Cell culture refers to the removal of cells from an animal or plant and their subsequent growth in a favorable artificial environment. The cells may be removed from the tissue directly and disaggregated by enzymatic or mechanical means before cultivation, or they may be derived from a cell line or cell strain that has already been established.
More cell culture techniques and best practices here. http://owl.li/dgS2Y
Cell culture involves growing cells under controlled conditions. Key developments include the use of antibiotics to prevent contamination, trypsin to detach adherent cells for subculturing, and defined culture media. Cells are typically maintained through serial passaging when confluent. Cells can be cryopreserved for long-term storage in liquid nitrogen. Common cell lines include HeLa, HEK293, MCF-7, and Vero cells. Contaminants include mycoplasma, bacteria, and cross-contamination between cell lines. Basic equipment includes laminar flow hoods, incubators, refrigerators, and microscopes.
Cell culture is the process of growing cells under controlled conditions outside of their natural environment. Key developments in cell culture technology include the use of antibiotics to prevent contamination, trypsin to detach adherent cells for subculturing, and chemically defined culture media. Cell culture is used in a variety of areas including basic research, toxicity testing, cancer research, virology, genetic engineering, and gene therapy. Successful cryopreservation of cell lines involves slow freezing and quick thawing to minimize ice crystal formation and damage to cells.
Principles & Applications of cell viability assays (MTT Assays)VidyaNani
This document discusses cell viability assays, specifically focusing on principles and applications of MTT assays. It defines cell viability and describes various types of cell viability assays including dye exclusion, colorimetric, fluorometric, luminometric, and flow cytometric assays. The document provides details on the MTT assay, including its principle, protocol, and applications for measuring cell proliferation, cytotoxicity, and metabolic activity. The MTT assay is a common colorimetric assay that measures the reduction of yellow MTT by mitochondrial dehydrogenases in viable cells to produce purple formazan crystals.
Cell lines are permanently established cell cultures that can proliferate indefinitely. They are derived from primary cell cultures which have a finite lifespan. Continuous cell lines have undergone transformation to proliferate indefinitely, unlike finite primary cultures. Common types of cell lines include normal and transformed cell lines of epithelial, fibroblast, or lymphoblast morphology derived from human or animal tissues. Primary cultures are used to establish cell lines which are then used for applications like drug screening, toxicity testing, and cancer research.
The document provides information on principles of cell culture. It discusses the history of cell culture, beginning with Roux maintaining embryonic chick cells in 1885. It describes the typical equipment used in cell culture like laminar flow cabinets and incubators. The document outlines the different types of cell cultures like primary cultures derived directly from tissue and continuous cultures that can be serially passaged. It explains concepts like passaging cells to maintain and expand cultures. The document also discusses cryopreservation to store cells long-term in liquid nitrogen. Common cell lines used in research are also mentioned like HeLa and MCF-7 cells. Contamination in cell culture can occur from bacteria, fungi or mycoplasma and affect cell growth. Strict aseptic
Cell culture involves growing cells under controlled conditions. Key developments include the use of antibiotics to prevent contamination, trypsin to detach adherent cells for subculturing, and defined culture media. Cells are typically maintained through serial passaging when confluent. Cells can be cryopreserved for long-term storage in liquid nitrogen. Common cell lines include HeLa, HEK293, MCF-7, and Vero cells. Contaminants include mycoplasma, bacteria, and cross-contamination between cell lines. Basic equipment includes laminar flow hoods, incubators, refrigerators, and microscopes.
Cell culture is the process of growing cells under controlled conditions outside of their natural environment. Key developments in cell culture technology include the use of antibiotics to prevent contamination, trypsin to detach adherent cells for subculturing, and chemically defined culture media. Cell culture is used in a variety of areas including basic research, toxicity testing, cancer research, virology, genetic engineering, and gene therapy. Successful cryopreservation of cell lines involves slow freezing and quick thawing to minimize ice crystal formation and damage to cells.
Principles & Applications of cell viability assays (MTT Assays)VidyaNani
This document discusses cell viability assays, specifically focusing on principles and applications of MTT assays. It defines cell viability and describes various types of cell viability assays including dye exclusion, colorimetric, fluorometric, luminometric, and flow cytometric assays. The document provides details on the MTT assay, including its principle, protocol, and applications for measuring cell proliferation, cytotoxicity, and metabolic activity. The MTT assay is a common colorimetric assay that measures the reduction of yellow MTT by mitochondrial dehydrogenases in viable cells to produce purple formazan crystals.
Cell lines are permanently established cell cultures that can proliferate indefinitely. They are derived from primary cell cultures which have a finite lifespan. Continuous cell lines have undergone transformation to proliferate indefinitely, unlike finite primary cultures. Common types of cell lines include normal and transformed cell lines of epithelial, fibroblast, or lymphoblast morphology derived from human or animal tissues. Primary cultures are used to establish cell lines which are then used for applications like drug screening, toxicity testing, and cancer research.
The document provides information on principles of cell culture. It discusses the history of cell culture, beginning with Roux maintaining embryonic chick cells in 1885. It describes the typical equipment used in cell culture like laminar flow cabinets and incubators. The document outlines the different types of cell cultures like primary cultures derived directly from tissue and continuous cultures that can be serially passaged. It explains concepts like passaging cells to maintain and expand cultures. The document also discusses cryopreservation to store cells long-term in liquid nitrogen. Common cell lines used in research are also mentioned like HeLa and MCF-7 cells. Contamination in cell culture can occur from bacteria, fungi or mycoplasma and affect cell growth. Strict aseptic
This document discusses methods for evaluating the cytotoxicity of nanoparticles. It describes several common cytotoxicity assays including MTT, WST, trypan blue exclusion, and assays using dehydrogenases. The MTT assay measures mitochondrial activity and is widely used. WST assays use water-soluble reagents and do not require crystal solubilization. Dehydrogenase assays offer high sensitivity by measuring multiple cell elements. The document also provides examples of studies that used these assays to evaluate the cytotoxicity of silver nanoparticles, magnetic nanoparticles, and other nanomaterials.
Cell lines need to be routinely maintained and stored long-term to preserve their valuable characteristics. Routine maintenance involves periodic medium changes and subculturing depending on the growth rate of the specific cell line. Long-term storage is achieved through cryopreservation, where cells are frozen at temperatures below -100°C. This process aims to minimize ice crystal formation and associated cell damage through slow freezing in the presence of cryoprotectants like DMSO or glycerol. Proper freezing and storage methods help preserve cell lines for future use and distribution.
This document provides an overview of flow cytometry and fluorescence-activated cell sorting (FACS). It describes flow cytometry as a technique for measuring physical and chemical characteristics of cells as they flow in a fluid stream, allowing for single cell analysis. FACS extends this by using fluorescence to identify cell characteristics and sort cells into separate collections based on these characteristics. The key components of a flow cytometer are described as lasers, optics including filters and detectors, fluidics to hydrodynamically focus cells, and electronics to convert optical signals to digital data. Applications including cell phenotyping, apoptosis analysis, and cell cycle analysis are discussed. Cell sorting and quantitative analysis of cell cycle phases are also summarized.
The document discusses cell lines, including their origins from primary cell cultures. A cell line is permanently established and will proliferate indefinitely, unlike a cell strain which has finite divisions. Cell lines can be finite or continuous. Continuous cell lines are immortalized through spontaneous or induced genetic changes. Common sources of cell lines include ATCC, ECACC, and NCCS. Selection of cell lines considers species, growth characteristics, stability, and availability. Cell lines must be maintained under controlled conditions like temperature and pH. Applications include drug screening and cancer research. Commonly used cell lines include HEK293 derived from embryonic kidney cells.
This document describes the MTT assay, a colorimetric assay used to measure cell viability and cytotoxicity. The MTT assay works by using the enzyme mitochondrial dehydrogenase in living cells to reduce the yellow tetrazolium dye MTT to purple insoluble formazan. The amount of formazan produced is directly proportional to the number of viable cells. The document outlines the principle, reagents, procedure, troubleshooting, advantages, and disadvantages of the MTT assay. Commonly available MTT assay kits are also listed.
Cell viability and proliferation assays measure aspects of cellular health and function, such as membrane integrity, metabolic activity, and DNA synthesis. Common assays include MTT, which measures mitochondrial activity; ATP assays, which measure ATP concentration as a marker of viability; Sulforhodamine B, which binds cellular proteins to measure biomass; and propidium iodide staining, which detects compromised membranes. These assays are useful for screening drug toxicity and effects on cell growth.
