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This document discusses the preparation and experimental protocol for studying isolated blood vessels, specifically the middle cerebral artery. Key points: - The system allows for the isolation of small blood vessels and study of their structure and response to various pharmacological and physiological stimuli. - Phosphate buffered saline (PSS) solutions are prepared to properly maintain and superfuse the isolated vessels. - Middle cerebral arteries are isolated from rats using anesthesia, decapitation, and incubation in oxygenated PSS solution. - An experimental protocol applies increasing pressure gradients and tests responses in the presence or absence of various inhibitors and vasoactive drugs. - Statistical analysis of data uses repeated measures two-way ANOVA.
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The document summarizes the process of preparing tissue samples for histological analysis, including fixation, dehydration, infiltration/embedding, sectioning, staining, and examination. Key steps involve fixing tissues to prevent degradation, dehydrating using increasing alcohol concentrations, infiltrating with paraffin wax or resin for structural support during sectioning, precisely cutting thin sections, mounting them to glass slides, staining, and examining under a microscope. The quality of prepared samples depends on carefully following each step of the preparation process.
Notes for The principle and performance of capillary electrophoresis
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Notes for The principle and performance of liquid chromatography–mass spectro...
This document provides an overview of liquid chromatography-mass spectrometry (LC-MS). It describes the basic components and functioning of an LC-MS system, including the liquid chromatograph and mass spectrometer connected by an interface. The document discusses various ionization sources like electrospray ionization and atmospheric pressure chemical ionization, as well as mass analyzers like quadrupoles and time-of-flight analyzers. It also covers detectors used in LC-MS like electron multipliers and photomultipliers. Overall, the document serves as a technical introduction to the principles and components of LC-MS.
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This document provides an overview of basic cell culture techniques. It discusses the history of cell culture, defining primary and secondary cell cultures. It describes different types of cell lines and how cells grow as monolayers or in suspension. The document outlines the key equipment needed for a cell culture laboratory, including biosafety cabinets, CO2 incubators, centrifuges, microscopes, and supplies. It emphasizes the importance of aseptic technique to prevent microbial contamination when working with cell cultures.
Big datasets
This document discusses systems biology and its goals of understanding how biological molecules interact and systems function as a whole. It covers:
1) Systems biology uses large datasets from "omics" experiments and computational models to understand complex biological interactions beyond individual molecules.
2) Pioneering work used microarrays to measure thousands of genes in serum-stimulated cells, finding over 500 changed in proliferation.
3) The field aims to discover emergent system properties and functions not evident from separate parts, like switches that change cell behavior.
Systems biology for Medicine' is 'Experimental methods and the big datasets
This document discusses experimental methods used in systems biology to generate large datasets, including microarrays, sequencing-based methods, mass spectrometry, and liquid chromatography. It explains that systems biology studies must be quantitative and enable computational modeling. Key methods covered are microarrays, RNA-seq, ChIP-seq, whole-genome sequencing, whole-exome sequencing, proteomics using mass spectrometry, and combining liquid chromatography with mass spectrometry for lipidomics, metabolomics and glycomics. Sources of variation are also discussed for genomic and proteomic studies.
Systems biology for medical students/Systems medicine
Systems biology takes a holistic approach to studying biological systems by considering all the interactions within a system and how they generate complex behaviors. Lecture 1 introduces key concepts in systems biology like how increasing levels of biological organization give rise to new system properties like robustness. Lecture 2 discusses experimental methods like genomics, proteomics, and metabolomics that generate large data sets for systems analysis. Lecture 3 covers mathematical and statistical tools for analyzing these data sets, such as using differential equations to model signaling networks. Lecture 4 provides examples of medical applications of systems biology in finding diagnostic markers, personalizing therapy, and predicting disease interactions from human disease networks, with the future of medicine taking a more predictive, preventive, and personalized approach
Use cases
The document discusses several use cases for applying data mining and machine learning techniques in healthcare and biomedical research. Three examples are:
1) Early diagnosis of cancers like lung cancer and breast cancer through predictive modeling of patient data to detect cancers at earlier stages when survival rates are higher.
2) Predicting patient responses to drug therapies for cancers like breast cancer by combining different types of molecular profiling data using techniques like support vector machines and random forests.
3) Using imaging data and temporal analysis of metrics like medication purchases to better understand and predict chronic diseases like diabetes and associated health complications.
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- 1. Isolated blood vessels Ivana Jukic
- 2. PARTS OF THE SYSTEM AND FUNCTION • proučavanje structure and isolated features blood small vessels diameter (> 60 microns) - rat middle cerebral artery • myogenic answer / Flow induced dilatation • različiti pharmacological / physiological stimuli • inconstructed the system heating • opens for superfusion. quickly discharge and filling (addition of drugs / oxidation) • cleaning and proper maintenance • danish Myo Technology (DMT) (Denmark) / MyoVIEW • flowmeter - 161FM • flow between 15-1500 uL/ min
- 3. Picture First Components of isolated blood vessels (pressure myograph System DMT Model 111P)
- 4. PREPARATION PSS SOLUTIONS PSS SOLUTION • For prepare 0.5 L • Distilled water: 450 mL • Salt Stock solution: 25 mL • buffer Stock solution: 25 mL • Glucose: 0.5 g • NaH2PER4: 0.07 Mr Ca-free PSS SOLUTION • To prepare 0.5 L • Distilled water: 450 mL • Salt Stock Ca-free solution: 25 mL • buffer Stock Ca-free solution: 25 mL • Glucose: 0.5 g • NaH2PER4: 0.07 g pH 7:35 to 7:45
- 5. INSULATION middle cerebral artery • Andnestezija • ketamine 75 mg / kg and midazolam 0.5 mg / kg • decapitation • isolation artery • PSS (37C) • toontinuirana oxygenation system • 21% O2, 5% CO2 and N balance2 • andnkubacija blood vessels • 60 min / Δ80 mmHg
- 6. EXPERIMENTAL PROTOCOL • basic values • oppressive gradients Δ10, Δ20, Δ40, Δ60 and 100 mmHg • prisutnost / absence inhibitor • L-NAME • indomethacin • MESP • TEMPOL • Ach / SNP • Ca2+- free PSS solution
- 7. STATISTICAL DATA AND PRESENTATION • data processing must use two-way ANOVA repeated measures Picture 2. The basic answer resistive arterioles subcutaneous (SAT) and visceral (VAT) adipose tissue to flow induced dilation (FID), comparison between the two groups