This document provides an overview of histology and tissue preparation techniques. It discusses the following key points:
- Histology is the study of tissues and how cells are organized into tissues and organs. The four main types of tissues are epithelial, connective, nervous, and muscular tissue.
- Tissue samples are obtained through biopsies and prepared through a process including fixation, dehydration, clearing, embedding in paraffin wax or resin, sectioning with a microtome, and staining. Special staining techniques can identify structures like glycogen or calcium.
- Histochemistry uses chemical reactions to identify certain molecules in tissues, while immunohistochemistry uses labeled antibodies to identify antigens within tissues under a microscope.
Whole mount preservation involves mounting small, thin specimens whole without sectioning. This preserves the natural shape, color and structure. Methods include Hygrobutol for algae, Dioxan or Creosote for transparency, and glycerin-xylol or Venetian turpentine for dehydration before mounting in balsam or turpentine. Temporary mounts use glycerin or lactophenol, while semi-permanent mounts use glycerine jelly which is filtered gelatin with glycerin and phenol. Whole mounting reveals details not seen in sections and preserves the natural appearance of specimens like algae and fungi.
Introduction to organ culture in plant tissue culture and root cultureCollege
This presentation is all about the organ culture and its applications which is an important aspect in Plant tissue culture today. Also this presentation provide detail information about root culture and its basic appilication
This document discusses several major plant hormones (phytohormones), including their functions, locations of synthesis, and some uses. It describes auxins, cytokinins, gibberellins, abscisic acid, and ethylene - noting that they regulate growth, stress responses, germination, flowering, and fruit development. Specific hormones are discussed in more detail, outlining their natural and synthetic forms as well as some of their roles and applications in agriculture.
Hello ever one i hope its useful for preparation of notes regarding plant tissue culture for Pharmacognosy .. B.pharm II yr IV sem.. plz give comments it may useful for me and i can rectify the things.
Plant tissue culture involves growing plant cells, tissues or organs in sterile conditions on nutrient media. The key hormones auxin and cytokinin play important roles in differentiation. Tissue culture applications include micropropagation, germplasm preservation, haploid production, and genetic engineering. Important techniques include somatic embryogenesis, organogenesis, microcutting, anther/microspore culture, protoplast culture, and callus and cell suspension culture.
Essay on Plant Tissue Culture Contents:
the Definition of Plant Tissue Culture.
the History of Plant Tissue Culture.
the Basic Requirements of Plant Tissue Culture.
the General Techniques of Plant Tissue Culture.
the Basic Aspects of Plant Tissue Culture.
the Cellular Totipotency.
the Differentiation.
the Methods in Plant Tissue Culture.
the Applications of Plant Tissue Culture.
the Morphogenesis.
the Subculture or Secondary Cell Culture.
the Soma-Clonal Variation.
the Somatic Hybrids and Cybrids.
the Micro-Propagation.
the Artificial Seed.
the Cryopreservation.
This document discusses plant tissue culture and protoplast fusion. Protoplast fusion allows for crossing plants that would otherwise be unable to cross by mixing and fusing the protoplasts, or plant cells without cell walls, from different plant species. This forms a binucleate cell containing nuclei from both parental plants, introducing new genes. The process involves removing the cell wall from plant cells using enzymes, incubating them in a salt solution, and culturing the protoplasts in a suitable medium to encourage cell wall regeneration and growth.
This document summarizes the main plant hormones: auxins, gibberellins, cytokinins, ethylene, and abscisic acid. It describes their functions in regulating plant growth processes like cell division, elongation, flowering, fruit development, senescence, and stress response. Specific examples are given to illustrate how each hormone influences these processes and commercial applications that exploit hormonal effects, such as promoting fruit ripening or inhibiting leaf abscission. The presentation concludes by asking if the audience has any questions.
Whole mount preservation involves mounting small, thin specimens whole without sectioning. This preserves the natural shape, color and structure. Methods include Hygrobutol for algae, Dioxan or Creosote for transparency, and glycerin-xylol or Venetian turpentine for dehydration before mounting in balsam or turpentine. Temporary mounts use glycerin or lactophenol, while semi-permanent mounts use glycerine jelly which is filtered gelatin with glycerin and phenol. Whole mounting reveals details not seen in sections and preserves the natural appearance of specimens like algae and fungi.
Introduction to organ culture in plant tissue culture and root cultureCollege
This presentation is all about the organ culture and its applications which is an important aspect in Plant tissue culture today. Also this presentation provide detail information about root culture and its basic appilication
This document discusses several major plant hormones (phytohormones), including their functions, locations of synthesis, and some uses. It describes auxins, cytokinins, gibberellins, abscisic acid, and ethylene - noting that they regulate growth, stress responses, germination, flowering, and fruit development. Specific hormones are discussed in more detail, outlining their natural and synthetic forms as well as some of their roles and applications in agriculture.
Hello ever one i hope its useful for preparation of notes regarding plant tissue culture for Pharmacognosy .. B.pharm II yr IV sem.. plz give comments it may useful for me and i can rectify the things.
Plant tissue culture involves growing plant cells, tissues or organs in sterile conditions on nutrient media. The key hormones auxin and cytokinin play important roles in differentiation. Tissue culture applications include micropropagation, germplasm preservation, haploid production, and genetic engineering. Important techniques include somatic embryogenesis, organogenesis, microcutting, anther/microspore culture, protoplast culture, and callus and cell suspension culture.
Essay on Plant Tissue Culture Contents:
the Definition of Plant Tissue Culture.
the History of Plant Tissue Culture.
the Basic Requirements of Plant Tissue Culture.
the General Techniques of Plant Tissue Culture.
the Basic Aspects of Plant Tissue Culture.
the Cellular Totipotency.
the Differentiation.
the Methods in Plant Tissue Culture.
the Applications of Plant Tissue Culture.
the Morphogenesis.
the Subculture or Secondary Cell Culture.
the Soma-Clonal Variation.
the Somatic Hybrids and Cybrids.
the Micro-Propagation.
the Artificial Seed.
the Cryopreservation.
This document discusses plant tissue culture and protoplast fusion. Protoplast fusion allows for crossing plants that would otherwise be unable to cross by mixing and fusing the protoplasts, or plant cells without cell walls, from different plant species. This forms a binucleate cell containing nuclei from both parental plants, introducing new genes. The process involves removing the cell wall from plant cells using enzymes, incubating them in a salt solution, and culturing the protoplasts in a suitable medium to encourage cell wall regeneration and growth.
