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This document discusses flow cytometry and its application in measuring CD4+ T lymphocytes. It describes how flow cytometry works using a laser to detect cell properties like size, granularity and fluorescent markers. The BD FACSCalibur flow cytometer is highlighted as an example system used to quantify CD4+ T cells. The procedure involves staining blood samples with fluorescent antibodies and analyzing the results to determine CD4+ T cell counts and percentages, which are important markers for HIV/AIDS monitoring and treatment. Normal reference ranges are provided.
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The document discusses flow cytometry and its clinical application in monitoring CD4 T lymphocyte counts. Flow cytometry works by passing fluorescent-labeled cells in a fluid stream through a laser which causes fluorescence. Detectors then measure the cells' light scattering and fluorescence properties to characterize the cells and identify subsets. The document provides details on using the BD FACSCalibur flow cytometer to measure CD4 counts via two-color staining and gating on T lymphocyte populations. Normal CD4 values in adults and children are listed.
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The document is a 17 page paper on flow cytometry. It begins with an introduction and overview of flow cytometry, including its basic concepts, parameters measured, and how fluorescence is involved. It then reviews a research paper on microfluidic flow cytometry, summarizing the paper's introduction, theory, design of experiment, fabrication, results, and conclusions. The paper achieved over 94% accuracy compared to flow cytometry in determining the percentages of CD4+ and CD8+ T cell subpopulations in mixed samples. It demonstrates the potential for a microchip-based system to perform label-free immune cell analysis in real-time. In the future, the measurement throughput could be improved by revising the fluidic design to lower hydraulic resistance
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This document provides an overview of flow cytometry, including its principles, instrumentation, applications in hematology and oncology, and techniques for analyzing results. Flow cytometry works by passing single cells in a fluid stream through a laser beam, which detects cellular properties like size, structure, and antigen expression. It allows for multiparametric analysis of up to 12 parameters simultaneously on thousands of cells. Key applications include immunophenotyping of leukemias and lymphomas, DNA analysis, and monitoring of minimal residual disease.
FLOWCYTOMETRY........................pptx
Flow cytometry allows for the quantitative and qualitative analysis of physical and chemical characteristics of cells as they flow in a fluid stream through a laser beam. It measures properties like cell size, granularity, and fluorescence. The main components are the fluidic system that orders cells into a single-file stream, the optical system that applies light and collects signals, and the electronic system that converts signals to digital data. As cells pass through the laser, they scatter and fluoresce light that is detected and analyzed to characterize cell populations and identify abnormal cells. It is used for applications like leukemia diagnosis and monitoring transplant engraftment.
cell-counters1575.ppt.pptx
Wallace Coulter developed the Coulter Principle for counting and sizing microscopic particles like blood cells in the late 1940s-early 1950s. Modern cell counters use various technologies including impedance, absorbance spectrophotometry, optics, fluorescence, conductivity, and monoclonal antibodies. Cell counters must run within specifications to provide accurate results.
Cell Counter CBC New.pdf
Wallace Coulter developed the Coulter Principle for counting and sizing blood cells using impedance technology in the late 1940s. Modern cell counters use various technologies including impedance, absorbance spectrophotometry, optical light scattering, fluorescence, radio frequency, and monoclonal antibodies to provide complete blood cell counts, differentials, reticulocyte counts, and other parameters from small blood samples. Cell counters must operate within quality control standards to provide accurate and precise results.
Detection of somatic mutations at 0.5% frequency from cfDNA and CTC DNA using...
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1) The document describes a multiplex next-generation sequencing assay that can detect somatic mutations at a frequency of 0.5% from circulating tumor cells (CTC) and cell-free tumor DNA (ctDNA) obtained from a single blood sample.
2) A research workflow is presented that isolates white blood cells, plasma DNA, and CTCs from blood samples, prepares barcoded libraries from each fraction, and sequences them on an Ion chip to enable detection of low frequency variants down to 0.5% using statistical modeling and filtering.
3) The assay requires only 10ng of input DNA, has a turnaround time of less than 24 hours, and demonstrates the ability to accurately detect somatic mutations atflow cytometry by aksharaditya shukla.
Flow cytometry measures properties of cells in a fluid stream by staining cells with fluorescent antibodies and analyzing them with a flow cytometer instrument. It allows for the characterization of cell populations based on cell surface and intracellular markers. Flow cytometry is useful for immunophenotyping in diseases like leukemia where it can identify the cell lineage involved (lymphoid vs myeloid) and detect minimal residual disease during treatment. In this case, the 5-year old boy presenting with fever, flow cytometry immunophenotyping should be performed to diagnose causes like acute lymphoblastic leukemia by examining markers like CD19, CD5, CD10 on lymphocytes.
