The document discusses various techniques for blood cell separation at the point-of-care, including conventional centrifugation methods and newer microfluidic approaches. Size-based separation techniques discussed include obstacle-based deterministic lateral displacement and successive sieving, while affinity-based techniques include fluorescence labeling, adhesion to antibody-coated surfaces, magnetophoresis using magnetic beads, and dielectrophoresis using non-uniform electric fields. Challenges include achieving sufficient specificity, efficiency, and scale for point-of-care applications.
Molecular weight determination and Characterization of Enzymes Ayushisomvanshi1
This presentation shows about how the determination of molecular weight is done and what are the different ways or method to determine the molecular weight. This presentation also tells about the enzymes and its characterization.
Biosensors are the analytical device that are used to measure the concentration of analye , these type of biosensors are made with conjugation of enzymes as a biological eliment to quantify a (bio)chemical substance / analyte are reffered to as Enzyme-probe Biosensors .
Biosensors are of many types but focusing on Enzyme biosensors there are 4 main types which are briefly described in this power point presentation .
Multienzyme System: A complex of enzymes within a cell that form a reaction sequence of a biochemical pathway so that the product of the first enzyme reaction is transferred directly to the next enzyme and immediately undergoes a second reaction, and so on.
Molecular weight determination and Characterization of Enzymes Ayushisomvanshi1
This presentation shows about how the determination of molecular weight is done and what are the different ways or method to determine the molecular weight. This presentation also tells about the enzymes and its characterization.
Biosensors are the analytical device that are used to measure the concentration of analye , these type of biosensors are made with conjugation of enzymes as a biological eliment to quantify a (bio)chemical substance / analyte are reffered to as Enzyme-probe Biosensors .
Biosensors are of many types but focusing on Enzyme biosensors there are 4 main types which are briefly described in this power point presentation .
Multienzyme System: A complex of enzymes within a cell that form a reaction sequence of a biochemical pathway so that the product of the first enzyme reaction is transferred directly to the next enzyme and immediately undergoes a second reaction, and so on.
Antibody purification – what you need to know to use antibodies effectivelyExpedeon
In this webinar Dr Andy Lane discusses the various methods available for purifying antibodies from different sources, and explains why it is vitally important to understand how your antibodies have been purified to know what you can do with them, either within assays or for further processing such as conjugation to dyes and enzymes.
Two-dimensional gel electrophoresis (2-DE) is considered a powerful tool for proteomics work. 2-DE separates proteins depending on two differ steps: the first one is called isoelectric focusing (IEF) which separates proteins according to isoelectric points (pI); the second step is SDS-polyacrylamide gel electrophoresis (SDS-PAGE) which separates proteins based on the molecular weights.
Our website: www.creative-proteomics.com
2D-Electrophoresis is an important technique that is being used extensively in the Biochemistry and molecular biology for the quantification of different bio-molecules. It is also used in the different researches like cancer study etc. This presentation covers the introduction, sample preparation, main methodology and steps, staining techniques, applications, cost and availability across Pakistan. It also explains that why there is a need to replace the Electrophoresis with 2D electrophoresis. The main purpose of this effort is to highlight the main points about 2D-Electrophoresis.
Introduction, Principle, Instrumentation and Applications of SDS-PAGEMohammed Mubeen
The following presentation contains helpful information regarding SDS-PAGE, including the history, introduction, principle, instrumentation, advantages and applications of SDS-PAGE.
Antibody purification – what you need to know to use antibodies effectivelyExpedeon
In this webinar Dr Andy Lane discusses the various methods available for purifying antibodies from different sources, and explains why it is vitally important to understand how your antibodies have been purified to know what you can do with them, either within assays or for further processing such as conjugation to dyes and enzymes.
Two-dimensional gel electrophoresis (2-DE) is considered a powerful tool for proteomics work. 2-DE separates proteins depending on two differ steps: the first one is called isoelectric focusing (IEF) which separates proteins according to isoelectric points (pI); the second step is SDS-polyacrylamide gel electrophoresis (SDS-PAGE) which separates proteins based on the molecular weights.
Our website: www.creative-proteomics.com
2D-Electrophoresis is an important technique that is being used extensively in the Biochemistry and molecular biology for the quantification of different bio-molecules. It is also used in the different researches like cancer study etc. This presentation covers the introduction, sample preparation, main methodology and steps, staining techniques, applications, cost and availability across Pakistan. It also explains that why there is a need to replace the Electrophoresis with 2D electrophoresis. The main purpose of this effort is to highlight the main points about 2D-Electrophoresis.