This document summarizes the main plant hormones: auxins, gibberellins, cytokinins, ethylene, and abscisic acid. It describes their functions in regulating plant growth processes like cell division, elongation, flowering, fruit development, senescence, and stress response. Specific examples are given to illustrate how each hormone influences these processes and commercial applications that exploit hormonal effects, such as promoting fruit ripening or inhibiting leaf abscission. The presentation concludes by asking if the audience has any questions.
Cyanobacteria, also known as blue-green algae, are photosynthetic prokaryotes that reproduce through binary fission and have specialized structures like hormogones, akinetes, and heterocysts that aid in reproduction and survival. They have a multilayer cell wall and lack organelles but contain pigments like chlorophyll and phycobilins within thylakoid membranes to perform photosynthesis. Cyanobacteria inhabit various environments like freshwater, marine water, soil and hot springs.
The document discusses plant growth and development. It defines growth as a permanent increase in plant mass through cell division and enlargement. Growth occurs in phases including formation, elongation, and maturation. Development includes differentiation, where cells take on specialized structures and functions. The document also discusses leaf growth, flowering, pollination, fertilization, seed formation, and fruit development. Environmental factors and phytohormones influence growth and developmental processes.
This document discusses maceration and smear techniques. Maceration involves separating plant or animal tissues into individual cells or groups by dissolving the middle lamella. Three common maceration methods are described: Schultze's method using nitric acid and potassium chlorate, Jeffrey's method using boiling and chromic acid, and Harlow's method using chlorine water. Smears are used to study internal cell details like nuclei and are prepared by smearing material on a slide. Two staining methods for smears are acetocarmine, which stains and fixes in one step, and Feulgen's method using basic fuchsin.
MICROTECHNIQUE Killing and fixation moduleAbdulsalm
This document provides instructions for preparing biological specimens for light microscopy. It discusses the key steps of sample collection, killing and fixation, dehydration, clearing, paraffin embedding, microtomy, staining, and observation. Specific fixation fluids, dehydration reagents, clearing agents, and staining methods are described. The goal is to preserve specimens while modifying properties like refractive index to allow examination under a light microscope.
This document provides an overview of cryopreservation, which involves preserving biological material such as cells, tissues, organs, and embryos at ultra-low temperatures, typically in liquid nitrogen. It discusses the history, principles, mechanisms, and applications of cryopreservation. Key aspects covered include the use of cryoprotectants to prevent freezing damage to cells, various freezing and thawing methods, long-term storage in liquid nitrogen, and viability testing after thawing to regenerate plants or animals from preserved material. Cryopreservation has important applications in biobanking, conservation of endangered species, and preservation of disease-free agricultural crops.
Auxins are a specific group of plant hormones that play an important role in plant growth and development. Some key points about auxins include:
- They were the first plant hormone discovered. Common auxins include indole-3-acetic acid (IAA).
- Auxins are synthesized in root tips, young leaves, and apical buds, and are involved in processes like stem elongation, root initiation, apical dominance, and fruit development.
- Their discovery was made through experiments by Charles Darwin, Peter Boysen-Jensen, and Frits Went in the late 19th/early 20th centuries which demonstrated that a signal substance moved through plants and caused differential growth.
Tissue culture involves growing plant and animal cells, tissues, or organs artificially in a sterile environment with a nutrient medium. It allows for cloning and mass propagation of plants and animals. Tissue culture requires selecting appropriate explant tissue, developing a sterile culture medium with growth regulators, and regularly subculturing and multiplying the cells. It has many applications like rapidly propagating desirable crops and endangered species, providing pathogen-free plants, selecting for improved varieties, and studying cell processes. Animal tissue culture differs in requiring serum, having limited cell divisions, and posing some biohazard risks. Its uses include growing viruses, producing monoclonal antibodies, and facilitating genetic studies.
Plant tissue culture is a collection of techniques used to grow plant cells, tissues or organs under sterile conditions. It allows for the mass production of clones of plants with desirable traits. The key aspects of plant tissue culture are maintaining sterile conditions on a nutrient medium, and providing proper aeration. Common types of plant tissue culture include callus culture, single cell culture, root tip culture, shoot tip culture, and anther culture. Plant tissue culture has many applications for plant conservation, breeding, and production of secondary metabolites.
The fungal cell wall is composed of various components that give shape, strength, and protection to fungi. The main components are chitin, glucan, and proteins. Chitin is a nitrogenous polysaccharide made of N-acetylglucosamine that forms microfibrils providing structure. Glucan is a polysaccharide of glucose monomers linked by beta glycosidic bonds that is a major component of fungal cell walls. Glycoproteins and proteins are also embedded in the cell wall matrix and contain carbohydrates. Together these components form the primary wall and secondary wall layers that define fungal cells.
Fungi have several distinguishing morphological features:
1. They have cell walls containing chitin and lack peptidoglycan.
2. They can exist in both unicellular and multicellular forms, dividing asexually or sexually.
3. They are classified based on their structures - yeasts are unicellular, molds form branching hyphae and mycelium, and dimorphic fungi switch between yeast and mold forms based on temperature.
The isolation, culture and fusion of protoplasts is a fascinating field in plant research. Protoplast isolation and their cultures provide millions of single cells (comparable to microbial cells) for a variety of studies.
Vital staining involves using non-toxic dyes on living organisms and cellular structures. Common vital stains include neutral red, methylene blue, and Janus green B. Vital stains can be used to differentiate between living and dead cells, stain specific organelles like mitochondria, and study pathological cell changes without harming living tissues. Vital stains are classified based on their chemical properties and degree of dissociation into categories like basic, acidic, atmospheric and electro neutral. Techniques for vital staining include progressive, regressive, counterstaining, and double staining using combinations of contrasting dyes.
basic principles and protocol in plant tissue cultureAlia Najiha
Plant tissue culture involves growing plant cells, tissues, or organs in an artificial nutrient medium under sterile conditions. It allows for the rapid production of exact genetic copies of plants and is used to regenerate whole plants from genetically modified plant cells. The key requirements for successful tissue culture are appropriate explant tissue, a suitable growth medium, aseptic conditions to prevent microbial contamination, growth regulators, and frequent subculturing to provide nutrients and remove waste. The basic steps are selection of explant tissue from a healthy mother plant, sterilization of the tissue, establishment of the explant in a culture medium, multiplication through callus formation and shoot development, and root formation through manipulation of growth regulators. Benefits include providing pathogen-free plants
The document discusses the structure and functions of plant stems. It begins by defining stems and describing their main functions: support, conduction, and production of new living tissue. It then discusses the external morphology of stems, including nodes, internodes, buds, and other features. The internal structure is also examined, focusing on the shoot apex organization and primary meristems that give rise to primary tissues like epidermis, ground tissue, and vascular tissues. Primary growth and tissue development in dicot and monocot stems is contrasted. Finally, the document covers secondary growth, which occurs in woody stems through the activity of the vascular cambium and cork cambium.