Cell synchronization helps in obtaining distinct sub population of cells representing different stages of cell cycle.It helps in collecting population wide data of cells progressing through various stages of cell cycle. Immortalization, refers to cells having capability of undergoing cell division infinitely. Immortal cells are particularly preferred in cell culture to enable long time storage and use. This presentation teaches about cell synchronization, methods of cell synchronization, cellular transformation, immortalization and mechanism of immortalization.
This document discusses different types of cell culture, including primary culture, secondary culture, cell lines, and established cell line culture. It describes the process of isolating and culturing cells from tissue samples, including enzymatic and mechanical disaggregation of tissues. The importance of physical environment and culture media for growing cells in vitro is highlighted. Different types of culture media like serum-containing, serum-free, and chemically defined media are also summarized.
Secondary cell cultures refer to cells that have been subcultured, or transferred, from a primary culture to a new culture vessel. Subculturing provides fresh nutrients and space for continuously growing cell lines. Cell lines can be finite or continuous depending on their lifespan in culture. Characterization of cell lines is important to confirm identity and purity through analysis of biochemical, genetic, and chromosomal parameters such as karyotyping.
chimeric antigen receptor, its structure and role in killing tumor cells,mechanism of antitumor killing, car's in clinic,evolution of cars and new chimeric antigen models
This document discusses cell culture in animal biotechnology. It provides an overview of the history of cell culture from the late 1800s to modern applications. Some key points covered include:
1) Cell culture refers to growing cells in a controlled artificial environment outside of their natural environment. Examples of cell types grown include fibroblasts, lymphocytes, and cells from various tissues.
2) Important milestones in the history of cell culture include Roux maintaining embryonic chick cells in 1885 and Carrel introducing strict aseptic techniques allowing long-term cell culture in 1913.
3) Cell culture has various applications including use as model systems, in toxicity testing, cancer research, virology, drug development, and gene therapy.
FACS and MACS with their applications in biological research.Deepak Agarwal
Flow cytometry (FACS) and magnetic activated cell sorting (MACS) are techniques used to analyze and separate cells based on their physical and chemical characteristics. FACS uses lasers to detect cell properties and sort cells into containers one by one, while MACS uses magnetic microbeads attached to cells to separate them in high gradient magnetic fields. These techniques have various applications in research including identifying stem cells, characterizing cancer cells, studying cell cycles, and isolating cell populations for further analysis.
Bioreactors for animal cell suspension cultureGrace Felciya
This document discusses bioreactors for animal cell suspension culture. It begins by introducing animal cell culture and some key developments that enabled it. There are two main types of culture: primary culture using explants or enzymes, and secondary culture which is derived from primary culture. Cells can be anchorage-dependent, growing in monolayers, or non-anchorage dependent, growing in suspension. Bioreactors provide conditions for mass cultivation of suspension cells. Properties of animal cells require gentle mixing and aeration in bioreactors. Common bioreactor types for suspension culture include stirred tank, continuous flow, and airlift fermentors. Perfusion culture allows continuous medium exchange to achieve high cell densities and productivity.
PRINCIPLES AND APPLICATIONS OF CELL VIABILITY ASSAY (MTT ASSAY)Durgadevi Ganesan
The document discusses the principles and applications of MTT cell viability assays. The MTT assay is a colorimetric assay that measures the reduction of MTT by mitochondrial succinate dehydrogenase in metabolically active cells to form an insoluble purple formazan product. The amount of formazan produced is directly proportional to the number of viable cells and can be used to measure cell proliferation, viability, and cytotoxicity in response to drugs or other factors. The MTT assay is inexpensive, rapid, quantitative, and reproducible, making it well-suited for applications like cytotoxicity testing, drug screening, and measuring responses to growth factors.
8. Biology and characterization of cultured cellsShailendra shera
Immediate environment and environment of surrounding medium governs the various properties of cell. The in vitro condition markedly affects the cellular property of cultured cells. For e.g. Reduction in Cell–cell and cell-material interaction. Therefore, it is imperative to develop understanding of biology of cells in response to various environmental conditions. Characterization of cells helps to identify the origin, purity and authenticity of cells and cell lines.
This document provides information about cell lines. It defines different types of cell lines including primary cell cultures, continuous cell lines, and transient cell lines. It discusses the HeLa cell line which was the first human cancer cell line. It outlines the process for generating stable cell lines including transfection, selection, cloning, and screening for stability. Steps for freezing and storing cell lines are described. Considerations for choosing an appropriate cell line for experiments are presented. Details are given about specific commonly used cell lines like 3T3, HEK293, PC12, CHO, and BC1. References for further information on cell line generation and characterization are provided.
Tissue culture involves growing cells, tissues, or organs outside of their natural biological context. There are three main types of tissue culture: cell culture, explant culture, and organ culture. Cell culture involves growing isolated cells, either in an adherent monolayer or in suspension. Eplant culture grows fragments of tissue on a substrate, while organ culture maintains the three-dimensional structure of whole organs. Tissue culture has advantages like control over the environment and scale-up potential, but cells may lose differentiated characteristics over time.
This document discusses methods for evaluating the cytotoxicity of nanoparticles. It describes several common cytotoxicity assays including MTT, WST, trypan blue exclusion, and assays using dehydrogenases. The MTT assay measures mitochondrial activity and is widely used. WST assays use water-soluble reagents and do not require crystal solubilization. Dehydrogenase assays offer high sensitivity by measuring multiple cell elements. The document also provides examples of studies that used these assays to evaluate the cytotoxicity of silver nanoparticles, magnetic nanoparticles, and other nanomaterials.
Cell lines need to be routinely maintained and stored long-term to preserve their valuable characteristics. Routine maintenance involves periodic medium changes and subculturing depending on the growth rate of the specific cell line. Long-term storage is achieved through cryopreservation, where cells are frozen at temperatures below -100°C. This process aims to minimize ice crystal formation and associated cell damage through slow freezing in the presence of cryoprotectants like DMSO or glycerol. Proper freezing and storage methods help preserve cell lines for future use and distribution.
This document provides an overview of flow cytometry and fluorescence-activated cell sorting (FACS). It describes flow cytometry as a technique for measuring physical and chemical characteristics of cells as they flow in a fluid stream, allowing for single cell analysis. FACS extends this by using fluorescence to identify cell characteristics and sort cells into separate collections based on these characteristics. The key components of a flow cytometer are described as lasers, optics including filters and detectors, fluidics to hydrodynamically focus cells, and electronics to convert optical signals to digital data. Applications including cell phenotyping, apoptosis analysis, and cell cycle analysis are discussed. Cell sorting and quantitative analysis of cell cycle phases are also summarized.
The document discusses cell lines, including their origins from primary cell cultures. A cell line is permanently established and will proliferate indefinitely, unlike a cell strain which has finite divisions. Cell lines can be finite or continuous. Continuous cell lines are immortalized through spontaneous or induced genetic changes. Common sources of cell lines include ATCC, ECACC, and NCCS. Selection of cell lines considers species, growth characteristics, stability, and availability. Cell lines must be maintained under controlled conditions like temperature and pH. Applications include drug screening and cancer research. Commonly used cell lines include HEK293 derived from embryonic kidney cells.
This document describes the MTT assay, a colorimetric assay used to measure cell viability and cytotoxicity. The MTT assay works by using the enzyme mitochondrial dehydrogenase in living cells to reduce the yellow tetrazolium dye MTT to purple insoluble formazan. The amount of formazan produced is directly proportional to the number of viable cells. The document outlines the principle, reagents, procedure, troubleshooting, advantages, and disadvantages of the MTT assay. Commonly available MTT assay kits are also listed.
Cell viability and proliferation assays measure aspects of cellular health and function, such as membrane integrity, metabolic activity, and DNA synthesis. Common assays include MTT, which measures mitochondrial activity; ATP assays, which measure ATP concentration as a marker of viability; Sulforhodamine B, which binds cellular proteins to measure biomass; and propidium iodide staining, which detects compromised membranes. These assays are useful for screening drug toxicity and effects on cell growth.
