower analyte peak.
  • the compatibility of digital microfluidics- MALDI-MS with common matrix/sample preparation recipes demonstrates that digital microfluidics should be useful for many applications of MALDIMS.
  • FIGS. 7A-7C In addition to insulin (FIGS. 5 and 6), several other proteins and peptides were analyzed with digital microfluidics-MALDI, as shown in FIGS. 7A-7C. Samples with a wide range of molecular weights were probed, including insulin chain B (3495), cytochrome c (12 400), and myoglobin (16 900).
  • FIGS. 8A-8B MALDI-MS proved to be a convenient tool to probe this phenomenon.
  • Two sets of spectra are shown, formed from droplets containing 1.75 or 0.175 ⁇ M insulin (FIGS. 8a and 8b, respectively). For each concentration, a droplet of insulin was moved across a series of electrodes to a designated point and dried. DHB droplets were then moved and deposited some to the dried spot of insulin (main panels of FIGS.
  • the present invention may be a fully integrated component of a mass spectrometer, as well as a standalone component.
  • the present invention works as a sample preparation device that processes samples in parallel rather than sequentially.
  • the present invention also allows processing of each sample in an individualized way. Differences in sample processing may include the number of steps, the type of steps and reagents used for each step, the concentrations of samples and reagents, and the time allowed for each step.
  • FIGS. 10A-D illustrate a digital microfluidics-driven reduction of insulin into the constitutive peptides (insulin A and insulin B) followed by tryptic digestion.
  • a 0.5 ⁇ L droplet of insulin (5 ⁇ M in 3:1 acetonitrile:DI water) was merged with a 0.6 ⁇ L droplet containing the reducing reagent, TCEP (500 ⁇ g/mL in DI water).
  • the combined droplet was actively mixed ( ⁇ 5 min, RT) and then merged with a 1 ⁇ L droplet containing trypsin (0.5 ⁇ M in DI water).
  • the combined droplet was again actively mixed (-10 min, RT) and then dried.
  • Matrix was added, and a MALDI mass spectrum was collected, which is shown in FIG. 1OB.
  • cytochrome c and ubiquitin were digested by merging 0.5 ⁇ L droplets of analyte (2 ⁇ M in 3:1 acetonitrile:DI water) and trypsin (0.5 ⁇ M in DI water), and actively mixing ( ⁇ 10 min, RT); the combined droplet was then dried and exposed to matrix.
  • MALDI mass spectra were collected (FIGS. 1OC and 10D). Asterisks denote peptides identified by the proteomic database search engine, Mascot. For each of these spectra, the Mowse scores (61 and 95 respectively) identifications were significant (p ⁇ 0.05). Sequence coverages were 40 and 51%. Although the analytes in Fig. 10A- D were not completely digested, these preliminary data demonstrate that digital micro fluidics can be used for rapid processing of proteomic samples.
  • FIG. 1OA is a video sequence depicting reduction and digestion of a droplet of insulin
  • FIG. 1OB is a figure of the spectrum of a processed insulin spot.
  • FIGS. 1OC and 1OD show tryptic digests of cytochrome c and ubiquitin (processed by digital microfluidics). Each reaction represented was allowed to react for no more than 30 min, at room temperature.
  • FIG. 9 illustrates a process chart showing typical steps used in practicing the invention.
  • Box 900 illustrates performing sample processing using at least one droplet- based micro fluidic device capable of manipulating droplets containing samples;
  • Box 902 illustrates placing the droplets at specified locations on an array within the droplet-based microfluidic device.
  • Box 904 illustrates moving the droplets within the array.
  • a method in accordance with the present invention comprises performing sample processing using at least one droplet-based microfluidic device capable of manipulating droplets containing samples, placing the droplets at specified locations on an array, and moving the droplets within the array.
  • Such a method optionally includes the droplet-based microfluidic devices being formed in an array geometry, applying a sequence of electrical signals to enable transport of droplets across the droplet-based microfluidic device surface, the sequence of electrical signals being applied to a pattern of electrodes buried beneath a dielectric layer on the droplet-based microfluidic device surface, the sequence of electrical signals dividing the droplet, the droplets comprising a homogeneous liquid, and the sample having particles or cells suspended in the droplet.
  • the method further optionally comprises placing a droplet containing a reagent on the array, and moving the droplet containing the reagent within the array, the droplets comprising solutions in which cells are suspended, the samples being concentrated by evaporating liquid from the droplet, the samples being crystallized by evaporating liquid from the droplet, the droplets being used to dissolve the samples, the samples being purified by selective precipitation, the droplet-based microfluidics device being integrated with a mass spectrometer, and a plurality of samples being processed in parallel.
  • a device in accordance with the present invention comprises a first plate comprising an array of first electrodes, a second plate, comprising at least a second electrode, wherein the first plate and the second plate are spaced apart such that a droplet can travel between the first plate and the second plate, a first layer of material, covering the array of first electrodes, and a second layer of material, covering the at least second electrode, wherein application of electrical signals between selective electrodes within the array of first electrodes and the at least one second electrode moves the droplet between the top plate and the bottom plate.
  • Such a device further optionally includes the first layer of material being Teflon or other coating materials, a first droplet being moved between the first plate and the second plate, and dried between the first plate and the second plate, a second droplet being moved between the first plate and the second plate to where the first droplet was dried, the second droplet containing a reagent, and the droplet being analyzed by a mass spectrometer.
  • the top plate must be removed prior to analysis by mass spectrometry, for one-plate devices (i.e., with no top), the device can be inserted into the spectrometer immediately after processing. It is intended that the scope of the invention be limited not by this detailed description, but rather by the claims appended hereto.