Cryopreservation is the process of preserving living cells and tissues by cooling them to low sub-zero temperatures. This stops any chemical or enzymatic activity in the cells that could cause damage. Traditional cryopreservation relies on coating materials in cryoprotectants like glycerol or DMSO to prevent ice formation during freezing. Cryopreservation has applications for preserving reproductive cells, embryos, and tissues and is also used in the unproven field of cryonics which seeks to preserve entire human bodies for possible future revival.
This document provides instructions for sterilizing plant explants for tissue culture. It discusses that culture media supports microbial growth which can kill plant tissues, so complete sterilization is essential. The key steps outlined are: washing explants with detergent and water; submerging in disinfectants like ethanol or mercury chloride; rinsing with sterile water; and sealing cultures with sterile cotton plugs. A two-step sterilization using ethanol prior to another disinfectant can improve sterility. Proper sterilization will increase the success rate of tissue culture by reducing contamination and allowing explant growth without microbes.
Gibberellic Acid or Gibberellin Hormonesvidan biology
Gibberellins (GAs) are plant hormones that regulate various developmental processes. They are tetracyclic diterpenoid acids synthesized via the terpenoid pathway in plastids and then modified in the endoplasmic reticulum and cytosol. Bioactive GAs include GA1, GA3, GA4, and GA7 and contain a carboxyl group at C-7. GAs are synthesized in shoots and roots and translocated via phloem and xylem, respectively. DELLA proteins repress growth but are degraded by the 26S proteasome in response to GA binding to the GID1 receptor, which forms a complex targeting DELLA for ubiquitination and degradation. This releases repression
Cytokinins are a class of plant growth hormones that promote cell division. They were first discovered in 1955 from degraded herring sperm DNA. There are two main types: adenine-type cytokinins like kinetin and zeatin, and phenylurea-type cytokinins. Cytokinins play many important physiological roles in plants including cell division, cell elongation, differentiation, delaying senescence, and influencing flowering. They are widely used in tissue culture and to increase the shelf life of cut flowers and vegetables.
Plant tissues can be divided into 3 main categories - dermal, vascular and ground tissues. There are three types of plant tissues - meristematic, permanent and complex permanent tissues. Meristematic tissues are found in apical, lateral and intercalary meristems and allow for plant growth. Permanent tissues such as parenchyma, collenchyma and sclerenchyma do not divide. Complex permanent tissues include xylem and phloem which transport water and nutrients. Plant tissues have many economic uses including food, furniture, clothing fibers, horticulture, and conservation of endangered species.
This document describes and compares the internal structures of dorsiventral and isobilateral leaves. A dorsiventral leaf, characteristic of dicots, has distinct upper and lower epidermis, with palisade and spongy mesophyll layers and stomata only on the lower epidermis. An isobilateral leaf, found in monocots, has a uniform internal structure with stomata on both surfaces and spherical mesophyll cells without differentiation. Key differences are summarized in a table contrasting features such as venation patterns and distribution and types of epidermal and mesophyll cells between the two leaf types.
Cyanobacteria, also known as blue-green algae, are photosynthetic prokaryotes that reproduce through binary fission and have specialized structures like hormogones, akinetes, and heterocysts that aid in reproduction and survival. They have a multilayer cell wall and lack organelles but contain pigments like chlorophyll and phycobilins within thylakoid membranes to perform photosynthesis. Cyanobacteria inhabit various environments like freshwater, marine water, soil and hot springs.
The document discusses plant growth and development. It defines growth as a permanent increase in plant mass through cell division and enlargement. Growth occurs in phases including formation, elongation, and maturation. Development includes differentiation, where cells take on specialized structures and functions. The document also discusses leaf growth, flowering, pollination, fertilization, seed formation, and fruit development. Environmental factors and phytohormones influence growth and developmental processes.
This document discusses maceration and smear techniques. Maceration involves separating plant or animal tissues into individual cells or groups by dissolving the middle lamella. Three common maceration methods are described: Schultze's method using nitric acid and potassium chlorate, Jeffrey's method using boiling and chromic acid, and Harlow's method using chlorine water. Smears are used to study internal cell details like nuclei and are prepared by smearing material on a slide. Two staining methods for smears are acetocarmine, which stains and fixes in one step, and Feulgen's method using basic fuchsin.
MICROTECHNIQUE Killing and fixation moduleAbdulsalm
This document provides instructions for preparing biological specimens for light microscopy. It discusses the key steps of sample collection, killing and fixation, dehydration, clearing, paraffin embedding, microtomy, staining, and observation. Specific fixation fluids, dehydration reagents, clearing agents, and staining methods are described. The goal is to preserve specimens while modifying properties like refractive index to allow examination under a light microscope.
This document provides an overview of cryopreservation, which involves preserving biological material such as cells, tissues, organs, and embryos at ultra-low temperatures, typically in liquid nitrogen. It discusses the history, principles, mechanisms, and applications of cryopreservation. Key aspects covered include the use of cryoprotectants to prevent freezing damage to cells, various freezing and thawing methods, long-term storage in liquid nitrogen, and viability testing after thawing to regenerate plants or animals from preserved material. Cryopreservation has important applications in biobanking, conservation of endangered species, and preservation of disease-free agricultural crops.
Auxins are a specific group of plant hormones that play an important role in plant growth and development. Some key points about auxins include:
- They were the first plant hormone discovered. Common auxins include indole-3-acetic acid (IAA).
- Auxins are synthesized in root tips, young leaves, and apical buds, and are involved in processes like stem elongation, root initiation, apical dominance, and fruit development.
- Their discovery was made through experiments by Charles Darwin, Peter Boysen-Jensen, and Frits Went in the late 19th/early 20th centuries which demonstrated that a signal substance moved through plants and caused differential growth.