Cell synchronization helps in obtaining distinct sub population of cells representing different stages of cell cycle.It helps in collecting population wide data of cells progressing through various stages of cell cycle. Immortalization, refers to cells having capability of undergoing cell division infinitely. Immortal cells are particularly preferred in cell culture to enable long time storage and use. This presentation teaches about cell synchronization, methods of cell synchronization, cellular transformation, immortalization and mechanism of immortalization.
This document discusses different types of cell culture, including primary culture, secondary culture, cell lines, and established cell line culture. It describes the process of isolating and culturing cells from tissue samples, including enzymatic and mechanical disaggregation of tissues. The importance of physical environment and culture media for growing cells in vitro is highlighted. Different types of culture media like serum-containing, serum-free, and chemically defined media are also summarized.
Secondary cell cultures refer to cells that have been subcultured, or transferred, from a primary culture to a new culture vessel. Subculturing provides fresh nutrients and space for continuously growing cell lines. Cell lines can be finite or continuous depending on their lifespan in culture. Characterization of cell lines is important to confirm identity and purity through analysis of biochemical, genetic, and chromosomal parameters such as karyotyping.
chimeric antigen receptor, its structure and role in killing tumor cells,mechanism of antitumor killing, car's in clinic,evolution of cars and new chimeric antigen models
This document discusses cell culture in animal biotechnology. It provides an overview of the history of cell culture from the late 1800s to modern applications. Some key points covered include:
1) Cell culture refers to growing cells in a controlled artificial environment outside of their natural environment. Examples of cell types grown include fibroblasts, lymphocytes, and cells from various tissues.
2) Important milestones in the history of cell culture include Roux maintaining embryonic chick cells in 1885 and Carrel introducing strict aseptic techniques allowing long-term cell culture in 1913.
3) Cell culture has various applications including use as model systems, in toxicity testing, cancer research, virology, drug development, and gene therapy.
FACS and MACS with their applications in biological research.Deepak Agarwal
Flow cytometry (FACS) and magnetic activated cell sorting (MACS) are techniques used to analyze and separate cells based on their physical and chemical characteristics. FACS uses lasers to detect cell properties and sort cells into containers one by one, while MACS uses magnetic microbeads attached to cells to separate them in high gradient magnetic fields. These techniques have various applications in research including identifying stem cells, characterizing cancer cells, studying cell cycles, and isolating cell populations for further analysis.
Bioreactors for animal cell suspension cultureGrace Felciya
This document discusses bioreactors for animal cell suspension culture. It begins by introducing animal cell culture and some key developments that enabled it. There are two main types of culture: primary culture using explants or enzymes, and secondary culture which is derived from primary culture. Cells can be anchorage-dependent, growing in monolayers, or non-anchorage dependent, growing in suspension. Bioreactors provide conditions for mass cultivation of suspension cells. Properties of animal cells require gentle mixing and aeration in bioreactors. Common bioreactor types for suspension culture include stirred tank, continuous flow, and airlift fermentors. Perfusion culture allows continuous medium exchange to achieve high cell densities and productivity.
PRINCIPLES AND APPLICATIONS OF CELL VIABILITY ASSAY (MTT ASSAY)Durgadevi Ganesan
The document discusses the principles and applications of MTT cell viability assays. The MTT assay is a colorimetric assay that measures the reduction of MTT by mitochondrial succinate dehydrogenase in metabolically active cells to form an insoluble purple formazan product. The amount of formazan produced is directly proportional to the number of viable cells and can be used to measure cell proliferation, viability, and cytotoxicity in response to drugs or other factors. The MTT assay is inexpensive, rapid, quantitative, and reproducible, making it well-suited for applications like cytotoxicity testing, drug screening, and measuring responses to growth factors.
8. Biology and characterization of cultured cellsShailendra shera
Immediate environment and environment of surrounding medium governs the various properties of cell. The in vitro condition markedly affects the cellular property of cultured cells. For e.g. Reduction in Cell–cell and cell-material interaction. Therefore, it is imperative to develop understanding of biology of cells in response to various environmental conditions. Characterization of cells helps to identify the origin, purity and authenticity of cells and cell lines.
This document provides information about cell lines. It defines different types of cell lines including primary cell cultures, continuous cell lines, and transient cell lines. It discusses the HeLa cell line which was the first human cancer cell line. It outlines the process for generating stable cell lines including transfection, selection, cloning, and screening for stability. Steps for freezing and storing cell lines are described. Considerations for choosing an appropriate cell line for experiments are presented. Details are given about specific commonly used cell lines like 3T3, HEK293, PC12, CHO, and BC1. References for further information on cell line generation and characterization are provided.
Tissue culture involves growing cells, tissues, or organs outside of their natural biological context. There are three main types of tissue culture: cell culture, explant culture, and organ culture. Cell culture involves growing isolated cells, either in an adherent monolayer or in suspension. Eplant culture grows fragments of tissue on a substrate, while organ culture maintains the three-dimensional structure of whole organs. Tissue culture has advantages like control over the environment and scale-up potential, but cells may lose differentiated characteristics over time.
The document summarizes a presentation given by Eric Kurzhals on the development of a cutting-edge CHO cell culture medium called the ActiCHO Media System. It discusses typical issues with existing cell culture media, such as being unbalanced and complex. The presentation then outlines how the ActiCHO system addresses these issues by having components designed to work together optimally and being fully chemically defined. It also describes how the system was developed through extensive experimentation and can be easily scaled from shake flasks to 1000L bioreactors with high productivity and batch-to-batch conformity.
The document discusses the development and evaluation of novel chemically-defined media for CHO cell applications. It presents data comparing the performance of Cellvento CHO-200 media to two competitor products. Cell growth and IgG titer were consistently higher in Cellvento CHO-200 over 14 days of fed batch culture. Lot-to-lot consistency was also demonstrated for cell growth and IgG production across three lots each of Cellvento CHO-200 media and feed in batch and fed batch cultures. Near infrared spectroscopy was discussed as a method for media performance prediction. The impacts of powder handling and production processes like milling were evaluated.
Cell Line Development: Reducing timelines and increasing titres fujifilmdiosynth
Cell line development: Reducing timelines and increasing titres by identification of host cell lines with improved characteristics. To develop a mammalian expression platform which rapidly leads to efficient, robust and high quality biomanufacturing processes
This Cell Culture technical tips slideshow includes an overview of lab practices, equipment checklists and techniques to optimise your cell culture experiments.
- Cell culture techniques have developed significantly since 1885 when embryonic chick cells were first maintained alive in saline solution for short periods. Key developments include producing continuous cell lines in 1943 and defining specific growth factor requirements in the 1970s.
- Cell culture is now a widely accepted research tool with applications like studying cell behavior without animal variations, artificial organ development, vaccine development, and drug testing while avoiding ethical issues of animal experimentation.
- Maintaining healthy, reproducible cells requires standardized techniques and controlling contamination remains important.
This document discusses stem cell culture and provides definitions, classifications, and methods for culturing different types of stem cells. It summarizes the history of stem cell research from 1981 to present. It describes embryonic stem cells, adult stem cells including bone marrow and umbilical cord stem cells. Methods are outlined for isolating and culturing stem cells from bone marrow and umbilical cord. Advantages and disadvantages of different stem cell sources are compared.
Cell culture involves growing cells from tissue or organ samples in artificial environments outside of the original organism. There are several stages of cell culture, beginning with isolating tissues through enzymatic or mechanical means. Primary cell cultures have a limited lifespan, while continuous cell lines can proliferate indefinitely. Proper culture conditions require appropriate media, substrates, gases, and temperature/humidity control. Cells may be grown as adherent monolayers or in suspension. Cell culture has many applications including drug development, cancer research, and production of therapeutic products.
The document discusses different types of cell culture used in bioreactors. It describes organ culture, tissue culture, and cell culture. Cell culture involves dispersing tissue enzymatically into a cell suspension that can be grown as a monolayer or in suspension. Continuous cell lines can be propagated indefinitely and have gained immortality through transformation. Bioreactors must provide a well-controlled environment for cell culture and can operate in batch, fed-batch or perfusion modes. Common bioreactor designs include stirred tank, airlift and wave bioreactors.
Cell culture involves removing cells from an animal or plant and growing them in an artificial environment that provides nutrients for growth. Key developments included the use of antibiotics to reduce contamination, enzymes like trypsin to detach adherent cells, and chemically defined media. Cell culture is used for modeling biology, toxicity testing, cancer research, virology, genetic engineering, and producing therapeutic proteins. Proper conditions like temperature, substrate, and media composition are required to keep cells "happy" and growing. Aseptic technique and containment are important to prevent contamination.