Abstract

La présente invention concerne une préparation d'échantillons sur puce à base de gouttelettes destinée à la spectrométrie de masse. Plus spécifiquement, le traitement de l'échantillon est effectué au moyen d'une spectrométrie de masse par désorption/ionisation laser assistée par matrice (MALDI-MS) utilisant un ou plusieurs dispositifs microfluidiques à base de gouttelettes afin de manipuler des gouttelettes contenant des échantillons et des réactifs et de les placer à des emplacements spécifiques sur un réseau.
PCT/US2006/021699 2005-06-06 2006-06-06 Préparation d'échantillons sur puce à base de gouttelettes destinée à la spectrométrie de masse WO2007136386A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US68766205P 2005-06-06 2005-06-06
US60/687,662 2005-06-06

Publications (2)

Publication Number Publication Date
WO2007136386A2 true WO2007136386A2 (fr) 2007-11-29
WO2007136386A3 WO2007136386A3 (fr) 2008-11-27

Family

ID=38723728

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2006/021699 WO2007136386A2 (fr) 2005-06-06 2006-06-06 Préparation d'échantillons sur puce à base de gouttelettes destinée à la spectrométrie de masse

Country Status (1)

Country Link
WO (1) WO2007136386A2 (fr)

Cited By (38)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009100516A1 (fr) * 2008-02-11 2009-08-20 Wheeler Aaron R Culture cellulaire et dosages cellulaires utilisant des systèmes microfluidiques numériques
WO2010069977A1 (fr) * 2008-12-17 2010-06-24 Tecan Trading Ag Système et instrument de traitement d'échantillons biologiques et de manipulation de liquides contenant des échantillons biologiques
US8053239B2 (en) 2008-10-08 2011-11-08 The Governing Council Of The University Of Toronto Digital microfluidic method for protein extraction by precipitation from heterogeneous mixtures
WO2012001421A1 (fr) 2010-07-01 2012-01-05 Cambridge Enterprise Limited Spectrométrie de masse à ionisation
FR2967148A1 (fr) * 2010-11-10 2012-05-11 Commissariat Energie Atomique Procede d'evaporation controlee d'une goutte liquide dans un dispositif microfluidique.
US8187864B2 (en) 2008-10-01 2012-05-29 The Governing Council Of The University Of Toronto Exchangeable sheets pre-loaded with reagent depots for digital microfluidics
US8202736B2 (en) 2009-02-26 2012-06-19 The Governing Council Of The University Of Toronto Method of hormone extraction using digital microfluidics
WO2014183118A1 (fr) * 2013-05-10 2014-11-13 The Regents Of The University Of California Plateforme microfluidique numérique pour créer, conserver et analyser des spérules de cellule en trois dimensions
US9039973B2 (en) 2008-10-10 2015-05-26 The Governing Council Of The University Of Toronto Hybrid digital and channel microfluidic devices and methods of use thereof
US9097662B2 (en) 2006-04-18 2015-08-04 Advanced Liquid Logic, Inc. Droplet-based particle sorting
WO2016161402A1 (fr) 2015-04-03 2016-10-06 Abbott Laboratories Dispositifs et procédés d'analyse d'échantillon
US9476811B2 (en) 2010-10-01 2016-10-25 The Governing Council Of The University Of Toronto Digital microfluidic devices and methods incorporating a solid phase