Tissue culture involves growing plant and animal cells, tissues, or organs artificially in a sterile environment with a nutrient medium. It allows for cloning and mass propagation of plants and animals. Tissue culture requires selecting appropriate explant tissue, developing a sterile culture medium with growth regulators, and regularly subculturing and multiplying the cells. It has many applications like rapidly propagating desirable crops and endangered species, providing pathogen-free plants, selecting for improved varieties, and studying cell processes. Animal tissue culture differs in requiring serum, having limited cell divisions, and posing some biohazard risks. Its uses include growing viruses, producing monoclonal antibodies, and facilitating genetic studies.
Plant tissue culture is a collection of techniques used to grow plant cells, tissues or organs under sterile conditions. It allows for the mass production of clones of plants with desirable traits. The key aspects of plant tissue culture are maintaining sterile conditions on a nutrient medium, and providing proper aeration. Common types of plant tissue culture include callus culture, single cell culture, root tip culture, shoot tip culture, and anther culture. Plant tissue culture has many applications for plant conservation, breeding, and production of secondary metabolites.
The fungal cell wall is composed of various components that give shape, strength, and protection to fungi. The main components are chitin, glucan, and proteins. Chitin is a nitrogenous polysaccharide made of N-acetylglucosamine that forms microfibrils providing structure. Glucan is a polysaccharide of glucose monomers linked by beta glycosidic bonds that is a major component of fungal cell walls. Glycoproteins and proteins are also embedded in the cell wall matrix and contain carbohydrates. Together these components form the primary wall and secondary wall layers that define fungal cells.
Fungi have several distinguishing morphological features:
1. They have cell walls containing chitin and lack peptidoglycan.
2. They can exist in both unicellular and multicellular forms, dividing asexually or sexually.
3. They are classified based on their structures - yeasts are unicellular, molds form branching hyphae and mycelium, and dimorphic fungi switch between yeast and mold forms based on temperature.
The isolation, culture and fusion of protoplasts is a fascinating field in plant research. Protoplast isolation and their cultures provide millions of single cells (comparable to microbial cells) for a variety of studies.
Vital staining involves using non-toxic dyes on living organisms and cellular structures. Common vital stains include neutral red, methylene blue, and Janus green B. Vital stains can be used to differentiate between living and dead cells, stain specific organelles like mitochondria, and study pathological cell changes without harming living tissues. Vital stains are classified based on their chemical properties and degree of dissociation into categories like basic, acidic, atmospheric and electro neutral. Techniques for vital staining include progressive, regressive, counterstaining, and double staining using combinations of contrasting dyes.
basic principles and protocol in plant tissue cultureAlia Najiha
Plant tissue culture involves growing plant cells, tissues, or organs in an artificial nutrient medium under sterile conditions. It allows for the rapid production of exact genetic copies of plants and is used to regenerate whole plants from genetically modified plant cells. The key requirements for successful tissue culture are appropriate explant tissue, a suitable growth medium, aseptic conditions to prevent microbial contamination, growth regulators, and frequent subculturing to provide nutrients and remove waste. The basic steps are selection of explant tissue from a healthy mother plant, sterilization of the tissue, establishment of the explant in a culture medium, multiplication through callus formation and shoot development, and root formation through manipulation of growth regulators. Benefits include providing pathogen-free plants
The document discusses the structure and functions of plant stems. It begins by defining stems and describing their main functions: support, conduction, and production of new living tissue. It then discusses the external morphology of stems, including nodes, internodes, buds, and other features. The internal structure is also examined, focusing on the shoot apex organization and primary meristems that give rise to primary tissues like epidermis, ground tissue, and vascular tissues. Primary growth and tissue development in dicot and monocot stems is contrasted. Finally, the document covers secondary growth, which occurs in woody stems through the activity of the vascular cambium and cork cambium.
Cryopreservation is the process of preserving living cells and tissues by cooling them to low sub-zero temperatures. This stops any chemical or enzymatic activity in the cells that could cause damage. Traditional cryopreservation relies on coating materials in cryoprotectants like glycerol or DMSO to prevent ice formation during freezing. Cryopreservation has applications for preserving reproductive cells, embryos, and tissues and is also used in the unproven field of cryonics which seeks to preserve entire human bodies for possible future revival.
This document provides instructions for sterilizing plant explants for tissue culture. It discusses that culture media supports microbial growth which can kill plant tissues, so complete sterilization is essential. The key steps outlined are: washing explants with detergent and water; submerging in disinfectants like ethanol or mercury chloride; rinsing with sterile water; and sealing cultures with sterile cotton plugs. A two-step sterilization using ethanol prior to another disinfectant can improve sterility. Proper sterilization will increase the success rate of tissue culture by reducing contamination and allowing explant growth without microbes.
Gibberellic Acid or Gibberellin Hormonesvidan biology
Gibberellins (GAs) are plant hormones that regulate various developmental processes. They are tetracyclic diterpenoid acids synthesized via the terpenoid pathway in plastids and then modified in the endoplasmic reticulum and cytosol. Bioactive GAs include GA1, GA3, GA4, and GA7 and contain a carboxyl group at C-7. GAs are synthesized in shoots and roots and translocated via phloem and xylem, respectively. DELLA proteins repress growth but are degraded by the 26S proteasome in response to GA binding to the GID1 receptor, which forms a complex targeting DELLA for ubiquitination and degradation. This releases repression
Cytokinins are a class of plant growth hormones that promote cell division. They were first discovered in 1955 from degraded herring sperm DNA. There are two main types: adenine-type cytokinins like kinetin and zeatin, and phenylurea-type cytokinins. Cytokinins play many important physiological roles in plants including cell division, cell elongation, differentiation, delaying senescence, and influencing flowering. They are widely used in tissue culture and to increase the shelf life of cut flowers and vegetables.
Plant tissues can be divided into 3 main categories - dermal, vascular and ground tissues. There are three types of plant tissues - meristematic, permanent and complex permanent tissues. Meristematic tissues are found in apical, lateral and intercalary meristems and allow for plant growth. Permanent tissues such as parenchyma, collenchyma and sclerenchyma do not divide. Complex permanent tissues include xylem and phloem which transport water and nutrients. Plant tissues have many economic uses including food, furniture, clothing fibers, horticulture, and conservation of endangered species.