1. Researchers transformed green algae cells that grow through photosynthesis (autotrophically) into yellow cells that grow by consuming organic carbon (heterotrophically). This resulted in the altered cells accumulating much higher levels of lipids.
2. The lipid-rich heterotrophic cells were then used to produce biofuels through two methods - fast pyrolysis and biodiesel production. Both yielded higher quantities and quality of biofuels compared to the autotrophic cells.
3. The study demonstrates an effective process that combines biotechnology and engineering to optimize microalgae for high-yield biofuel production, which could have great commercial potential for liquid fuel production.
Textile bleaching is one of the stages in the manufacture of textiles. All raw textile materials, when they are in natural form, are known as 'greige' material. This greige material will have its natural color, odor and impurities that are not suitable for clothing materials
Spps and its applications for bioactive peptides(Rajendra Sonawane)Rajendra Sonawane
Bioactive peptides are protein fragments which have a positive impact on the functions and conditions of
living beings. Peptides have shown several useful properties for human health, including antimicrobial, antifungal, antiviral,
and antitumor activities.
Determination of Anions by Ion Chromatography
1 SCOPE AND FIELD OF APPLICATION
This method is suitable for the determination of inorganic anions in Ammonia Solution in the range 100 ppb to 50 ppm m/v.
2 PRINCIPLE
The sample is passed through a column of anion exchange resin, on which the anions are absorbed and separated. They are then eluted with dilute sodium carbonate/sodium hydrogen carbonate solution and passed through a suppressor. This replaces the cations with hydrogen ions and thus reduces the background conductivity of the eluent. Final measurement is by conductivity
BENFIELD LIQUOR: DETERMINATION OF IRON
SCOPE AND FIELD OF APPLICATION
This method is suitable for the determination of the total iron in Benfield liquor samples up to a concentration of approximately 100 ppm m/v.
PerkinElmer: Environmental Contaminants in Finished Drinking Water and Raw So...PerkinElmer, Inc.
Environmental quality issues are extremely demanding, heterogeneous and ever expanding. Regulatory agencies around the world are constantly increasing the amount of environmental testing requirement to ensure public health and safety.Carbonyl compounds may be formed in water during ozonization and chlorination of natural organic matter. These, hazardous pollutants released from diverse sources including motor vehicles and industrial emissions, have been shown to have adverse effects on human health. EPA method 556 addresses this issue of carbonyl compounds in detail. This method applies to 15 carbonyl compounds.
The compounds are derivatised using pentafluoro benzyl hydroxylamine and determined on Gas chromatograph equipped with an Electron Capture detector. This GC-ECD method enables the separation, detection and quantitation of parts per billion (ppb) concentrations of low molecular weight carbonyls in water samples, safeguarding human health and ensuring compliance with industry regulations.
This method is a gas chromatographic method optimized to determine the carbonyl compounds in drinking water and raw source water. The analytes are derivatised to their corresponding penta fluorobenzyl oximes, which are extracted from water with hexane. The hexane extracts are then analyzed by GC-ECD. A PerkinElmer Elite -5 (30 meter, 0.53 mm i.d., 0.5 µm df) was used for in the method at a flow rate of 3.5 ml/min helium at constant flow mode. The oven temperature was programmed to separate the aldehyde oximes. The method is simple, fast and reproducible. The micro extraction procedure is simple and uses very small quantity of solvents which greatly reduces waste management steps and prevents pollution.
Biofilms are difficult to detect using traditional methods like culturing and can evade detection. A new method called fouling cell analysis directly detects biofilms using infrared spectroscopy to analyze the biofilm exopolymer without removing it from surfaces. Fouling cell analysis can track changes in biofilm chemistry over time and monitor the impact of cleaning methods by measuring reductions in infrared absorption peaks. Biofilms become highly resistant to cleaning and can survive harsh conditions like high temperatures and disinfectants, highlighting the need for direct biofilm detection methods.
Reactor Arrangement for Continuous Vapor Phase ChlorinationGerard B. Hawkins
Reactor Arrangement for Continuous Vapor Phase Chlorination
CONTENTS
1 BACKGROUND
2 REACTOR
3 CHEMICAL SYSTEM
4 PROCESS CHEMISTRY
5 KINETICS EXPERIMENTS AND MODELING
6 INTERPRETATION OF KINETICS INFORMATION
7 OPERATING CONDITIONS AND REACTOR DESIGN
8 REACTOR STABILITY AND CONTROL
FIGURES
1 POSTULATED REACTION PATHS FOR PROGRESSIVE CHLORINATION OF B-PICOLINE 3
2 CHLORINATION OF b-PICOLINE: MODEL PREDICTIONS OF PRODUCT DISTRIBUTION IN FULLY-MIXED REACTOR
3 TWO-STAGE REACTOR: RATE OF CHLORINATION OF b-PICOLINE
DOCUMENTS REFERRED TO IN THIS PROCESS ENGINEERING GUIDE
This document summarizes research on two proteins - Human Retinol Binding Protein-4 (RBP4) and Kvβ2, a subunit of the Kv1 potassium channel. For RBP4, the researcher optimized PCR conditions to amplify the gene and assessed RBP4's proposed role in insulin resistance and diabetes. For Kvβ2, the researcher overexpressed and purified the protein, then tested the inhibitory effects of rutin, quercetin, and resveratrol on Kvβ2 activity. The results provide new insights into physiological processes involving the Shaker potassium channel and identify resveratrol as a potential inhibitor of Kvβ2 activity. Overall, the research yielded beneficial PCR guidelines
This document is the national standard of the People's Republic of China GB 5461-2000 regarding edible salt. It revises and replaces the previous 1992 standard. The key changes include adding calculations for whiteness and chemical index based on wet percentage, stipulating sanitation index limitations, and deleting grade III products for some salt types. It specifies technical requirements, tests, quality rules, and packaging/storage for refined, washed, and solar edible salts. The main drafting units were the National Sea-lake Salt Standardization Center and China Well and Rock Salt Standardization Center.
The document summarizes information about MIOX and its Mixed Oxidant Solution (MOS) technology. In 3 sentences: MIOX produces MOS through an electrolytic process that generates hypochlorite and hydrogen peroxide from salt water. MOS is a more effective disinfectant than hypochlorite alone, able to destroy biofilms and reduce disinfection byproducts. MIOX has installed over 2,000 units across 30+ countries treating water, wastewater, pools and other applications.
This study evaluated the effect of different treatments on cobalt catalysts for 1-butene oligomerization. Sodium hydroxide (NaOH) and ammonium hydroxide (NH4OH) treatments were tested at various concentrations and with single or double treatments. Washing steps were also evaluated. The results showed that NaOH treatment achieved similar activity to NH4OH when concentrations were equal. However, double NH4OH treatment increased activity while double NaOH treatment decreased it. Washing increased activity for both treatments by potentially removing sodium ions that hindered catalysis. The study aims to better understand how treatment conditions impact cobalt catalyst performance.
This document describes the TOYOPEARL AF-rProtein A HC-650F resin for purification of monoclonal antibodies. It has the highest dynamic binding capacity of commercial protein A resins, with 70 g/L capacity for IgG at 5 minutes residence time. The recombinant protein A ligand is alkali resistant, allowing over 200 cleaning-in-place cycles with 0.1M NaOH. Testing on a therapeutic monoclonal antibody showed binding capacities remained close to 50 mg/mL even at short residence times of 45 seconds.
The document discusses various methods for developing analytical separation methods for enantiomeric mixtures using chiral chromatography columns. It covers choosing between liquid chromatography and supercritical fluid chromatography techniques, selecting appropriate chiral stationary phases, optimizing mobile phase compositions, and evaluating different columns to achieve resolution of enantiomers.
The document provides an overview of liquid-liquid extraction processes. It discusses key factors in solvent selection including selectivity, distribution coefficient, density, viscosity, and stability. Common equipment for multistage extraction includes mixer settlers, packed columns, plate columns, and mechanically agitated columns. Design considerations include flow configuration, mass transfer properties, and phase separation.
