Download Oogenesis: Methods and Protocols by Ioannis P. Nezis PDF

By Ioannis P. Nezis

This quantity offers tools and methods to review oogenesis in a wide diversity of organisms, from crops to mammals. Oogenesis: equipment and Protocols courses readers via protocols on versions of developmental biology, oogenesis in vegetation, worms, fruit flies, mosquitos, butterflies, starfish, zebrafish, frog, fowl and mouse. Written within the hugely winning Methods in Molecular Biology series structure, chapters contain introductions to their respective themes, lists of the mandatory fabrics and reagents, step by step, without difficulty reproducible laboratory protocols, and tips about troubleshooting and fending off recognized pitfalls.

Authoritative and state-of-the-art, Oogenesis: equipment and Protocols goals to make sure winning leads to the additional research of this important field.

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The super-resolution microscopy core facility at Oslo University Hospital is acknowledged for access to a DeltaVision OMX microscope. References 1. Fuller MT, Spradling AC (2007) Male and female Drosophila germline stem cells: two versions of immortality. Science 316(5823): 402–404. 1140861 2. Spradling A, Fuller MT, Braun RE, Yoshida S (2011) Germline stem cells. Cold Spring Harb Perspect Biol 3(11):a002642. a002642 3. Eliazer S, Buszczak M (2011) Finding a niche: studies from the Drosophila ovary.

Other fixatives such as methanol can also be used, although FA is recommended for most stainings. Methanol is a precipitating fixative and causes denaturation of proteins, which might help when staining with antibodies directed toward denatured epitopes (see Note 14, Table 1 and Fig. 2). 6. Feeding the females with yeast paste and adding males promotes optimal oogenesis and ensures continued egg production. 7. The vials from which the female flies are collected should not be overcrowded during larval development to make sure that the quality of the ovaries is good.

Peters and Celeste A. 0 [39] 40 min Oil on coverslip Halocarbon oil (Voltalef 10S) [25] Culturing and Live Imaging of Drosophila Egg Chambers 43 Fluorescent marker(s) used NA NA NA (brightfield only) NA (brightfield only) NA GFP::Exu Rhodamine-labeled tubulin GFP-labeled nanos mRNA mito-GFP, mitoTracker GreenFM CY2-Gal4 > UAS-GFP::Moesin rhodamine-labeled tubulin, Oli-Green DNA dye; Polo::GFP slbo-, Actin < flip out > -, or upd-Gal4 > UAS-GFP::Moesin, UASactin::GFP, UAS-mCD8::GFP, UAS-dsRed::N; slbo-actin::GFP slbo-Gal4 > UAS-mCD8::GFP; Ubi-NLS::GFP Ubi-NLS::GFP, Ubi-EB1::GFP, α4-tub-Tau::GFP NA (brightfield only) tubP-Gal4 > UAS-Wcd::GFP or UAS-RFP::Wcd; H2B::RFP, Armadillo::GFP References [6] [20] [7] [8] [9] [23] [10] [30] [27] [21] [11] [35] [37] [13] [28] [25] Leica DM IRBE with Ultraview spinning disk (CSU10) Zeiss Stereolumar (live) or Zeiss Axiophot with QImaging RETIGA 1300 camera (DIC) Deltavision deconvolution microscope (Olympus IX70 with Coolsnap HQ CCD camera) Zeiss 510 Meta LSM confocal Zeiss Axioplan2 with Axiocam MRm camera Deltavision deconvolution microscope BioRad MRC600 or Zeiss 510 Meta LSM confocals Leica DM IRE with Ultraview spinning disk Zeiss 510 Meta LSM confocal BioRad MRC600 confocal on Nikon DiaPhot BioRad MRC600 laser confocal Laser ablations on Olympus BH2 Zeiss 4FL fluorescence microscope Leitz microscope and Bolex H16 Reflex Camera Undescribed light microscope (no live imaging) No imaging attempted Live imaging system(s) 5s 10 min 20 min 5 min 2 min 54 s 3–20 min 2–3 s 1 min 10 s Unspecified NA NA 100 s NA NA Interval Table 3 A chronological summary of 40 years of live-imaging efforts using the D.

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