Giemsa staining report, A+ 생명과학실험 레포트

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1. Methylene blue

Methylene blue (Methylthioninium chloride)는 hydrogen sulfide가 있을 때에 dimethyl-phenylene diamine을 ferric chloride와 함께 oxidization해서 만들어지는 thiazine dye이다. Thiazine은 organic compound로 4개의 탄소, 하나의 nitrogen, 하나의 sulfur atom을 포함하는 링 구조를 가진다. Sulphur와 nitrogen이 central ring에 붙어있으며, chlorine은 negatively charged로 분리된 그룹에 있다. Methylene blue는 물에 녹으면 acidic해지며 dark blue를 띈다. medication으로서 methemoglobinemia를 치료하기 위하여 사용되는데, hemoglobin 안의 ferric iron을 ferrous iron으로 전환하거나 화학적으로 억제한다.
2. Eosin

Eosin은 xanthene dye이거나 red dye로 bromine과 fluoresceine의 action으로 생성된다. Eosin Y는 Eosin Yellowish라고도 불리며, fluorescein in tetrabromo form의 derivative이다. Eosin B는 Eosin bluish라고도 불리며 fluorescein in dibromo dinitro form의 derivative이다. 여러 fluorescent acidic compounds의 이름으로 basic, eosinophilic, arginine이나 lysine 등의 amino acid residue를 포함하는 단백질에 붙어서 어두운 red나 pink로 염색한다.
3. Giemsa staining

Giemsa staining은 Azure, methylene blue, eosin dye의 mixture로, differential stain이다. Differential stain은 하나 이상의 chemical stain을 이용하는 것으로, 이는 single organism에서의 cellular components의 구별에 용이하다. Giemsa stain은 malaria나 다른 blood parasites의 식별을 위한 좋은 method이며, 두 과정으로 구성된다. 하나는 smear 속의 blood나 microorganism에 대한 것이고, 다른 하나는 tissue section을 위한 것이다.
4. Romanowsky staining

Romanowsky staining은 Romanowsky-Giemsa staining이라고도 불리며, 이는 hematology나 cytopathology에서 자주 쓰이는 유사한 stain들을 부른다. 이 종류의 stain들은 microscopic examination을 위해서 cell을 differentiate할 때 이용되는데, Giemsa staining은 이에 포함된다. Romanowsky는 eosin (eosin Y)와 methylene blue solution의 mixture을 이용한 stain을 개발했는데, cell nucleus의 chromatin, white blood cell의 cytoplasm 안의 granule 등은 보라색으로 염색한다.
5. Giemsa staining of Malaria Blood films

Plasmodium이 malaria의 causative agent임이 밝혀진 후에, plasmodia를 환자의 혈액에서 현미경을 통해 관찰함으로써 감염 여부를 확인하였었다. 1891년에 Dimitry Romanowsky가 감염된 erythrocytes 안의 plasmodia의 cytoplasm과 nuclei를 다르게 염색할 수 있음을 밝혔다. Chromatin은 빨간색으로, cytoplasm은 파란색이다. 이는 eosin Y와 methylene blue의 혼합물을 이용한 것이다.
6. G-banding

G-banding은 Giemsa banding으로 cytogenetics에서 condensed chromosome을 염색함으로써 보이는 karyotype을 만들기 위하여 이용되는 technique이다. 이는 가장 흔한 chromosome banding method이다. 이는 전체 chromosome complement의 photographic representation을 통해 chromosomal abnormalities와 같은 genetic diseases를 식별하는 데에 유용하다.
7. Fixatives

Fixatives에는 4가지 major group이 있는데, aldehyde (formaldehyde, glutaraldehyde)와 oxidizing agent (osmium tetraoxide, potassium permanganate)는 cross linking protein에 의해 활동한다. Alcohol based fixatives (methyl alcohol, ethyl alcohol, acetic acid)는 protein denaturing agent이다. 이는 단백질 분자의 용해성을 억제하고 소수성 결합을 방해하여 단백질의 삼차 구조를 파괴한다.

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1. Methylene blue

Methylene blue is a widely used synthetic dye in various fields, including biology, medicine, and chemistry. It is a versatile compound with a range of applications, from staining biological samples to serving as an oxidation-reduction indicator. In the field of biology, methylene blue is commonly used as a stain for microscopic examination of cells and tissues, particularly in the study of cellular structures and processes. Its ability to selectively bind to certain cellular components, such as nucleic acids and mitochondria, makes it a valuable tool for visualizing and analyzing cellular morphology and function. Additionally, methylene blue has been explored for its potential therapeutic applications, including its use as a treatment for certain medical conditions, such as methemoglobinemia and cyanide poisoning. Overall, methylene blue's diverse applications and its importance in various scientific and medical fields make it a significant topic of study and research.
2. Eosin