Introduction, Principle, Instrumentation and Applications of SDS-PAGEMohammed Mubeen
The following presentation contains helpful information regarding SDS-PAGE, including the history, introduction, principle, instrumentation, advantages and applications of SDS-PAGE.
[Webinar] Re-Strategezing for a Successful ICD-10 / 2015 TransitionPhoenix Health Systems
In this 45-minute webinar, You will hear from top members of our ICD-10 team, Thomas Grove, Principal and D’Arcy Guerin Gue, Executive Vice President, as they discuss how to ensure the smoothest possible conversion, by restructuring your strategy in eleven critical areas.
Find the recording here - http://landing.phoenixhealth.com/icd-10-2015-webinar
OTN tour 2015 Oracle Enterprise Manager 12c – Proof of ConceptAndrejs Vorobjovs
Why we are talking about this
How – minimal survival kit
Database provisioning:
Database provisioning
Pluggable database provisioning
Schema provisioning
Middleware provisioning:
New instance installation
Instance cloning
Integration provisioning
Restrictions
Conclusion
Q&A
Peteris Arajs
Technology Architecture Associate Manager at Accenture
More than 15 years experience in IT industry with main focus to:
- DB design, analysis, development and performance tuning
- Oracle eBusiness Suite
- Oracle Middleware
Also experienced in all stages of software development life cycle (SDLC) from business requirements and technical definitions to development, testing and production support.
Alex Nemirovskis
Technology Architecture Associate Manager at Accenture
More than 19 years experience in IT industry with main focus to:
- DB design, analysis, development and performance tuning
- DWH / ETL / BI / Analytics
- Oracle ADF
Also experienced in all stages of software development life cycle (SDLC) from business requirements and technical definitions to development, testing and production support.
Proof of Concept with Real Application Testing 12cLuis Marques
Evaluate how certain real world database workload behaves on different I/O subsystem, processors and
architecture or the coexistence with other databases is the goal of a Proof of Concept. The need of testing
real production workloads to eliminate uncertainty with help of techniques like Workload Folding, Time
Shifting and Schema Remapping, this talk will produce evidence that exploring Real Application Testing
features in 12c leverage what can be accomplished by a Proof of Concept.
VMWare Lab For Training and Testing: As our IT infrastructure grows in complexity and new products are released every so often, we are faced with the same expectations to deliver tested and innovative IT environment while reducing the cost and speeding up the whole process of testing and training. Now, this is a real challenge if you don’t have all of the resources!
So, how can you reduce the cost, the time to plan, install, configure, validate and support complex virtual labs for training, practice or proof of concept? It’s time for a breakthrough, innovative solution…called: Cloud-Based VMWare Lab http://viadmin.com/pages/VMWare-Practice-Lab-at-Home.html!
BLOOD FILM EXAMINATION: ITS RECENT INVESTIGATIVE METHODOLOGY IN THE DIAGNOSIS...Chibueze Nwudele
In patient’s care, diagnostic formulations rest on a tripod consisting of clinical history, physical examination and laboratory investigation.
The literature reveals that as much as 70% of clinical decisions and diagnosis are supported by laboratory medicine (WHO, 2015; Adewoyin & Nwogoh, 2014).
Peripheral blood film (PBF) is a basic and a highly informative haematological tool at the clinician’s disposal in screening, diagnosis and monitoring of disease progression and therapeutic response.
An adept understanding of peripheral blood interpretation is important for a successful clinical practice (Adewoyin & Nwogoh, 2014).
Therefore, the ability to prepare, stain and report correct findings of a peripheral blood film is a skill that every Medical Laboratory Scientist should desire and study extensively to get expertise in.
This is important as its role in the investigations and diagnosis of diseases mostly anaemia and most other haematological disorders cannot be over emphasized. To me, it is the hallmark of haematology.
Describes the mission of laboratory services, the phases of performing laboratory tests, factors affecting laboratory tests and strorage and transport of laboratory samples
Internship - FMRI, Gurgaon (Dec '18 - Jan '19) AryanDugar
This is a presentation detailing the knowledge I acquired at my observational internship in the Clinical Pathology laboratory at Fortis Memorial Research Institute, Gurgaon, India.