This document describes and compares the internal structures of dorsiventral and isobilateral leaves. A dorsiventral leaf, characteristic of dicots, has distinct upper and lower epidermis, with palisade and spongy mesophyll layers and stomata only on the lower epidermis. An isobilateral leaf, found in monocots, has a uniform internal structure with stomata on both surfaces and spherical mesophyll cells without differentiation. Key differences are summarized in a table contrasting features such as venation patterns and distribution and types of epidermal and mesophyll cells between the two leaf types.
* Treponema pallidum causes syphilis.
* A highly aneuploid tumor indicates genomic instability which is associated with increased aggressiveness and poor prognosis.
* Amyloid protein showing apple green birefringence under polarized light microscopy has clinical significance as it indicates amyloidosis.
* The signs ±, ± ± indicate the type of birefringence shown by substances under polarized light microscopy. ± indicates negative birefringence shown by uric acid while ± ± indicates positive birefringence shown by calcium pyrophosphate.
* Osmium tetroxide reacts with phospholipids in cell membranes, making them electron dense and visible under electron microscopy. This helps in better structural visualization of cells
The document discusses the key steps in histological techniques which are used to prepare tissue samples for microscopic examination. The techniques include fixation, dehydration, clearing, embedding, sectioning, and staining of tissues. Fixation preserves the tissue structure using chemical fixatives. The tissue then undergoes dehydration, clearing and infiltration with paraffin wax before being embedded in wax blocks and sectioned for microscopic analysis. Staining helps to visualize different structures within the tissue sample under the microscope.
This document discusses specialty food and beverage insurance provided by Personal Coverage, Inc. It covers commercial general liability insurance, including bodily injury and property damage liability limits. It also addresses policy features like deductibles, additional insureds, territory coverage, and whether defense costs are included within or outside the liability limits. The document provides an overview of important questions to consider about liability insurance, such as whether commercial general liability or product liability is needed, ensuring limits meet contractual requirements, occurrence vs. claims-made policies, and how to cancel a policy if no longer required. It introduces Tom Wallace as the executive VP who can be contacted for more information.
This document discusses resiliency and provides strategies for building resilience. It defines resilience as "the ability to withstand, recover and/or grow in the face of stressors and changing demands." The document notes that resilience allows people to bounce back from difficulties and leads to good outcomes. It encourages focusing on the future rather than dwelling on the past and learning from experiences to move forward. The conclusion emphasizes that resilience is a skill that can be improved and that everyone is capable of coping with challenges and rebounding from setbacks.
Best professional indemnity insurance quotekelly657
This website provides information about professional indemnity insurance and obtaining quotes. It explains that professional indemnity insurance protects businesses and individuals against claims for compensation arising from negligent acts, errors or omissions in their work. The site allows users to enter some basic details to receive customized quotes for professional indemnity insurance from various insurers.
Pre-Con Ed: Covering Your "Assets" - Don't get Caught with Your [Software] Pa...CA Technologies
The document discusses using connectors in CA IT Asset Manager to collect software asset management data from multiple sources. It describes the different types of data sources, including primary sources that create device records and enrichment sources that add additional data to existing records. It emphasizes finding the right data sources to collect the necessary metrics for effective software license compliance calculations. Standard connectors are available for common inventory and license data sources, while custom connectors can be created for other sources. The Data Coordination Service acts as an out-of-the-box enrichment source to consolidate asset data.
Cost and Time savings through ValGenesis -Validation Software SolutionValGenesis
As a pioneer in the development of automated validation software solutions, ValGenesis, Inc. leads the way with a unique paperless validation process that tracks the validation status of any GxP system in real time. With years of development and refinement in place,
ValGenesis is the first to offer users real-time paperless validation via electronic execution thereby minimizing ineffectiveness and maximizing efficiency.
ValGenesis automates and manages the validation life cycle and provides real time validation status of any system corporate wide.
The document summarizes the process of tissue processing which involves fixing, dehydrating, clearing, infiltrating with wax, embedding, sectioning, staining, and mounting tissue samples in order to examine them microscopically. Key steps include fixation to prevent decay, dehydration using alcohol to remove water, clearing with xylene to remove alcohol, infiltration and embedding in paraffin wax, sectioning thin slices with a microtome, staining typically with hematoxylin and eosin for examination, and mounting on slides. The goal is to prepare tissue for microscopic analysis while maintaining structure.
PREPARATION OF HISTOLOGICAL SPECIMENS.pptxAnthonyMatu1
This document provides information on the processes involved in preparing histological specimens, including tissue fixation, processing, sectioning, and staining. It describes the objectives of tissue fixation as preventing autolysis and bacterial attack while maintaining tissue structure. The main types of fixatives and factors affecting fixation are outlined. Tissue processing involves dehydration using graded alcohols, clearing with agents like xylene, and embedding in paraffin wax. Sections are cut with a microtome and stained, with hematoxylin and eosin being a common staining method that colors nuclei blue and cytoplasm pink. Other specialized staining techniques are also mentioned.
Microtomy, or the preparation of tissue slides, is the foremost technique used in histological studies. This presentation is a brief overview of the technique and the steps involved.
HISTOTECHNIQUES MICROTOMY.ppt · version 1.pdfmahrukhmughal27
1. Histotechniques involve processing tissues through fixation, dehydration, clearing, embedding, section cutting, and staining to enable pathological examination under a microscope.
2. Tissues are first fixed in chemicals like formaldehyde to preserve their structure, then dehydrated with graded alcohols, cleared with solvents, and embedded in paraffin wax for section cutting with a microtome.
3. The microtome uses a knife to cut extremely thin sections of the wax-embedded tissue, which are then floated in water, mounted on slides, and stained for microscopic examination.
Histological techniques involve fixing, processing, sectioning, and staining tissue samples to examine their microscopic structure. Fixation preserves tissues from degradation. Processing dehydrates tissues and embeds them in paraffin wax or other materials to allow thin sectioning. A microtome is used to cut thin sections for staining with dyes like hematoxylin and eosin, which impart color to different tissue components to reveal their structure under a microscope. These techniques prepare tissues for microscopic analysis while minimizing artifacts from handling.
Histological techniques involve preparing tissue for microscopic examination through a series of procedures to retain the tissue's microscopic anatomy and allow for good staining. This involves fixing, processing, embedding, sectioning, and staining the tissue. The key steps are fixation to prevent deterioration, dehydration and clearing to infiltrate the tissue with wax, sectioning thin slices, and staining, most commonly with hematoxylin and eosin, to highlight structures. The goal is to represent the tissue's structure as closely as possible to its in vivo state.