Dr. Elke Prohaska & Regina Römling BioInnovation Leader Summit TosohGBX Summits
Improving Process Efficiency in Biomanufacturing
Dr. Elke Prohaska & Regina Römling BioInnovation Leader Summit
Bench And See the Improvements at BioInnovation 2015
Similar to Cell Culture Techniques and Best Practices (20)
Hot-start DNA polymerases are commonly used in PCR for genotyping, sequencing, molecular diagnostics, and high-throughput applications. In this presentation, PCR performance of Invitrogen™ Platinum II Taq Hot-Start DNA Polymerase and Invitrogen™ AccuPrime Taq DNA Polymerase is compared in the following areas:
• PCR run time for targets of different lengths
• Amplification of AT-rich and GC-rich sequences
• Tolerance to PCR inhibitors
• Sensitivity in target detection
• Universal protocol for PCR targets of different lengths
• Multiplex PCR of 15 targets
• Product format for direct gel loading
Request a sample of Platinum II Taq enzyme at http://bit.ly/2M4U9cw
Find other PCR enzymes at http://bit.ly/2JIPrzj
Learn more about PCR at http://bit.ly/2y2aSVo
#PCR #PCREducation #Invitrogen #InvitrogenSchoolofMolBio
Human cytomegalovirus (CMV) is a common immune-evasive herpes family virus leading to lifelong asymptomatic infection in 50 to 80% of humans. Current research evaluating the use of
TCR sequencing to predict response to immunotherapy has focused on measurements of T cell clonal expansion and TCR convergence (2,3,4) as potential predictive biomarkers for
response. Given that CMV infection has been reported to elicit large clonal proliferations of CMV reactive T cells (1), and is a source of chronic antigen stimulation, we hypothesized that CMV
infection might alter T cell repertoire features in a manner relevant to the potential biomarker use of TCR sequencing. Here we sought to identify features of CMV infection using TCRB profiling of
peripheral blood (PBL) total RNA. We identify reduced T cell evenness and elevated TCR convergence as features of chronic CMV infection.
Improvement of TMB Measurement by removal of Deaminated Bases in FFPE DNAThermo Fisher Scientific
Tumor mutational burden (TMB) is a positive predictive factor for response to immune-checkpoint inhibitors in certain types of cancer. The Oncomine™ Tumor Mutation Load Assay, a targeted next generation sequencing (NGS) assay, measures TMB (from 1.2Mb of coding region) and detects mutations in 409 cancer genes. The TMB values obtained using targeted sequencing are highly correlated with TMB measured by whole exome sequencing. FFPE preservation methods can lead to significant cytosine deamination of the isolated DNA, resulting in decreased sequencing quality. In these samples, uracils are propagated as thymines and result in false C>T substitutions. Analysis of the Oncomine™ TML Assay using Torrent Suite and Ion Reporter ™ software uniquely estimates the degree of deamination in fixed tissues by measuring C:G>T:A variants. This deamination score is used to assess quality of DNA extracted from FFPE tumor tissue. To minimize the influence
that excess deamination has on TMB results, we have incorporated a repair treatment to eliminate damaged targets and improve usable TMB values of DNA from damaged FFPE tumor tissue using Uracil-DNA glycosylase (UDG). The
Oncomine™ TML Assay for TMB on the Ion Gene Studio™ S5 systems in conjunction with a deamination score is informative and potentially predictive for the use of checkpoint inhibitors in multiple cancer types.
What can we learn from oncologists? A survey of molecular testing patternsThermo Fisher Scientific
Oncologists are increasingly incorporating NGS testing to guide targeted and immuno-oncology therapies1. Most clinical NGS testing is confined to large academic institutions and reference labs, despite the fact that most cancer patients are treated in the community settings. We therefore sought to examine molecular testing selection patterns directly from oncologists in order to better identify perceived gaps in testing and treatment paradigms
Evaluation of ctDNA extraction methods and amplifiable copy number yield usin...Thermo Fisher Scientific
The use of cell-free circulating tumor DNA (ctDNA) for non-invasive cancer testing has the potential to revolutionize the field. However, emergence of an increasing number of extraction methods and detection assays is rendering laboratory workflow development much more complex and cumbersome. The use of standardized, well characterized ctDNA control materials in human plasma could facilitate the evaluation of extraction efficiency and assay performance across platforms. In this study, we use a full process ctDNA quality control material in true human plasma to demonstrate the variability of extraction yield between different ctDNA extraction kits. We also examine the correlation between the amplifiable
copy number and DNA concentration post-extraction.
Analytical Validation of the Oncomine™ Comprehensive Assay v3 with FFPE and C...Thermo Fisher Scientific
The document summarizes an analytical validation of the Oncomine Comprehensive Assay v3 (OCAv3) targeted next-generation sequencing panel performed in a CLIA-certified laboratory. The validation assessed analytical sensitivity, specificity, accuracy, and precision using formalin-fixed paraffin-embedded tumor samples and cell lines. Results showed the assay met performance thresholds of 90% or higher for detecting single nucleotide variants, insertions/deletions, copy number variants, and gene fusions across a wide range of variants. Over 2,500 clinical samples were subsequently sequenced with the assay maintaining a 95% success rate and average turnaround time of 10 days.
Novel Spatial Multiplex Screening of Uropathogens Associated with Urinary Tra...Thermo Fisher Scientific
Accurate identification of uropathogens in a timely manner is important to correctly understand urinary tract infections(UTI’s), which affects nearly 150 million people each year. The
current standard approach for detecting the UTI pathogens is culture based. This method is time consuming, has low throughput, and can lack sensitivity and/or specificity. In addition, not all uropathogens grow equally well under standard culture conditions which can result in a failure to detect the species. To address these gaps, we have developed a unique workflow from sample preparation to target identification using the nanofluidic OpenArray™ platform for spatial multiplexing of target specific assays. In this study, we tested pre-determined blinded research samples and confirmed the subset of results with orthogonal Sanger sequences.
Liquid biopsy quality control – the importance of plasma quality, sample prep...Thermo Fisher Scientific
Liquid biopsy is emerging as a non-invasive companion to traditional solid tumor biopsies. As next generation sequencing (NGS) of circulating cell-free nucleic acids (cfNA = cfDNA and cfRNA) becomes common, it’s important to understand the impact of sample preparation on quality, specificity, and sensitivity of liquid biopsy tests. Plasma samples are often limited, and may have undesirable characteristics such as lipemia or hemolysis that contribute unwanted genomic DNA (gDNA) to the sample. Low cfDNA concentration can also limit the amount available for NGS library prep. In this study, we explore the effects of suboptimal plasma and low library input on liquid biopsy NGS, and discuss various techniques for in-process quality control of cfNA samples isolated from plasma
Streamlined next generation sequencing assay development using a highly multi...Thermo Fisher Scientific
Next generation sequencing (NGS) assay development for solid tumor sequencing requires characterization of variant calling directly from formalin-fixed paraffin embedded (FFPE) tissue samples. However, cell line based FFPE and human FFPE samples only contain 2 to 20 variants, which require laboratories to invest significant resources in sample sourcing and preparation when developing assays to detect 100+ variants
Targeted T-cell receptor beta immune repertoire sequencing in several FFPE ti...Thermo Fisher Scientific
T-cell receptor beta (TCRβ) immune repertoire analysis by next-generation sequencing is a valuable tool for studies of the tumor microenvironment and potential immune responses to cancer immunotherapy. Here we describe a TCRβ sequencing assay that leverages the low sample input requirements of AmpliSeq library preparation technology to extend the capability of targeted immune repertoire sequencing to include FFPE samples which can often be degraded and in short supply
Development of Quality Control Materials for Characterization of Comprehensiv...Thermo Fisher Scientific
Targeted next-generation sequencing (NGS) panels can detect hundreds of mutations in key genes using amplification based and hybrid-capture based NGS technologies. Although NGS technology is a powerful tool, optimizing and characterizing test performance on hundreds of variants is extremely challenging, time consuming, and expensive. Samples must be sourced, variants identified and orthogonally confirmed, then quantified and diluted. This effort is then multiplied across dozens of samples, and then samples must be run over many runs and days to assess assay reproducibility, precision, sensitivity, etc. In this study, we developed a novel reference material, experimental design, and analysis pipeline that allows for highly streamlined NGS assay characterization, enabling thorough test characterization across 500+ variants within only 6 runs.