Resealed erythrocytes as a novel delivery carrierMariamZewail
Resealed erythrocytes are effective and safe drug carriers for targeted and sustained drug delivery. Drugs can be easily entrapped into erythrocytes by several techniques. Resealed erythrocytes can be used a carrier for drugs, enzyme replacement therapy etc. However, the concept needs further optimization to be converted into a regular drug delivery system.
Resealed erythrocytes as a novel delivery carrierMariamZewail
Resealed erythrocytes can be obtained due to the different responses of red blood cells in response to different pHs. Resealed erythrocytes have several applications as a drug or enzyme carrier.
2. Point of care (POC) testing 11/24/2008 ■ Does not require permanent, dedicated space. ■ Eliminates need for nursing administrative efforts. ■ Focus on direct needs of patients. ■ Performed in proximity to the patient. ■ Provides “Fast Facts” to quickly identify and contain a disease. ■ Transfer from Lab/ field testing environment. ■ Reduce Cost of Diagnostic and testing ■ Efficient Data Management & Patients record keeping. ■ Flexibility of application (can be configured for other applications) ■ Plug-and-play interoperability for medical device communications (IEEE Std) ■ Weight, Size, Form, Battery Life, Reliability, Accuracy, Multiple input ■ Bio-Degradable/Disposable Sample or Applicator. AZ (Source: Building blocks for point-of-care boom, Ian Macfarlane & Fred Davis)
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5. 11/24/2008 Blood Composition & Diversity Cell population and subpopulation in normal blood 7- 20μm 6.5 to 8 4μm 1- 4μm 12- 15μm 10- 15μm 10- 15μm 10- 15μm AZ . Cell diameters too close to each other. POC resolution must be better less than 1 um (Blood-On-A-Chip, “Annual Rev. Mehmet Toner & Daniel Irima Biomed. Eng. 2005 7:77-103 ) Cell population diversity adds to the difficulty of separation
6. 11/24/2008 Blood Composition & Detection AZ RBC: Carry O2 WBC: Do not carry O2 (Wheater’s Functional Histology) Blood cells are different by Size, Composition, Color, Weight, Function, and percentage in sample RBC WBCs NEUTROPHILIC EOSINOPHILIC BASOPHILIC LYMPHOCYTE MONOCYTE MONOCYTE (Davidson , 2007) Cell Detection is expensive and Lab-oriented (Wheater’s Functional Histology)
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12. 11/24/2008 Multi-Functionality of POC AZ ( “ Understanding Complete Blood Count “ (CBC) Report; Sonora Quest Laboratories, Arizona, CA.) Many Disorders One Platform GOAL: Maximize associated disorders through Similar Processes of separation and detection
13. Cell Separation Techniques Research & Development 11/24/2008 ■ Cell separation techniques can be broadly classified into two categories: B. Sized-Based Methods: - Relatively fast and simple. - Type of cells is determined and separated according to their cell size, shape and other physical properties - Disadvantage of this method is its low specificity for cell separation, - Separation is not too sensitive to small size variation of cells. A. Affinity-Based Methods: - Particle separation due to affinity of antigen to antibodies in the sample with high specificity and selectivity - The isolated cells may suffer from damages. - High cost and complicated processes such as immunoreactions and elution (extraction by means of solvents) of cells from the capturing antibodies create challenges (Zheng, Siyang, Raylene Yung, et al. 2005) AZ (Zheng, Siyang, Raylene Yung, et al. 2005)
14. Cell Separation Techniques Research & Development 11/24/2008 ■ Cell separation techniques can be broadly classified into two categories: A. Techniques based on size, shape & density. B. Techniques based on cell affinity (chemical, electrical, or magnetic) Magnetophoresis Electrophoresis Adhesion-Based Florescence-Based Capillary Electrophoresis & Capillary cIEF Electro-hydrodynamic AZ Obstacle & Sieve (Zheng, Siyang, Raylene Yung, et al. 2005)