This document provides an introduction to histopathology and the process of tissue fixation. It defines histopathology as the microscopic examination of tissue to study disease. The key steps in tissue fixation are described as collection and transport of biopsy specimens, followed by processing including fixation, dehydration, clearing, embedding, cutting and staining of tissues. Critical factors that ensure proper fixation include freshness of tissues, penetration of fixatives, duration of fixation, and choice of fixative based on the tissue and intended analysis. Formalin is the most commonly used fixative but others including glutaraldehyde and osmium tetroxide are also described.
This document outlines the histotechnique process which tissues undergo before microscopic examination. Key steps include: fixation to preserve tissue structure; processing involving dehydration, clearing, and impregnation to allow sectioning; embedding tissues in paraffin blocks for microtomy; sectioning samples and staining, typically with hematoxylin and eosin, for visualization under the microscope. Finally, samples are mounted on slides and labeled for storage and pathological examination.
Microtomy, or the preparation of tissue slides, is the foremost technique used in histological studies. This is a brief overview of the technique and the steps involved.
Histopathology techniques are used to demonstrate minute structural alterations in tissues caused by disease. Key techniques include fixing tissues in formalin to preserve structure, processing tissues through dehydration, clearing and infiltration steps, embedding in paraffin wax, sectioning with a microtome, staining, and mounting slides. Histopathology allows diagnosis of diseases through microscopic examination of tissue structures and any pathological changes present. It is a crucial technique when other testing may not be possible or provides definitive confirmation of diseases.
This document provides an overview of histology, the study of tissues. It discusses that tissues are made up of cells and an extracellular matrix that interact closely. The key steps in preparing tissues for microscopic examination are fixation, dehydration, clearing, embedding, sectioning, and staining. Fixation preserves tissues using chemicals like formaldehyde. Sections are cut very thin using a microtome and mounted on slides for staining with dyes like hematoxylin and eosin or periodic acid-Schiff to visualize different tissue components under the microscope. The interactions between cells and extracellular matrix allow tissues to form and carry out specialized functions in organs.
This document provides an overview of histopathology and the process of tissue fixation and preparation for microscopic examination. It defines histopathology and describes the various steps tissues undergo, including fixation, to arrive at a diagnosis. It discusses the aims and principles of fixation, including preventing autolysis and putrefaction. Various fixatives are described, including formalin, alcohol, mercuric chloride and picric acid. The document also covers tissue processing, including sectioning and staining, as well as the roles and responsibilities of laboratory technicians in specimen handling.
This document provides an introduction to histopathology and the process of tissue sample preparation and analysis. It defines histopathology and describes the steps tissues undergo, including fixation, processing, sectioning and staining. It discusses the types of samples obtained, including biopsies and autopsy tissues. It explains the responsibilities of laboratory technicians in handling, labeling and storing samples. Finally, it provides details on fixation, including the goals, common fixatives used, and how to properly prepare tissue samples for fixation.
Immunohistochemistry (IHC) is a method that combines biochemical, histological and immunological techniques into a simple but powerful assay for protein detection. IHC provides valuable information as it visualizes the distribution and localization of specific cellular components within cells and in proper tissue context.
1. The document discusses various cytological techniques used in the study and manipulation of cells including microtomy, microscopy, staining, and centrifugation.
2. Microtomy involves sectioning tissues into thin slices using a microtome for microscopic observation while staining allows cellular structures to be visualized through selective uptake of dyes.
3. Centrifugation separates particles according to properties like density, shape and size through application of centrifugal force, and is used to isolate cellular components through differential pelleting or density gradient centrifugation.
1. The document discusses various cytological techniques used in the study and manipulation of cells including microtomy, microscopy, staining, and centrifugation.
2. Microtomy involves sectioning tissues into thin slices using a microtome for microscopic examination after fixation, dehydration, embedding, and staining.
3. Centrifugation separates particles in a solution based on properties like density, shape, and size through application of centrifugal force. Types include differential pelleting and density gradient centrifugation.
Practical Histopathology and cytopathology
Histopathological examination is used to provide diagnostic information that is important for timely diagnosis of disease to determine treatment plan. Fresh tissue is extremely fragile & subject to autolysis.
This document discusses various techniques used in histopathology. It defines histology and histopathology and describes fixation techniques like formalin fixation which preserves tissues. The steps of tissue processing like dehydration, clearing, impregnation and embedding in paraffin wax are explained. Microtomy or section cutting and staining techniques including hematoxylin and eosin staining are also summarized. The overall techniques discussed allow examination of tissues at a microscopic level for abnormalities.
This document discusses various techniques used in histopathology. It defines histology and histopathology and describes fixation techniques like formalin fixation which preserves tissues. The steps of tissue processing like dehydration, clearing, impregnation and embedding in paraffin wax are explained. Microtomy or section cutting and staining techniques including hematoxylin and eosin staining are also summarized. The overall techniques discussed allow examination of tissues at a microscopic level for abnormalities.
This document provides an overview of histology and microscopy. It discusses the four basic tissue types studied in histology, as well as techniques for tissue collection, processing, fixation, embedding and section cutting to prepare samples for microscopic examination. It also describes the principles of light microscopy and electron microscopy, and different types of microscopes used including bright field, dark field, phase contrast and fluorescence microscopes. The importance of histology for understanding the relationship between tissue structure and function is emphasized.
5. Biopsy
Removal of a sample of living tissue for
microscopic examination.
Used to diagnose cancers, infections, etc
6. How to get tissues for study
Steps in tissue preparation
Fresh tissues from the body
1. fixation
Formalin ( 10% formaldehyde)
Osmium tetroxide for EM
Mechanism - Forms cross links with proteins (Lysine)
2. Embedding – gives support for tissue slicing
Paraffin or plastic resin
3. Washing & dehydration (dehydration by graded alcohols in ascending
order)
4. clearing – to remove paraffin & alcohol
By xylol or tulol
5. block making
How to generate histology slides?
7. 6. section cutting – 5-10μ thick sections with microtome
7. mounting – on glass slide ( adhesive – albumin)
8. clearing – xylol / tulol
9. rehydrate – alcohols in descending order
Staining
nuclear stain – Hematoxylin ( basic stain & water soluble)
counter stain – Eosin ( less water soluble but soluble in alcohol)
– dehydrate in ascending order
10. Clearing – xylol / tulol
11.Mounting medium – cover glass
How to generate histology slides?