A panel was developed using the OpenArray platform to profile common respiratory tract pathogens via PCR. Assays were designed to target viral and bacterial sequences with high specificity and strain coverage. The panel demonstrated high specificity when tested against genomic standards. Pre-amplification improved sensitivity by enhancing detection of low copy targets. The panel provides a customizable and high-throughput tool for respiratory infection research.
A high-throughput approach for multi-omic testing for prostate cancer researchThermo Fisher Scientific
The proliferation of genetic testing technologies and genome-scale studies has increased our understanding of the genetic basis of complex diseases. However, this information alone tells an incomplete story of the underlying biology. Integrative approaches that combine data from multiple sources, such as the genome, transcriptome and/or proteome, can provide a more comprehensive and multi-dimensional model of complex diseases. Similarly, the integration of multiple data types in disease screening can improve our understanding of disease in populations. In a series of groundbreaking multi-omic, population-based studies of prostate cancer, researchers at the Karolinska Institutet in Stockholm, Sweden identified sets of genetic and protein biomarkers that when evaluated together with other clinical research data performed significantly better in predicting cancer risk (1,2) than the most-widely used single protein biomarker, the prostate-specific antigen (PSA).
Discover the innovations and more that led to amazing discoveries through the use of thermal cyclers. What were scientists able to accomplish? What things are important to them when selecting a thermal cycler? What do you need to advance your science?
Learn more about thermal cyclers: http://bit.ly/2Q2oPhF
See all thermal cycler offerings: http://bit.ly/2Paf1wH
A rapid library preparation method with custom assay designs for detection of...Thermo Fisher Scientific
Herein, we describe a new research method for library
preparation using the Ion AmpliSeq™ HD Library Kit with
custom assay designs from Ion AmpliSeq HD Panels for
detection of low level variants from liquid biopsy samples. This
method includes incorporation of molecular tags that enable
0.1% Limit of Detection (LOD) in cell free DNA (cfDNA) and
dual barcodes for sample identification. This method is also
applicable to formalin-fixed paraffin embedded (FFPE)
samples. The libraries can be prepared in as little as 3 hours
and are compatible for analysis with the Ion GeneStudio™ S5
system
Generation of Clonal CRISPR/Cas9-edited Human iPSC Derived Cellular Models an...Thermo Fisher Scientific
This document describes a workflow for generating clonal CRISPR-edited human induced pluripotent stem cell (hiPSC) lines. Key aspects of the workflow include:
1) Developing a hiPSC line that stably expresses Cas9 to facilitate efficient genome editing.
2) Optimizing delivery methods for CRISPR/Cas9 editing tools and improving single cell clone survival and isolation using laminin-521 and StemFlex medium.
3) Applying the workflow to generate hiPSC lines carrying disease-relevant mutations and testing the cell models in assays, finding increased sensitivity to stress in models of Parkinson's disease.
TaqMan®Advanced miRNA cDNA synthesis kit to simultaneously study expression o...Thermo Fisher Scientific
MicroRNAs (miRNA) are a class of small non-coding RNAs (approximately 21 nt long) that bind complementary sequences in target mRNAs to specifically regulate gene expression. Aberrant regulation of miRNAs and their targets has been associated with several diseases including cancer. The relationship between miRNA and mRNA has been found to be important in cancer development and progression. Simultaneous expression studies of miRNA and mRNA and detection of mutations in mRNA transcripts can be valuable in understanding molecular mechanisms that
have an underlying role in various diseases. We demonstrate the technical verification of a novel method to reverse-transcribe and pre-amplify miRNA and mRNA from sample-limiting serum research samples using the TaqMan® Advanced miRNA cDNA Synthesis Kit. Based on results from previous studies, a signature of 49 mRNA and 37 miRNA targets has been identified that may help distinguish between benign and malignant pancreatic tissues. In this study, these targets and an additional set of transcript mutations were analyzed in serum from normal and test samples. TaqMan assays for miRNA and mRNA targets and custom TaqMan Mutation Detection Assays (TMDAs) were placed on TaqMan Array Cards to facilitate investigation of several samples in a single experiment. Results demonstrate that transcript mutations can be detected and miRNA and mRNA targets can be reliably quantified from a single reverse transcription reaction. For research use only. Not for use in diagnostic purposes.
Identifying novel and druggable targets in a triple negative breast cancer ce...Thermo Fisher Scientific
In this study, we developed a CRISPR/Cas9-based high throughput loss-of-function screen for identifying target genes responsible for the tumor proliferation and growth in TNBC. Our initial focus was to identify essential kinases in MDA-MB-231 cell line using the Invitrogen™ LentiArray™ Human Kinase CRISPR Library, which targets 840 kinases with up to 4 different gRNAs per protein kinase for complete gene knockout. This functional screen identified over 90 protein kinases that are essential for cell viability and cell proliferation. Ten of these hits (CDK1, CDK2, CDK8, CDK10, CDK11A, CDK19, CDK19, CDC7, EPHA2 and WEE1) are well-known targets validated in the literature. Currently, we are in the process validating the novel hits through target gene sequencing, western blotting and target specific small molecule kinase inhibitors.
Evidence for antigen-driven TCRβ chain convergence in the melanoma-infiltrati...Thermo Fisher Scientific
T cell convergence refers to the phenomenon whereby antigen-driven selection enriches for T cell receptors (TCRs) having a shared antigen specificity but different amino acid or
nucleotide sequence. T cell recruitment and expansion within the tumor microenvironment (TME) may be directed by responses to tumor neoantigen, suggesting that elevated T
cell convergence could be a general feature of the tumor infiltrating T cell repertoire. Here we use the Ion AmpliSeq™ Immune Repertoire Assay Plus – TCRβ to evaluate evidence
for T cell convergence within melanoma tumor biopsy research samples from a set of 63 subjects plus peripheral blood leukocytes (PBL) from four healthy subjects. We find that the melanoma TME is highly enriched for convergent TCRs compared to healthy donor peripheral blood. We discuss the potential use of TCR convergence as a liquid biopsy compatible predictive biomarker for immunotherapy response.
Analytical performance of a novel next generation sequencing assay for Myeloi...Thermo Fisher Scientific
To support clinical and translational research into precision oncology strategies for myeloid cancers, a next-generation sequencing (NGS) assay was developed to detect common and relevant somatic alterations. To define gene targets that were recurrently altered in myeloid cancers and relevant for clinical and translational research, an extensive survey of investigators at hematology oncology research labs was performed.
A workshop hosted by the South African Journal of Science aimed at postgraduate students and early career researchers with little or no experience in writing and publishing journal articles.
हिंदी वर्णमाला पीपीटी, hindi alphabet PPT presentation, hindi varnamala PPT, Hindi Varnamala pdf, हिंदी स्वर, हिंदी व्यंजन, sikhiye hindi varnmala, dr. mulla adam ali, hindi language and literature, hindi alphabet with drawing, hindi alphabet pdf, hindi varnamala for childrens, hindi language, hindi varnamala practice for kids, https://www.drmullaadamali.com
This slide is special for master students (MIBS & MIFB) in UUM. Also useful for readers who are interested in the topic of contemporary Islamic banking.
Main Java[All of the Base Concepts}.docxadhitya5119
This is part 1 of my Java Learning Journey. This Contains Custom methods, classes, constructors, packages, multithreading , try- catch block, finally block and more.
How to Fix the Import Error in the Odoo 17Celine George
An import error occurs when a program fails to import a module or library, disrupting its execution. In languages like Python, this issue arises when the specified module cannot be found or accessed, hindering the program's functionality. Resolving import errors is crucial for maintaining smooth software operation and uninterrupted development processes.
Walmart Business+ and Spark Good for Nonprofits.pdfTechSoup
"Learn about all the ways Walmart supports nonprofit organizations.
You will hear from Liz Willett, the Head of Nonprofits, and hear about what Walmart is doing to help nonprofits, including Walmart Business and Spark Good. Walmart Business+ is a new offer for nonprofits that offers discounts and also streamlines nonprofits order and expense tracking, saving time and money.
The webinar may also give some examples on how nonprofits can best leverage Walmart Business+.
The event will cover the following::
Walmart Business + (https://business.walmart.com/plus) is a new shopping experience for nonprofits, schools, and local business customers that connects an exclusive online shopping experience to stores. Benefits include free delivery and shipping, a 'Spend Analytics” feature, special discounts, deals and tax-exempt shopping.
Special TechSoup offer for a free 180 days membership, and up to $150 in discounts on eligible orders.