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22. 11/24/2008 Capillary electrophoresis (CE) AZ ■ Uses capillaries (e.g. porous fused-silica) to separate a complex array of large/small molecules ■ High electric field strengths are used to separate molecules based on differences in charge, size and hydrophobicity. ■ Apply pressure, vacuum or voltage to end of the capillary vial sample too separate. ■ Types of capillary and electrolytes determine the separation techniques: Surfactants are added to the buffer solution at concentrations that form micelles. Separation Principle: differential partition between the micelle and the solvent. Micelles: An aggregate of surfactant molecules dispersed in a liquid colloid. ○ Micellar Electrokinetic Capillary Chromatography (MECC ) Solutes partition with moving oil droplets in buffer. ○ Micro Emulsion Electrokinetic Chromatography (MEEKC) Include electro-osmosis, electrophoresis and chromatography (adsorptive materials separation) ○ Electro-kinetic Chromatography (EKC): Based on the migration of the sample components between leading and terminating electrolytes ○ Isotacho-phoresis (ITP) Electrophoresis in a pH gradient generated between the cathode and anode. A solute will migrate to a point where its net charge is zero. Sample focused into tight zone. ○ Capillary Isoelectric Focusing (CIEF) Uses polymers in solution to create a molecular sieve. ○ Capillary Gel Electrophoresis (CGE) Based on differences in the charge-to-mass ratio of the analytes ○ Capillary Zone Electrophoresis (CZE ( Beckman Coulter Corporation, “Capillary Electrophoresis: A Simple Technique”)
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24. 11/24/2008 Summary of Methods ● PH dependent ● Low efficiency ● Requires Pressure or Vacuum. ► Laminar Flow ► Moderate throughput Capillary Electrophoresis : Capillary Isoelectric Focusing (cIEF) ● Packed bed design; Difficult to miniaturized ● Antibody tags : Requires lab preparation ● Cell populations have same size or density ► No need for incubation of staring cell ► Provides high purity (>95%) ► Provide high throughput (108-109 cells/hour) Surface Adhesion ► Electro-osmotic flow potential ► Laminar Flow Florescence Activated Cell Sorting (FACS) ► Post Size and Diameter ► Particle size differentiation ► Used mostly on protein & Peptides Electrophoresis ● Require Cell-Specific Marker. ● Requires exact cell position relative to the magnetic field, ● Low efficiency in general. ● Maximum two particle sorting ● AC Power ● Antibody tags : Requires lab preparation ● High Cost (Not feasible commercially) ● Difficult to miniaturize ● Non-Intrusive method: Blood must be deoxygenated. ► Particle size differentiation ► Magnetic properties ► High efficiency possible Magnetophoresis. - Intrusive - Non-Intrusive ● Require Cell-Specific Marker. ● Isolated cells suffer from damage. ● Low efficiency ● High Cost (Not feasible commercially) ► Laminar Flow ► Continuous (flow through) system Magnetic Activated Cell Sorting (MACS) ● Requires high electric Field ● Difficult to miniaturize ► High efficiency Electro-dynamic Flow ● Low specificity of cell separation. ● Insensitive to small differences in Size & Form ► Laminar Flow ► Cell-Specific Markers not required Size-Based WEAKNESS A Good POC Candidate? IMPORTANT PARAMETERS & FEATURES SEPARATION METHOD` AZ
25. Challenges in blood cells separation and detection 11/24/2008 ■ Challenge: Massive numbers & diversity of cell types complicate precise identification of the target cells from blood sample. Approach: Multiple sampling (within the same package) and using similar method to identify various cell properties. ■ Challenge: Many cells can only be identified by the presence of specific protein markers (Antigen/Antibody) on the surface of cells Approach: Use intrinsic cell properties (Mobility, polarity etc.) that eliminates need for specific markers. Integrate a combination of cell separation techniques in single platform. ■ Challenge: Some processes require longer time of sorting & detection (e.g. sample culture) Approach: New innovations are key to success in reducing time of processes ■ Challenge: Minimum handling to reduce contamination and exposure Approach: Self contained, discard able, bio-degradable test sample; Lab-on-Chip ■ Challenge: Test Package is inadequate for Field use ( Too big, AC power, handling, Non-portable) Approach: Reduce complexity of design with fresh scientific approach. DC Power. Portable platform ■ Challenge: High Cost($) & Low efficiency Approach: Simple Design & Instrumentation (Mfg. for COTS). Improved detection system ■ Challenge: High Reliability & Repeatability ( particle count > 50000 Cells/sec) Approach: Miniaturization of improved instruments. New techniques of detection. Increase precision in nanoscale domain (CNT, naowires,..etc) Joseph/AZ