WO2017004463A1 (fr) 2015-07-01 2017-01-05 Abbott Laboratories Dispositifs et procédés d'analyse d'échantillon
CN107267383A (zh) * 2013-12-18 2017-10-20 东洋制罐集团控股株式会社 培养容器的制造方法和固相化装置
US9851365B2 (en) 2009-02-26 2017-12-26 The Governing Council Of The University Of Toronto Digital microfluidic liquid-liquid extraction device and method of use thereof
WO2018067878A1 (fr) 2016-10-05 2018-04-12 Abbott Laboratories Dispositifs et procédés d'analyse d'échantillon
CN109420531A (zh) * 2017-08-31 2019-03-05 厦门大学 微流控芯片及其试剂的储存和释放方法
US10232374B2 (en) 2010-05-05 2019-03-19 Miroculus Inc. Method of processing dried samples using digital microfluidic device
US10464067B2 (en) 2015-06-05 2019-11-05 Miroculus Inc. Air-matrix digital microfluidics apparatuses and methods for limiting evaporation and surface fouling
WO2019213583A1 (fr) 2018-05-04 2019-11-07 Abbott Laboratories Procédé d'échantillonnage séquentiel pour améliorer la sensibilité et la cinétique de dosage immunologique d'échantillons de faible volume
US10596572B2 (en) 2016-08-22 2020-03-24 Miroculus Inc. Feedback system for parallel droplet control in a digital microfluidic device
CN110918145A (zh) * 2019-12-19 2020-03-27 上海天马微电子有限公司 微流控面板及其驱动方法
CN110918146A (zh) * 2019-12-19 2020-03-27 上海天马微电子有限公司 微流控面板
CN111054455A (zh) * 2019-12-26 2020-04-24 上海天马微电子有限公司 微流控芯片及其驱动方法
US10695762B2 (en) 2015-06-05 2020-06-30 Miroculus Inc. Evaporation management in digital microfluidic devices
WO2020140071A1 (fr) 2018-12-28 2020-07-02 Abbott Laboratories Détection directe de molécules simples sur des microparticules
WO2021026403A1 (fr) 2019-08-07 2021-02-11 Abbott Laboratories Méthodes de détection de substances interférant avec une analyse et d'augmentation de l'échelle dynamique
WO2021026402A1 (fr) 2019-08-07 2021-02-11 Abbott Laboratories Composés chimioluminescents pour le multiplexage
US20210205813A1 (en) * 2019-12-23 2021-07-08 National Technology & Engineering Solutions Of Sandia, Llc Contactless liquid loading to microfluidic devices
US11253860B2 (en) 2016-12-28 2022-02-22 Miroculus Inc. Digital microfluidic devices and methods
US11311882B2 (en) 2017-09-01 2022-04-26 Miroculus Inc. Digital microfluidics devices and methods of using them
US11413617B2 (en) 2017-07-24 2022-08-16 Miroculus Inc. Digital microfluidics systems and methods with integrated plasma collection device
US11524298B2 (en) 2019-07-25 2022-12-13 Miroculus Inc. Digital microfluidics devices and methods of use thereof
US11623219B2 (en) 2017-04-04 2023-04-11 Miroculus Inc. Digital microfluidics apparatuses and methods for manipulating and processing encapsulated droplets
US11633738B2 (en) 2015-04-03 2023-04-25 Abbott Laboratories Devices and methods for sample analysis
WO2023122232A1 (fr) * 2021-12-23 2023-06-29 Amgen Inc. Procédés de détection d'une protéine dans un échantillon dans un dispositif fluidique à l'aide d'une spectrométrie de masse
US11738345B2 (en) 2019-04-08 2023-08-29 Miroculus Inc. Multi-cartridge digital microfluidics apparatuses and methods of use
US11772093B2 (en) 2022-01-12 2023-10-03 Miroculus Inc. Methods of mechanical microfluidic manipulation