Spark Good (walmart.com/sparkgood) is a charitable platform that enables nonprofits to receive donations directly from customers and associates.
Answers about how you can do more with Walmart!"
How to Manage Your Lost Opportunities in Odoo 17 CRMCeline George
Odoo 17 CRM allows us to track why we lose sales opportunities with "Lost Reasons." This helps analyze our sales process and identify areas for improvement. Here's how to configure lost reasons in Odoo 17 CRM
বাংলাদেশের অর্থনৈতিক সমীক্ষা ২০২৪ [Bangladesh Economic Review 2024 Bangla.pdf] কম্পিউটার , ট্যাব ও স্মার্ট ফোন ভার্সন সহ সম্পূর্ণ বাংলা ই-বুক বা pdf বই " সুচিপত্র ...বুকমার্ক মেনু 🔖 ও হাইপার লিংক মেনু 📝👆 যুক্ত ..
আমাদের সবার জন্য খুব খুব গুরুত্বপূর্ণ একটি বই ..বিসিএস, ব্যাংক, ইউনিভার্সিটি ভর্তি ও যে কোন প্রতিযোগিতা মূলক পরীক্ষার জন্য এর খুব ইম্পরট্যান্ট একটি বিষয় ...তাছাড়া বাংলাদেশের সাম্প্রতিক যে কোন ডাটা বা তথ্য এই বইতে পাবেন ...
তাই একজন নাগরিক হিসাবে এই তথ্য গুলো আপনার জানা প্রয়োজন ...।
বিসিএস ও ব্যাংক এর লিখিত পরীক্ষা ...+এছাড়া মাধ্যমিক ও উচ্চমাধ্যমিকের স্টুডেন্টদের জন্য অনেক কাজে আসবে ...
LAND USE LAND COVER AND NDVI OF MIRZAPUR DISTRICT, UPRAHUL
This Dissertation explores the particular circumstances of Mirzapur, a region located in the
core of India. Mirzapur, with its varied terrains and abundant biodiversity, offers an optimal
environment for investigating the changes in vegetation cover dynamics. Our study utilizes
advanced technologies such as GIS (Geographic Information Systems) and Remote sensing to
analyze the transformations that have taken place over the course of a decade.
The complex relationship between human activities and the environment has been the focus
of extensive research and worry. As the global community grapples with swift urbanization,
population expansion, and economic progress, the effects on natural ecosystems are becoming
more evident. A crucial element of this impact is the alteration of vegetation cover, which plays a
significant role in maintaining the ecological equilibrium of our planet.Land serves as the foundation for all human activities and provides the necessary materials for
these activities. As the most crucial natural resource, its utilization by humans results in different
'Land uses,' which are determined by both human activities and the physical characteristics of the
land.
The utilization of land is impacted by human needs and environmental factors. In countries
like India, rapid population growth and the emphasis on extensive resource exploitation can lead
to significant land degradation, adversely affecting the region's land cover.
Therefore, human intervention has significantly influenced land use patterns over many
centuries, evolving its structure over time and space. In the present era, these changes have
accelerated due to factors such as agriculture and urbanization. Information regarding land use and
cover is essential for various planning and management tasks related to the Earth's surface,
providing crucial environmental data for scientific, resource management, policy purposes, and
diverse human activities.
Accurate understanding of land use and cover is imperative for the development planning
of any area. Consequently, a wide range of professionals, including earth system scientists, land
and water managers, and urban planners, are interested in obtaining data on land use and cover
changes, conversion trends, and other related patterns. The spatial dimensions of land use and
cover support policymakers and scientists in making well-informed decisions, as alterations in
these patterns indicate shifts in economic and social conditions. Monitoring such changes with the
help of Advanced technologies like Remote Sensing and Geographic Information Systems is
crucial for coordinated efforts across different administrative levels. Advanced technologies like
Remote Sensing and Geographic Information Systems
9
Changes in vegetation cover refer to variations in the distribution, composition, and overall
structure of plant communities across different temporal and spatial scales. These changes can
occur natural.
LAND USE LAND COVER AND NDVI OF MIRZAPUR DISTRICT, UP
Cell Culture Techniques and Best Practices
1. Watch the webinar
Cell Culture Best Practices
Busting Myths & Improving Outcomes
Gibco® Cell Culture
and
Timothy Fawcett, Ph.D.
BioSciConcepts/BioTechnical Institute of Maryland, Inc.
2. Agenda Overview
Understanding the in-vitro system,
Working with components of cell culture media & formulations, serum
performance characteristics,
Aspects of cell culture productivity, inter-relationships between cell
growth cycle & its environment,
Tips & techniques for maximizing recovery……and along the way we will
challenge lab-lores and bust some myths!
2
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3. 3
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4. Guidance on Good Cell Culture
Guidance has been prepared to promote an awareness of a
broad range of important issues in cell culture in workers
coming to use cells for their work for the first time, and to
remind others of the fundamental aspects of good practice in
cell and tissue culture.
Based on a report of the second European Centre for the Validation of Alternative Methods task force on
Good Cell Culture Practice; Coeche, et al, ATLA 33, 261-287, 2005.
4
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5. GCCP Guidance
Based Upon Six Operational Principles:
1. Establishment and maintenance of a sufficient understanding
of the in vitro system and of the relevant factors which could
affect it.
2. Assurance of the quality of all materials and
methods…reproducibility.
3. Documentation
4. Establishment and maintenance of adequate measures to
protect individuals and the environment from any potential
hazards.
5. Compliance with relevant laws and regulations, and with
ethical principles.
6. Provision of relevant and adequate education and training for
all personnel, to provide high quality work and safety.
5
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6. BHK-21 Confluent?
6
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7. Hepatocytes ?
7
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8. Will This Result in a Culture Change?
8
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9. Cell Culture Media:
Components & Characteristics
9
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10. Cell Culture: Media, Reagents
Cell culture media is a Components Commonly Added
to Culture Media
mixture of components
used to simulate the
natural environment of Inorganic Salts
the cell. − Bulk Ion Salts and Trace Elements
Amino Acids
− Essential and Non-Essential
Vitamins
Sugars
Phenol Red
10
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11. Media Components & Characteristics
Cell culture media helps maintain correct osmotic pressure and
ionic equilibrium.
Cell culture media must provide buffering capacity.
pH needs to be maintained at pH=7.4
• The salts and amino acids in media provide some buffering capacity.
• H2CO2/NaH2CO3 system requires CO2 to maintain pH.
• If no CO2 is available use HEPES or NaH2PO4 buffer system.
11
Watch this as a webinar 5/24/2012 | Life Technologies™ Proprietary and confidential
13. Media Components & Characteristics
Glutamine, a major catabolite for fast growing cells-Source of ATP
Short half-life: 3 weeks at 4oC, 5 days at 37oC
The rate of decomposition is pH dependent and temperature
dependent
In rapidly growing cultures, glutamine levels can go to zero in 48
hours
Media
serves as a carbon source for purine biosynthesis.
is a donor of primary amino groups, for the synthesis of pyrimidines and
asparagine
13
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14. MYTH…Adding more Glutamine is Good
+ L-glutamine
+ FBS
Efficient energy metabolism
High growth yield
GlutaMAX™ is better!
Ammonia &
pryyolidine
carboxylic
acid
14
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15. Adding GlutaMAX™ is BETTER
• L-glutamine and GlutaMAX supplemented into DMEM
• Left at 37⁰ C for 7 days
• Ammonia measured each day via HPLC
• Glutamine breakdown was rapid compared to GlutaMAX
15
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16. Cell Culture: Water
Cell culture must be of the highest quality
with little or no bio-burden present.
− Seasonal variation in water quality can be a potential problem
− Dissolved gasses, rust inhibitors and organics can be present in water
− Water used for cell culture must be highly purified
− Usually multiple approaches to purification are taken
− Water For Injection …e.g. Gibco® WFI
> same as in the cGMP / ISO controlled manufacturing process
> saves steps, time – already triple filtered
16
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17. Cell Culture: Serum
Serum for cell culture
A biological product isolated and processed under very stringent
QC & manufacturing conditions
Provides additional nutrients and growth/attachment factors
Serum is undefined ~10,000 components
When serum has been added to basal media it is termed
COMPLETE MEDIA
Animal Sera Provides: Growth Factors, Proteins, Attachment
Factors, Hormones, Lipids and Minerals and Other Nutrients.
