Search results for "Cell analysis"
showing 10 items of 41 documents
Cell population analysis of the adult murine subependymal neurogenic lineage by flow cytometry
2021
Summary This protocol provides a flow-cytometry-based procedure to classify and isolate all cells of the adult rodent subependymal zone (SEZ) neurogenic lineage, without the need for reporter mice, into different cell populations, including three neural stem cell (NSC) fractions with molecular signatures that are coherent with single-cell transcriptomics. Additionally, their cycling behavior can be assessed by means of 5-ethynyl-2′-deoxyuridine (EdU) incorporation. Our method allows the isolation of different NSC fractions and the functional assay of their cycling heterogeneity and quiescence-activation transitions. For complete details on the use, execution, and outcomes of this protocol, …
Toward fast malaria detection by secondary speckle sensing microscopy
2012
Diagnosis of malaria must be rapid, accurate, simple to use, portable and low cost, as suggested by the World Health Organization (WHO). Despite recent efforts, the gold standard remains the light microscopy of a stained blood film. This method can detect low parasitemia and identify different species of Plasmodium. However, it is time consuming, it requires well trained microscopist and good instrumentation to minimize misinterpretation, thus the costs are considerable. Moreover, the equipment cannot be easily transported and installed. In this paper we propose a new technique named "secondary speckle sensing microscopy" ((SM)-M-3) based upon extraction of correlation based statistics of s…
Laser Microdissection of the Alveolar Duct Enables Single-Cell Genomic Analysis
2014
Complex tissues such as the lung are composed of structural hierarchies such as alveoli, alveolar ducts, and lobules. Some structural units, such as the alveolar duct, appear to participate in tissue repair as well as the development of bronchioalveolar carcinoma. Here, we demonstrate an approach to conduct laser microdissection of the lung alveolar duct for single-cell PCR analysis. Our approach involved three steps. The initial preparation used mechanical sectioning of the lung tissue with sufficient thickness to encompass the structure of interest. In the case of the alveolar duct, the precision-cut lung slices were 200µm thick; the slices were processed using near-physiologic conditions…
Assessment of the quality of the air in the city of Palermo through chemical and cell analyses on Pinus Needles
2001
The influence of air pollution on the chemical composition of Pinus sp. needles was examined in polluted and control sites in and around the city of Palermo (Sicily). The chemical composition of needles indicated the extent of contamination of the trees, which were cytologically examined. Cell analysis was carried out on pine samples, including needles and pollens, from 15 different locations. Biostructural and spectrophotometric tests were performed. In particular, concentrations of toxic (Cd, Pb) and non-toxic metals (Fe,Cu, Zn) were determined, as well as injury caused by their accumulation in the needles. The more highly urbanised areas showed higher concentrations of metals (Pb, Cu. Zn…
Cryopreservation of MHC Multimers: Recommendations for Quality Assurance in Detection of Antigen Specific T Cells
2015
Fluorescence-labeled peptide-MHC class I multimers serve as ideal tools for the detection of antigen-specific T cells by flow cytometry, enabling functional and phenotypical characterization of specific T cells at the single cell level. While this technique offers a number of unique advantages, MHC multimer reagents can be difficult to handle in terms of stability and quality assurance. The stability of a given fluorescence-labeled MHC multimer complex depends on both the stability of the peptide-MHC complex itself and the stability of the fluorochrome. Consequently, stability is difficult to predict and long-term storage is generally not recommended. We investigated here the possibility of…
Speckle based sensing device for fast detection of malaria
2012
We propose a new technique for malaria detection. It is based upon extraction of correlation based statistics of speckle patterns generated while illuminating red blood cells with a laser and inspecting them under a microscope.
Messenger RNA Sequencing of Rare Cell Populations in the Lung and Lung-Draining Lymph Nodes
2017
Next-generation sequencing (NGS) techniques provide unique prospects for in-depth transcriptome analyses. Nevertheless, the emerging and still growing knowledge about the large diversity and heterogeneity of cells that participate in immunological responses in a tissue- and micromilieu-specific manner calls for advanced isolation and sequencing methods for the accurate quantification of gene expression in small cell populations and even individual cells from any organ or tissue. One of the major limitations in performing transcriptome analyses of rare cell populations was and still is quality and quantity of RNA that often limits analyses of complex mixtures of immune cell populations. Here…
Prediction model for aneuploidy in early human embryo development revealed by single-cell analysis.
2014
Aneuploidies are prevalent in the human embryo and impair proper development, leading to cell cycle arrest. Recent advances in imaging and molecular and genetic analyses are postulated as promising strategies to unveil the mechanisms involved in aneuploidy generation. Here we combine time-lapse, complete chromosomal assessment and single-cell RT–qPCR to simultaneously obtain information from all cells that compose a human embryo until the approximately eight-cell stage (n=85). Our data indicate that the chromosomal status of aneuploid embryos (n=26), including those that are mosaic (n=3), correlates with significant differences in the duration of the first mitotic phase when compared with e…
Labeling of Single Cells in the Central Nervous System of <em>Drosophila melanogaster</em>
2013
In this article we describe how to individually label neurons in the embryonic CNS of Drosophila melanogaster by juxtacellular injection of the lipophilic fluorescent membrane marker DiI. This method allows the visualization of neuronal cell morphology in great detail. It is possible to label any cell in the CNS: cell bodies of target neurons are visualized under DIC optics or by expression of a fluorescent genetic marker such as GFP. After labeling, the DiI can be transformed into a permanent brown stain by photoconversion to allow visualization of cell morphology with transmitted light and DIC optics. Alternatively, the DiI-labeled cells can be observed directly with confocal microscopy, …
Cell degradation detection based on an inter-cell approach
2017
Fault management is a crucial part of cellular network management systems. The status of the base stations is usually monitored by well-defined key performance indicators (KPIs). The approaches for cell degradation detection are based on either intra-cell or inter-cell analysis of the KPIs. In intra-cell analysis, KPI profiles are built based on their local history data whereas in inter-cell analysis, KPIs of one cell are compared with the corresponding KPIs of the other cells. In this work, we argue in favor of the inter-cell approach and apply a degradation detection method that is able to detect a sleeping cell that could be difficult to observe using traditional intra-cell methods. We d…