8. TISSUE FIXATION
Fixation is a complex series of chemical events that differ for the
different groups of substance found in tissues.
The aim of fixation:
1- To prevent autolysis and bacterial attack.
2- To fix the tissues so they will not change their volume and shape
during processing.
3- To prepare tissue and leave it in a condition which allow clear
staining of sections.
4- To leave tissue as close as their living state as possible, and no
small molecules should be lost.
Fixation is coming by reaction between the fixative and protein
which form a gel, so keeping every thing as their in vivo relation to
each other.
9. Factors affect fixation:
- PH.
- Temperature.
- Penetration of fixative.
- Volume of tissue.
According to previous factors we can determine the concentration of
fixative and fixation time.
Types of fixative:
Acetic acid, Formaldehyde, Ethanol, Glutaraldehyde, Methanol and
Picric acid.
10. TISSUE PROCESSING
the aim of tissue processing is to embed the tissue in a solid
medium firm enough to support the tissue and give it sufficient
rigidity to enable thin sections to be cut , and yet soft enough not to
damage the knife or tissue.
Stages of processing:
1- Dehydration.
2- Clearing.
3- Embedding.
11. Dehydration
to remove fixative and water from the tissue and replace
them with dehydrating fluid.
There are a variety of compounds many of which are
alcohols. several are hydrophilic so attract water from
tissue.
To minimize tissue distortion from diffusion currents,
delicate specimens are dehydrated in a graded ethanol
series from water through 10%-20%-50%-95%-100%
ethanol.
In the paraffin wax method, following any necessary post
fixation treatment, dehydration from aqueous fixatives is
usually initiated in 60%-70% ethanol, progressing
through 90%-95% ethanol, then two or three changes of
absolute ethanol before proceeding to the clearing stage.
12. Types of dehydrating agents:
Ethanol, Methanol, Acetone.
Duration of dehydration should be kept to the minimum consistent
with the tissues being processed. Tissue blocks 1 mm thick should
receive up to 30 minutes in each alcohol, blocks 5 mm thick require
up to 90 minutes or longer in each change. Tissues may be held and
stored indefinitely in 70% ethanol without harm
13. Clearing
replacing the dehydrating fluid with a fluid that is totally miscible with
both the dehydrating fluid and the embedding medium.
Choice of a clearing agent depends upon the
following:
- The type of tissues to be processed, and the type of processing to
be undertaken.
- The processor system to be used.
- Intended processing conditions such as temperature, vacuum and
pressure.
- Safety factors.
- Cost and convenience.
- Speedy removal of dehydrating agent .
- Ease of removal by molten paraffin wax .
- Minimal tissue damage .
15. Embedding
is the process by which tissues are surrounded by a medium such
as agar, gelatin, or wax which when solidified will provide sufficient
external support during sectioning.
Paraffin wax
properties :
Paraffin wax is a polycrystalline mixture of solid hydrocarbons
produced during the refining of coal and mineral oils. It is about two
thirds the density and slightly more elastic than dried protein.
Paraffin wax is traditionally marketed by its melting points which
range from 39°C to 68°C.
The properties of paraffin wax are improved for histological
purposes by the inclusion of substances added alone or in
combination to the wax:
- improve ribboning.
- increase hardness.
- decrease melting point
- improve adhesion between specimen and wax
16. Precaution while embedding in wax
The wax is clear of clearing agent.
No dust particles must be present.
Immediately after tissue embedding, the wax must be rapidly cooled
to reduce the wax crystal size.
17. Tissue processing
Embedding moulds:
(A) paper boat;
(B) metal bot mould;
(C) Dimmock embedding mould;
(D) Peel-a-way disposable mould;
(E) base mould used with embedding
ring ( F) or cassette bases (G)
18. General Embedding Procedure
1- Open the tissue cassette, check against worksheet entry to ensure the
correct number of tissue pieces are present.
2- Select the mould, there should be sufficient room for the tissue with
allowance for at least a 2 mm surrounding margin of wax.
3- Fill the mould with paraffin wax.
4 Using warm forceps select the tissue, taking care that it does not cool in
the air; at the same time.
5- Chill the mould on the cold plate, orienting the tissue and firming it into
the wax with warmed forceps. This ensures that the correct orientation is
maintained and the tissue surface to be sectioned is kept flat.
6- Insert the identifying label or place the labeled embedding ring or
cassette base onto the mould.
7- Cool the block on the cold plate, or carefully submerge it under water
when a thin skin has formed over the wax surface.
8- Remove the block from the mould.
9- Cross check block, label and worksheet.
19.
20. ORIENTATION OF TISSUE IN THE BLOCK
Correct orientation of tissue in a mould is the most important step in
embedding. Incorrect placement of tissues may result in
diagnostically important tissue elements being missed or damaged
during microtomy.
elongate tissues are placed diagonally across the block
tubular and walled specimens such as vas deferens, cysts and
gastrointestinal tissues are embedded so as to provide transverse
sections showing all tissue layers
tissues with an epithelial surface such as skin, are embedded to
provide sections in a plane at right angles to the surface (hairy or
keratinised epithelia are oriented to face the knife diagonally)
multiple tissue pieces are aligned across the long axis of the mould,
and not placed at random
23. A microtome is a mechanical instrument
used to cut biological specimens into very
thin segments for microscopic
examination. Most microtomes use a steel
blade and are used to prepare sections of
animal or plant tissues for histology. The
most common applications of microtomes
are
25. 1- Traditional histological technique:
tissues are hardened by replacing water with paraffin. The tissue is
then cut in the microtome at thicknesses varying from 2 to 25
micrometers thick. From there the tissue can be mounted on a
microscope slide, stained and examined using a light microscope
26. 2- Cryosection:
water-rich tissues are hardened by freezing and cut frozen;
sections are stained and examined with a light microscope.
This technique is much faster than traditional histology (5
minutes vs. 16 hours) and are used in operations to achieve a
quick diagnosis. Cryosections can also be used in
immunohistochemistry as freezing tissue does not alter or
mask its chemical composition as much as preserving it with a
fixative.
28. H & E is a charge-based, general purpose stain. Hematoxylin
stains acidic molecules shades of blue. Eosin stains basic
materials shades of red, pink and orange. H & E stains are
universally used for routine histological examination of tissue
sections.