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ROLE OF FLOW CYTOMETRY IN LEUKEMIAS
1) Flow cytometry is used to measure multiple physical and chemical properties of cells in a fluid stream at a rate of thousands of cells per second. It is used to diagnose and classify leukemias based on antigen expression.
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3) Flow cytometry utilizes light scattering and fluorescence to identify cell size, granularity, lineage, and maturation stage based on antigen expression. This immunophenotyping is essential for diagnosing and distinguishing between different types of leukemias.
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FlowBasics2[1]
Flow cytometry is a technique that allows for the measurement of physical and chemical characteristics of single cells flowing through a stream of fluid. It enables the simultaneous multiparametric analysis of thousands of cells per second. Cells are labeled with fluorescent markers and passed individually through a laser beam, where detectors measure the cells' light scattering and fluorescent properties. This information can be used to identify and sort cell subpopulations. Flow cytometry provides high-speed analysis of multiple parameters at a single-cell level and is widely used in research, clinical diagnosis, and other applications.
Flowcytometry
Flow cytometry plays an indispensable role in the diagnosis of hematological disorders by providing data on immunophenotype. It is a method that can measure multiple characteristics of single cells using fluorescent markers and lasers. This allows determination of cell size, granularity, protein expression and more. Two cases are described where flow cytometry was used. In case 1, a leukemia sample showed expression of markers consistent with acute promyelocytic leukemia. In case 2, a mediastinal mass sample expressed markers indicating acute T-cell leukemia. Flow cytometry provides valuable immunophenotyping data for diagnosis of hematological malignancies.
Basics of Flow cytometry by Prof. Mohamed Labib Salem
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みなさんこんにちはこれ何文字まで入るの？40文字以下不可とか本当に意味わからないけどこれ限界文字数書いてないからマジでやばい文字数いけるんじゃないの？えこ...
みなさんこんにちはこれ何文字まで入るの？40文字以下不可とか本当に意味わからないけどこれ限界文字数書いてないからマジでやばい文字数いけるんじゃないの？えこ...
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- 7. Flow CytometerBD FACSCalibur™ System 7
- 8. 8
- 9. Sistem Fluida Terdiri dari sheath fluid dan central channel Tenaga hidrodinamik sel satu persatu melewati central channel Fluidics–Hydrodynamic Focusing Excitation Optics (laser) 9
- 11. Mendeteksi sinar pada sudut 900
- 14. BD FACSCalibur Optics 11
- 15. 12
- 16. Sistem komputer Pengumpulan dan analisis data : mengubah sinyal analog sinyal digital analisa tegangan pulsa : tinggi, luas, lebar 13
- 17. Prosedur Pemeriksaan Limfosit CD4+ Dengan Menggunakan Flow Sitometer BD FACSCalibur™ System Prinsip : Tabung reagen mengandung antibodi yang ditandai fluorochrome dalam jumlah tertentu. Jika ditambahkan sampel, antibodi tsb akan terikat dengan antigen pd permukaan limfosit dan akan berfluoresensi bila terkena sinar laser. Fluoresensi yang terjadi sebanding dgn jumlah sel yang ada 14
- 18. 15
- 20. Cara kerja 17
- 22. Limfosit T CD4 % : ≥ 25%
- 23. Bayi ≥ 12 bulan :
- 24. Limfosit T CD4 absolut : ≥1500/cmm3
- 26. 1 – 5 tahun :
- 27. Limfosit T CD4 absolut : ≥1000/cmm3
- 28. Limfosit T CD4 % :≥25%18
- 29. Contoh printout pemeriksaan limfosit T CD4+ menggunakan Flow Cytometer BD FACSCalibur Nilai absolut dan prosentase limfosit T CD4+ yang sangat menurun 19
- 32. Terima Kasih 22
- 33. Current CD4 monitoring tests and technologies include Flow based assays Becton Dickinson FACSCount Guava EasyCD4 PartecCyFlow Beckman Coulter PointCARE Manual immune bead-based assays DynalDynabeads Coulter Cytosphere In the pipeline LabNow microchip technology SemiBio slide test (microscopy) 23
- 34. T – cell subsets CD3 T CD3 CD3 T T C H CD4 CD8 24
- 35. Specific Antibody Antigen Foreign Invader 25
- 36. Incubate Two-Color Direct Staining Analyze 26
- 37. 27
- 40. Detected at 90° to the laser beam28
- 41. largest and most complex population 1000 Neutrophils 800 600 400 Monocytes 200 smallest and least complex population Lymphocytes 0 0 200 400 600 800 1000 Lysed Whole Blood Side Scatter Forward Light Scatter 29
- 43. Menghitung Σ absolut CD4 Dengan membandingkan “cellular even” terhadap “beads even” Pada BD Multiset : otomatis Cara manual : Absolut count of cell = Σ of events in regio Σ of beads per test containing cell X Σ of events in absolute test volume count bead region 31
- 45. 33
- 46. 34
- 47. 35