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040058450A1 (en) * 2002-09-24 2004-03-25 Pamula Vamsee K. Methods and apparatus for manipulating droplets by electrowetting-based techniques

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040058450A1 (en) * 2002-09-24 2004-03-25 Pamula Vamsee K. Methods and apparatus for manipulating droplets by electrowetting-based techniques

Cited By (63)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9097662B2 (en) 2006-04-18 2015-08-04 Advanced Liquid Logic, Inc. Droplet-based particle sorting
US8367370B2 (en) 2008-02-11 2013-02-05 Wheeler Aaron R Droplet-based cell culture and cell assays using digital microfluidics
WO2009100516A1 (fr) * 2008-02-11 2009-08-20 Wheeler Aaron R Culture cellulaire et dosages cellulaires utilisant des systèmes microfluidiques numériques
US9249443B2 (en) 2008-02-11 2016-02-02 The Governing Council Of The University Of Toronto Cell culture and cell assays using digital microfluidics
US8993348B2 (en) 2008-10-01 2015-03-31 The Governing Council Of The University Of Toronto Exchangeable carriers pre-loaded with reagent depots for digital microfluidics
US8187864B2 (en) 2008-10-01 2012-05-29 The Governing Council Of The University Of Toronto Exchangeable sheets pre-loaded with reagent depots for digital microfluidics
US8053239B2 (en) 2008-10-08 2011-11-08 The Governing Council Of The University Of Toronto Digital microfluidic method for protein extraction by precipitation from heterogeneous mixtures
US9039973B2 (en) 2008-10-10 2015-05-26 The Governing Council Of The University Of Toronto Hybrid digital and channel microfluidic devices and methods of use thereof
US10632467B2 (en) 2008-12-17 2020-04-28 Tecan Trading Ag Cartridge, kit and method for manipulating liquids having biological samples
US9545631B2 (en) 2008-12-17 2017-01-17 Tecan Trading Ag Cartridge, kit and method for processing biological samples and manipulating liquids having biological samples
EP2602333A1 (fr) * 2008-12-17 2013-06-12 Tecan Trading AG Systeme et instrument de traitement d'echantillons biologiques et de manipulation de liquides contenant des echantillons biologiques
US8882982B2 (en) 2008-12-17 2014-11-11 Tecan Trading Ag System and instrument for processing biological samples and manipulating liquids having biological samples
US10357773B2 (en) 2008-12-17 2019-07-23 Tecan Trading Ag Cartridge, kit and method for manipulating liquids having biological samples
US8936708B2 (en) 2008-12-17 2015-01-20 Tecan Trading Ag Manipulating the size of liquid droplets in digital microfluidics
CH700127A1 (de) * 2008-12-17 2010-06-30 Tecan Trading Ag System und Vorrichtung zur Aufarbeitung biologischer Proben und zur Manipulation von Flüssigkeiten mit biologischen Proben.
WO2010069977A1 (fr) * 2008-12-17 2010-06-24 Tecan Trading Ag Système et instrument de traitement d'échantillons biologiques et de manipulation de liquides contenant des échantillons biologiques
US8202736B2 (en) 2009-02-26 2012-06-19 The Governing Council Of The University Of Toronto Method of hormone extraction using digital microfluidics
US9851365B2 (en) 2009-02-26 2017-12-26 The Governing Council Of The University Of Toronto Digital microfluidic liquid-liquid extraction device and method of use thereof
US10232374B2 (en) 2010-05-05 2019-03-19 Miroculus Inc. Method of processing dried samples using digital microfluidic device
US11000850B2 (en) 2010-05-05 2021-05-11 The Governing Council Of The University Of Toronto Method of processing dried samples using digital microfluidic device
US9490111B2 (en) 2010-07-01 2016-11-08 Cambridge Enterprise Limited Microdroplet ionisation mass spectrometry
WO2012001421A1 (fr) 2010-07-01 2012-01-05 Cambridge Enterprise Limited Spectrométrie de masse à ionisation
US9476811B2 (en) 2010-10-01 2016-10-25 The Governing Council Of The University Of Toronto Digital microfluidic devices and methods incorporating a solid phase
EP2453220A1 (fr) * 2010-11-10 2012-05-16 Commissariat à l'Énergie Atomique et aux Énergies Alternatives Procédé d'évaporation contrôlée d'une goutte liquide dans un dispositif microfluidique
FR2967148A1 (fr) * 2010-11-10 2012-05-11 Commissariat Energie Atomique Procede d'evaporation controlee d'une goutte liquide dans un dispositif microfluidique.
WO2014183118A1 (fr) * 2013-05-10 2014-11-13 The Regents Of The University Of California Plateforme microfluidique numérique pour créer, conserver et analyser des spérules de cellule en trois dimensions