Hematoxylin and Eosin (H & E)
29. Fixation
Any well fixed tissue.
Staining Procedure
1- Deparaffinize and hydrate to water
2- If sections are Zenker-fixed, remove the mercuric chloride crystals
with iodine and clear with sodium thiosulphate (hypo)
3- Mayer's hematoxylin for 15 minutes
4- Wash in running tap water for 20 minutes
5- Counterstain with eosin from 15 seconds to 2 minutes depending
on the age of the eosin, and the depth of the counterstain desired.
For even staining results dip slides several times before allowing
them to set in the eosin for the desired time
6- Dehydrate in 95% and absolute alcohols, two changes of 2
minutes each or until excess eosin is removed. Check under
microscope
7- Clear in xylene, two changes of 2 minutes each
8- Mount in Permount or Histoclad
Results
Nuclei - blue - with some metachromasia
Cytoplasm - various shades of pink-identifying different tissue
components
36. Staining – routine stain – H&E
Some structures are seen/ preserved (large molecules like
nucleoproteins, cytoskeleton proteins, ECM proteins- collagen,
membrane proteins)
some are not seen/lost (small molecules -t-RNA, large molecules like
glycogen & Proteioglycans are dissolved, )during the fixation/staining
process
Special fixatives to retain membrane ( phospholipids)
Permanganate & osmium – for EM
For Elastic fibers – Orcein/ Resorcin – Fuscin
For reticular fibers – Silver impregnation
Histochemistry & Cytochemistry
Specific binding of dye with particular molecule
Fluorescent dye labeled antibody to cell component
Enzyme activity
Autoradiography – radio isotopes tagged with precursors of a
molecule molecule incorporated into cell/ tissue before fixation
Special situations
40. Gomori trichrome stain
Gomori's one-step trichrome is
a staining procedure that
combines the plasma stain
(chromotrope 2R) and
connective fiber stain (fast
green FCF) in a
phosphotungstic acid solution
to which glacial acetic acid has
been added.
http://freepages.genealogy.rootsweb.ancestry.com/~gomery/gomorigeo.html
41. Trichrome stain (Generally Masson’s) –
To delineate cells from surrounding connective Tissue
www-bioc.rice.edu/bios576/immuno/Trichrome.jpg
42. Special stain
PAS positive substances Carbohydrate
(glycogen) or carbohydrate rich molecules,
Basement membrane, reticular fibers
Periodic acid cleaves bond between carbon atoms
form aldehyde group
Aldehyde binds with Schiff to produce magenta or
pink color
PAS =Periodic Acid Schiff
45. Acid hydrolyses or cleaves proteins from
deoxyribose of DNA leads to opening of
sugar group & formation of aldehyde
Schiff binds and gives magenta color to
aldehyde
Can be useful to quantify amount of DNA ( by
using spectrophotmetry of Feulgen stained
tissue)
Feulgen stain for Nuclear Proteins
Why RNA cannot be stained by Feulgen?
46. For the confirmation of specific substances
Pretreatment of sections with specific
enzymes
Diastase/amylase for glycogen
DNA ase for DNA
Enzymatic digestion
47. Figure 1—17.
Photomicrograph of a rat
kidney section treated by
the Gomori method to
demonstrate the enzyme
alkaline phosphatase. The
sites where this enzyme is
present (cell surface) stain
intensely with black
(arrows). Medium
magnification.
50. Figure 1—16. Photomicrograph of
a bone section treated with a
histochemical technique to
demonstrate calcium ions. The
dark precipitate indicates the
presence of calcium phosphate in
calcified bone and cartilage.
Noncalcified cartilage tissue
(stained in pink) is in the upper
portion of the figure. Medium
magnification.
51. Antibody ( Immunoglobulin) conjugated with
fluorescent dye( most common is Fluorescein)
+ Antigen ( foreign protein)
Fluorescein absorbs UV light and emits
green fluorescence can be seen under
Fluorescent microscope (IF- Immuno
Fluorescence)
Example :- actin (Antigen) of Rat infected
to Rabbit blood of Rabbit ( have poly -
clonal antibodies for Rat’s actin/ anti rat actin
antibodies) bind with Fluorescent dye
Immuno Histo Chemistry (IHC)
52. Specific antigen
(actin of rat)
Monoclonal Antibodies
Multiple Myeloma pts.
Monoclonal B ells
Hybridoma cells
↓
Single specific type of antibodies (Monoclonal)
( against Actin)
B lymphocytes of
Immunized rabbit
↓
53. Diagnosis of tumors(tumor markers) &
Infections( HIV, Infectious Mononucleosis)
Classify sub – types (B -cell and T- cell
lymphomas)
Treatment – Anti-TNF-α antibodies in
inflammatory disorders
Clinical Significance of Monoclonal
Antibodies
57. Figure 1—26. Photomicrograph
of a section of small intestine in
which an antibody against the
enzyme lysozyme was applied
to demonstrate lysosomes in
macrophages and Paneth cells.
The brown color results from
the reaction done to show
peroxidase, which was linked to
the secondary antibody. Nuclei
counterstained with
hematoxylin. Medium
magnification.
58. Figure 1—28. Electron micrograph showing a section of a pancreatic acinar cell
that was incubated with anti-amylase antibody and stained by protein A coupled
with gold particles. Protein A has high affinity toward antibody molecules. The
gold particles appear as very small black dots over the mature secretory granules
and the forming granules in the Golgi complex. (Courtesy of M Bendayan.)
59. Localization of enzymatic activity in tissues
Best fixation – mild aldehyde ( formalin)
Basis – localized reaction production of
enzyme activity
Used for acid & alkaline phosphatase, ATP
ases
AB (substrate) + T (trap) AT (
reaction product) + B (Hydrolyzed
component of substrate)
Enzyme Histochemistry
enzyme
60. Other Methods
Hybridization: for localizing
mRNA/DNA (NA)
In Situ Hybridization: Binding (
Probe + NA) in cell/tissue
FISH: If Fluorochrome is used
in Hybridization technique
Autoradiography: by tagging
the precursor molecules (Amino
acids) followed by synthesis of
large molecules (NA) localize
the particular tagged molecule
63. 3- Electron microscopy:
after embedding tissues in epoxy resin, a microtome equipped with
a glass or diamond knife is used to cut very thin sections (typically
60 to 100 nanometers). Sections are stained and examined with a
transmission electron microscope. This instrument is often called an
ultramicrotome.