17
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18. Que Sera Sera…..?
Certified FBS - US FBS – Australian
Horse serum Goat serum
MYTH: All Sera is the same…..!
Lamb serum
Ultra-low IgG FBS
Qualified FBS - US
NB calf serum
Rabbit serum
FBS – New Zealand
Dialyzed FBS
MSC Qualified FBS
Charcoal Stripped FBS ESC Qualified FBS
Porcine serum
Qualified FBS–USDA, SA Chicken serum
18
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19. But we work with a broad range of cells…..
Robust /Not Sensitive Finicky/Sensitive
CHO A549 Jurkat
HeLa Adipose-derived Embryonic
MCF-7 stem cells stem cells
HEK293 3t3 Primary
Fibroblast Mesenchymal
fibroblasts
stem cells
HUVEC
Primary
Neurons
19
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20. Use a serum that meets your research needs
• Lowest BSE risk/least viral load
• Lowest endotoxin levels
• Need for biochemical & hormonal profile certification
• Sensitive, finicky cell lines or general cell culture
• Specific applications and assays, need for OBS…..?
Choose the right sera for your specific research
Not all sera is qualified and validated for the right applications
20
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21. Sera Experimental Design
• Gibco® Certified, Qualified grade FBS – various lots
• Versus a competitor
• Tested with several human cell lines
HEK293: Human Embryonic Kidney - epithelial
HeLa (Henrietta Lacks): epithelial - cervix cancer
A549: epithelial- lung
MCF-7: epithelial - breast cancer
Jurkat: lymphoblast - cancer
• Measured cell growth and viability over 5 passages
21
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22. ‘Consistency of cell growth over time & passages’ is the # 1 need
for sera - according to researchers*
A549 MCF-7
0.90 2.00
0.80 1.80
1.60
0.70
Viable Cells/mL (x106)
Viable Cells/mL (x106)
1.40
0.60
1.20
0.50 Qualified, Qualified,
USDA 1.00 USDA
0.40 Certified, US Certified, US
0.80
0.30
Competitor P 0.60 Competitor P
0.20 0.40
0.10 0.20
0.00 0.00
Passage 1 Passage 2 Passage 3 Passage 4 Passage 5 Passage 1 Passage 2 Passage 3 Passage 4 Passage 5
A549 cells grow consistently in Gibco® MCF-7 cells grow consistently in Gibco®
Certified, US Certified, US
Watch this as a webinar
22 5/24/2012 | Life Technologies™ Proprietary and confidential
* Percepta Life Sciences quantitative market research for Sera VOC Need-Gaps, Nov 2010 among US & EU researchers.
23. ‘Consistency of cell growth over time & passages’ is the # 1 need
for sera - according to researchers
HEK293
4.50
4.00
3.50
Viable Cells/mL (x106)
3.00
2.50 Qualified, USDA
2.00
Certified, US
1.50
1.00 Competitor P
0.50
0.00
Passage 1 Passage 2 Passage 3 Passage 4 Passage 5
HEK293 cells grow consistently in Gibco® USDA Qualified
23
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24. Gibco® ES Cells Qualified FBS
validated with qRT-PCR Assay
Quantitative assay from Gibco® vs. traditional methods for other brands
Improved lot-to-lot consistency of GIBCO® ESC FBS qualities
Detection in variation of differentiation and pluripotency
Fast and accurate quantitation of gene expression
24
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25. Gibco Qualified ESC FBS Performance Comparison
Expression of Pluripotency Gene
1.80
•Pluripotency Gene 1.60
Level of Pluripotency
1.40
• Transcription factor essential 1.20
for self-renewal 1.00
0.80
• Controls multiple key 0.60
transcription factors 0.40
0.20
• GIBCO® ES Cell FBS lots are 0.00
selected for high expression Control gibco ES Cell gibco Non-ES Competitor M Competitor S
FBS Qualified Cell FBS
by New Assay
Expression of Differentiation Gene
2.50
•Differentiation Gene
Level of Differentiation
2.00
• Expressed at early stages of 1.50
development 1.00
• Indicator of early differentiation 0.50
• GIBCO® ES Cell FBS lots are 0.00
selected for low expression Control gibco ES Cell gibco Non-ES Competitor M Competitor S
FBS Qualified Cell FBS
by New Assay
25
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26. Sera: Lab-Lores ?
Concerns about variability when using FBS
− Unique Lot Matching Service for Gibco® Sera: Gibco® iMATCH™
> proprietary algorithm based on performance & test variables to offer closest
match to previous lots
> check www.invitrogen.com/imatch
Concerns about Flocculence
− Only a coagulation of proteins, look like strings, not abnormal
Sera (or Media) - cannot be sterile
− manufactured & tested for sterility
FBS will influence or differentiate stem cells
− ES Cell Qualified FBS validated with qRT-PCR assay from Gibco®
26
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27. Cell Culture Media:
Effect Of Environmental Factors
27
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28. Effect of Light on Media Performance
120%
100%
80%
60%
Light
40% No Light
20%
0%
0.5 2.25 4.25 6.25
hours hours hours hours
Sp2 Growth as a Percent of Control
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29. Critical Parameters for Optimal Media Performance
Recognize there is a natural decay rate of unstable compounds
in media
Maintain correct pH, affects the stability of some compounds Qu i c k T i m e ™ a n d a
d e c o m p re s s o r
a re n e e d e d t o s e e t h i s p i c t u re .
Freeze serum only one time
Do not freeze basal media
PROTECT FROM LIGHT
Minimize the time media is at 37oC.
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30. “Trypsinization”, Detachment of Adherent Cells
Trypsin
− Traditional, contains animal products.
− Cleaves arg and lys amino acids
− Needs to be quenched using serum or soybean trypsin inhibitor
− A mixture of enzymes
− Degrades itself
TrypLE™ Express
− A recombinant fungal trypsin-like enzyme
− High purity
− Lower cell toxicity
− Room temperature stable
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31. MYTH – Nothing’s as Good as Trypsin to Detach
Adherent Cells
• Compared TrypLE™ Select 10x with Trypsin and other cell dissociation products
• Tested with 5 strongly adherent cell types
Mean Time Required for Cell Release
40
Trypsin-EDTA
35 1x TrypLE
Time for cell release (Minute)
1x TrypLE Diluted
30
10x TrypLE
25 HyQTase
Accutase
20 TrypZean
15
10
5
0
MDCK PK-15 A549 MSC SFM MSC+10%FBS
Cell Lines
TrypLE™ Select 10x – superior performance
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32. Using TrypLE™ Select 10x is BETTER
• Removing cells from plastic surface is ROUGH on cells
• TrypLE™ Select 1x or 10x is GENTLE
• TrypLE™ Select 1x or 10x is AOF
• TrypLE™ Select 1x or 10x is STABLE at room temperature
• Can be used on ALL CELL TYPES
32
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33. Cell Culture:
Tips & Tricks
33
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34. Tips: Freezing and Thawing
Freeze cells slowly, 1oC /minute.
Thaw as rapidly as possible to 37oC.
− Use a cup of 37oC water to move the vial of cells from freezer to 37oC for
rapid thawing.
− Place cells SLOWLY into pre-warmed media.
Freeze early passage, healthy, well fed cells.
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35. Tips: Freezing & Thawing
Use Conditioned Media
− Conditioned media is media that has been used to grow cells.
− Has unknown factors from paracrine growth.
− Great for getting cells through difficult times or when growing cells at very low density.
Use a Styrofoam tray when organizing cells
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36. Tips: Efficiency & Reduced Risk of Contamination
Aliquoting sera from 500 ml or liter bottles
− Will a 50 ml easy-to-use bottle be efficient…?
− Consider Gibco® 50ml One Shot™ FBS bottles
> easy to use, quicker to thaw
> reduced risk of contamination
Also consider using Lab Armor™ Beads with Gibco® products
− efficient and reduced contamination risk compared to water baths
+ = Peace of Mind!
50 ml One Shot™ FBS Lab Armor™ Beads
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37. Resources & Sampling Offers
Glutamax www.lifetechnologies.com/glutamax
TrypLE www.lifetechnologies.com/trypLE
Gibco FBS Samples www.lifetechnologies.com/fbs
Gibco ES Cell FBS www.lifetechnologies.com/escfbs
Cell Culture Basics Videos www.lifetechnologies.com/cellculturebasics
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38. THANK YOU
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