US11884907B2 (en) 2013-12-18 2024-01-30 Toyo Seikan Group Holdings, Ltd. Culture container, method for culturing lymphocytes, culture-container production method, and solid-phasing apparatus
CN107267383A (zh) * 2013-12-18 2017-10-20 东洋制罐集团控股株式会社 培养容器的制造方法和固相化装置
WO2016161402A1 (fr) 2015-04-03 2016-10-06 Abbott Laboratories Dispositifs et procédés d'analyse d'échantillon
EP3839507A1 (fr) 2015-04-03 2021-06-23 Abbott Laboratories Dispositifs et procédés d'analyse d'échantillons
US11633738B2 (en) 2015-04-03 2023-04-25 Abbott Laboratories Devices and methods for sample analysis
US11022598B2 (en) 2015-04-03 2021-06-01 Abbott Laboratories Devices and methods for sample analysis
US10464067B2 (en) 2015-06-05 2019-11-05 Miroculus Inc. Air-matrix digital microfluidics apparatuses and methods for limiting evaporation and surface fouling
US11471888B2 (en) 2015-06-05 2022-10-18 Miroculus Inc. Evaporation management in digital microfluidic devices
US11097276B2 (en) 2015-06-05 2021-08-24 mirOculus, Inc. Air-matrix digital microfluidics apparatuses and methods for limiting evaporation and surface fouling
US11890617B2 (en) 2015-06-05 2024-02-06 Miroculus Inc. Evaporation management in digital microfluidic devices
US10695762B2 (en) 2015-06-05 2020-06-30 Miroculus Inc. Evaporation management in digital microfluidic devices
US11944974B2 (en) 2015-06-05 2024-04-02 Miroculus Inc. Air-matrix digital microfluidics apparatuses and methods for limiting evaporation and surface fouling
WO2017004463A1 (fr) 2015-07-01 2017-01-05 Abbott Laboratories Dispositifs et procédés d'analyse d'échantillon
US10596572B2 (en) 2016-08-22 2020-03-24 Miroculus Inc. Feedback system for parallel droplet control in a digital microfluidic device
US11298700B2 (en) 2016-08-22 2022-04-12 Miroculus Inc. Feedback system for parallel droplet control in a digital microfluidic device
US11016053B2 (en) 2016-10-05 2021-05-25 Abbott Laboratories Devices and methods for sample analysis
WO2018067878A1 (fr) 2016-10-05 2018-04-12 Abbott Laboratories Dispositifs et procédés d'analyse d'échantillon
US11253860B2 (en) 2016-12-28 2022-02-22 Miroculus Inc. Digital microfluidic devices and methods
US11833516B2 (en) 2016-12-28 2023-12-05 Miroculus Inc. Digital microfluidic devices and methods
US11623219B2 (en) 2017-04-04 2023-04-11 Miroculus Inc. Digital microfluidics apparatuses and methods for manipulating and processing encapsulated droplets
US11413617B2 (en) 2017-07-24 2022-08-16 Miroculus Inc. Digital microfluidics systems and methods with integrated plasma collection device
US11857969B2 (en) 2017-07-24 2024-01-02 Miroculus Inc. Digital microfluidics systems and methods with integrated plasma collection device
CN109420531A (zh) * 2017-08-31 2019-03-05 厦门大学 微流控芯片及其试剂的储存和释放方法
US11311882B2 (en) 2017-09-01 2022-04-26 Miroculus Inc. Digital microfluidics devices and methods of using them
WO2019213583A1 (fr) 2018-05-04 2019-11-07 Abbott Laboratories Procédé d'échantillonnage séquentiel pour améliorer la sensibilité et la cinétique de dosage immunologique d'échantillons de faible volume
WO2020140071A1 (fr) 2018-12-28 2020-07-02 Abbott Laboratories Détection directe de molécules simples sur des microparticules
US11738345B2 (en) 2019-04-08 2023-08-29 Miroculus Inc. Multi-cartridge digital microfluidics apparatuses and methods of use
US11524298B2 (en) 2019-07-25 2022-12-13 Miroculus Inc. Digital microfluidics devices and methods of use thereof
WO2021026402A1 (fr) 2019-08-07 2021-02-11 Abbott Laboratories Composés chimioluminescents pour le multiplexage
WO2021026403A1 (fr) 2019-08-07 2021-02-11 Abbott Laboratories Méthodes de détection de substances interférant avec une analyse et d'augmentation de l'échelle dynamique
CN110918146A (zh) * 2019-12-19 2020-03-27 上海天马微电子有限公司 微流控面板
CN110918145A (zh) * 2019-12-19 2020-03-27 上海天马微电子有限公司 微流控面板及其驱动方法
US20210205813A1 (en) * 2019-12-23 2021-07-08 National Technology & Engineering Solutions Of Sandia, Llc Contactless liquid loading to microfluidic devices
CN111054455A (zh) * 2019-12-26 2020-04-24 上海天马微电子有限公司 微流控芯片及其驱动方法
WO2023122232A1 (fr) * 2021-12-23 2023-06-29 Amgen Inc. Procédés de détection d'une protéine dans un échantillon dans un dispositif fluidique à l'aide d'une spectrométrie de masse
US11772093B2 (en) 2022-01-12 2023-10-03 Miroculus Inc. Methods of mechanical microfluidic manipulation
US11857961B2 (en) 2022-01-12 2024-01-02 Miroculus Inc. Sequencing by synthesis using mechanical compression

