presents Protocols

Category: BACs Bacterial Artificial Chromosome Protocols

Protocol presents the amplification of insert end sequences from bacterial artificial chromosome clones using TAIL-PCR. The amplified products are suitable as probes for chromosome walking and genome mapping and as templates for direct sequencing. The protocol has been used in rice genome studies. - [Read Amplification of Insert End Sequences from BACs Clones by Thermal Asymmetric Interlaced PCR]

Category: Microscopy Protocols: Fluorescence Microscopy Protocols

Article presents an introduction to fluorescence microscopy. Includes: Fundamentals of Excitation and Emission; Stokes' Shift; Fading, Quenching, and Photobleaching; Fluorescence Light Sources; Filter Terminology; The Fluorescence Light Budget; Detecting Single Molecules. - [Read Introduction to Fluorescence Microscopy]

Category: Immunology: Mononuclear Cell Isolation Protocols

Presents two methods for preparing dendritic cells (DCs), a highly specialized type of antigen-presenting cell (APC). The first method involves the isolation of DCs from mouse spleen, resulting in a cell population that is highly enriched in accessory cell and APC function. A support protocol for collagenase digestion of splenocyte suspensions is described to increase the yield of dendritic cells. The second method involves generating large numbers of DCs from mouse bone marrow progenitor cells. - [Read Isolation of Dendritic Cells Protocol]

Category: Cell Biology and Cell Culture: Apoptosis Protocols: Apoptosis Analysis

Measurement of Apoptosis and Other Forms of Cell Death. Jagan Muppidi, Melissa Porter, and Richard M. Siegel. As programmed cell death (PCD) or apoptosis has emerged as an important regulator of development and homeostasis in multicellular organisms, methods to quantify apoptosis and to distinguish it from necrosis have been developed. This unit presents a set of assays for these purposes, many of which are technically very simple and ideally suited to the study of hematopoietic cells. - [Read Measurement of Apoptosis and Other Forms of Cell Death]

Category: Microscopy Protocols: In Vivo Imaging Protocols

Sophisticated fluorescence microscopy methods & equipment, now allow cellular events to be studied at high resolution in living material. The studying of living fly tissues presents unique difficulties in keeping the cells alive, introducing fluorescent probes, & imaging through thick hazy cytoplasm. This protocol outlines the preparation of major tissue types amenable to study by time-lapse cinematography and different methods for keeping them alive. - [Read Time-Lapse Cinematography in Living Drosophila Tissues: Preparation of Material]