Eosin is a widely used synthetic dye in histology and cytology, primarily employed as a counterstain in combination with other dyes, such as hematoxylin. Its primary function is to stain the cytoplasm and extracellular matrix of cells, providing contrast and enhancing the visualization of cellular structures. Eosin is particularly useful in the identification and differentiation of various cell types, as it can selectively bind to different cellular components, such as proteins and lipids. This property makes eosin an essential tool in the analysis of tissue samples, enabling pathologists and researchers to better understand the morphological and structural characteristics of cells and tissues. Additionally, eosin's ability to stain the extracellular matrix can provide valuable information about the organization and composition of the surrounding tissue environment. The versatility and widespread use of eosin in various fields of biology and medicine make it a crucial topic of study and research.
3. Giemsa staining

Giemsa staining is a widely used technique in hematology and parasitology for the identification and differentiation of various cell types, particularly blood cells and parasites. The Giemsa stain is a complex mixture of methylene blue, eosin, and azure dyes, which collectively stain different cellular components with varying intensities, creating a distinctive color pattern that allows for the identification of specific cell types and their characteristics. This staining method is particularly valuable in the diagnosis and study of blood-borne diseases, such as malaria, where the identification of parasites within red blood cells is crucial for accurate diagnosis and treatment. Additionally, Giemsa staining is commonly used in cytogenetic analysis, where it can help visualize chromosomal structures and patterns, aiding in the detection of chromosomal abnormalities. The versatility and specificity of Giemsa staining make it an indispensable tool in various fields of biology and medicine, and its continued study and refinement are essential for advancing our understanding of cellular and molecular processes.
4. Romanowsky staining

Romanowsky staining is a widely used technique in hematology and cytology for the differential staining of various cell types, particularly blood cells. This staining method is named after the Russian physician Dmitri Leonidovich Romanowsky, who developed the original formulation in the late 19th century. The Romanowsky stain is a complex mixture of methylene blue, eosin, and other dyes that selectively bind to different cellular components, resulting in a distinctive color pattern that allows for the identification and differentiation of various cell types. This staining technique is particularly valuable in the analysis of blood smears, where it enables the visualization and classification of different types of blood cells, such as red blood cells, white blood cells, and platelets. Additionally, Romanowsky staining is used in the diagnosis and study of various hematological disorders, as it can provide valuable information about the morphological characteristics of blood cells, which can be indicative of underlying pathological conditions. The continued study and refinement of Romanowsky staining techniques are crucial for advancing our understanding of hematological processes and improving clinical diagnostic capabilities.
5. Giemsa staining of Malaria Blood films

Giemsa staining of malaria blood films is a crucial diagnostic technique in the identification and differentiation of malaria parasites, which are the causative agents of the infectious disease malaria. This staining method, which utilizes the Giemsa stain, a complex mixture of methylene blue, eosin, and azure dyes, allows for the visualization and characterization of the different stages of the malaria parasite's life cycle within the red blood cells of infected individuals. The distinctive color patterns created by the Giemsa stain enable the identification of specific malaria parasite species, as well as the differentiation of various developmental stages, such as trophozoites, schizonts, and gametocytes. This information is essential for accurate diagnosis, appropriate treatment, and epidemiological surveillance of malaria, a disease that remains a significant global health concern, particularly in tropical and subtropical regions. The continued study and refinement of Giemsa staining techniques for malaria diagnosis, along with the development of complementary diagnostic tools, are crucial for improving the detection and management of this infectious disease, ultimately contributing to global efforts to control and eliminate malaria.
6. G-banding

G-banding is a cytogenetic technique used for the visualization and analysis of chromosomal structures and patterns. This method involves the treatment of chromosomes with a combination of trypsin and Giemsa stain, which results in the formation of distinctive banding patterns along the length of the chromosomes. These banding patterns, known as G-bands, are unique to each chromosome and can be used to identify specific chromosomes, detect chromosomal abnormalities, and study the organization and structure of the genome. G-banding is widely used in various fields, including clinical genetics, cancer research, and evolutionary biology, as it provides a powerful tool for the identification and characterization of chromosomal aberrations, such as deletions, duplications, and translocations, which can be associated with genetic disorders, cancer, and other pathological conditions. The continued study and refinement of G-banding techniques, as well as the integration of this method with other cytogenetic and molecular techniques, are essential for advancing our understanding of the genetic basis of human health and disease, and for developing more effective diagnostic and therapeutic approaches.
7. Fixatives

Fixatives are chemical agents used in various fields of biology and medicine to preserve the structural and chemical integrity of biological samples, such as cells, tissues, and organisms, for subsequent analysis and observation. The primary function of fixatives is to prevent the degradation and alteration of the sample's morphology, composition, and molecular properties, ensuring that the sample accurately represents the in vivo state. There are a wide variety of fixatives, each with its own unique properties and applications, ranging from simple aldehydes like formaldehyde to more complex mixtures like Bouin's solution and Carnoy's fluid. The choice of fixative depends on the specific requirements of the study, the type of sample, and the desired analytical techniques. The proper selection and application of fixatives are crucial in fields such as histology, cytology, electron microscopy, and molecular biology, where the preservation of the sample's structural and chemical integrity is essential for accurate analysis and interpretation. Ongoing research into the development and optimization of fixatives, as well as the understanding of their mechanisms of action and their effects on different types of samples, is essential for advancing various areas of biological and medical research.