Also Published As

Publication number Publication date
WO2007136386A3 (fr) 2008-11-27

Similar Documents

Publication Publication Date Title
WO2007136386A2 (fr) Préparation d'échantillons sur puce à base de gouttelettes destinée à la spectrométrie de masse
US5705813A (en) Integrated planar liquid handling system for maldi-TOF MS
Couvillion et al. New mass spectrometry technologies contributing towards comprehensive and high throughput omics analyses of single cells
US8187864B2 (en) Exchangeable sheets pre-loaded with reagent depots for digital microfluidics
US7303727B1 (en) Microfluidic sample delivery devices, systems, and methods
US20060252047A1 (en) Device and method for analysis of samples using a combined sample treatment and sample carrier device
US20020045270A1 (en) Structured biosample support plates for mass spectroscopic analyses and procedures for manufacturing and use
US9211542B2 (en) High-density sample support plate for automated sample aliquoting
Lee et al. Microfluidics with MALDI analysis for proteomics—A review
CA2563054A1 (fr) Dispositif de presentation d'echantillons
CN110621405B (zh) 用于样本分析的方法和装置
EP2190548A1 (fr) Procédés de séparation de macromolécules fonctionnalisées
Ha et al. Faster, better, and cheaper: harnessing microfluidics and mass spectrometry for biotechnology
US20110095201A1 (en) Electrowetting sample presentation device for matrix-assisted laser desorption/ionization mass spectrometry and related methods
JP2005513454A (ja) Ｆｆｅアレイディスペンサー
US20050029440A1 (en) Machine and method for processing biomolecules
Smithers et al. Interfacing microfluidics with information-rich detection systems for cells, bioparticles, and molecules
US20060263259A1 (en) Apparatus and method for sample preparation and direct spotting eluants onto a MALDI-TOF target
AU2003304421B2 (en) Sample presentation device with differing wettability
Laurell et al. Proteomics–protein profiling technology: the trend towards a microfabricated toolbox concept
Chen et al. Sample preparation for MALDI mass spectrometry using an elastomeric device reversibly sealed on the MALDI target
Laurell et al. Microfluidic components for protein characterization
US20210102954A1 (en) Picoliter droplet sample processing and deposition for mass spectrometry
Wheeler et al. Electrowetting-on-dielectric for analysis of peptides and proteins by matrix assisted laser desorption/ionization mass spectrometry
Yang Microfluidic Interfaces for Mass Spectrometry: Methods and Applications

Legal Events

Date Code Title Description
NENP Non-entry into the national phase in:

Ref country code: DE

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 06851265

Country of ref document: EP

Kind code of ref document: A2

122 Ep: pct application non-entry in european phase

Ref document number: 06851265

Country of ref document: EP

Kind code of ref document: A2