Kubke Lab:Research/CND/Records/RC004

=Embryo details=

Species: Gallus gallus domesticus Embryo Name: RC004 Embryo stage: ST25 Staging description: Approximation, unconfirmed, Fixation: PFA Cryoprotection: None. Material label and storage: Slides in slide folder labeled RC004.

=Experiment details=

Objective: To investigate the best cryostat sectioning protocol. Use the tissue to establish Cresyl Violet staining protocol. Procedure: Comments: Sections from slide 5 were deemed by Fabiana to be sufficient for study. The settings used to cut slide 5's sections were:
 * Embryo staged under a dissection microscope in .9% saline solution. See Notebook entry.
 * Embryo loaded into a plastic mould filled with OCT and incubated in the cryostat chamber at -17°C.
 * Embryo loaded onto chuck, embedded deeper in OCT and incubated in the metallic chuck holder at -17°C for 30minutes.
 * Sectioned the embryo whilst modifying the settings of the cryostat to obtain the best sections.
 * The sectioned were mounted onto polylysine slides and dried overnight.

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 * Cryostat = Leica – CM3050S
 * Knife = MX35 Premier +, 34 degrees, 80mm Thermo Scientific
 * Day Cut = 17/12/2010, 12:30pm (see Notebook entry)
 * Knife Angle = 1.5°
 * Chamber Temp = -19°C
 * Object Temp = -19°C
 * Glass Slides = Polylysine subbed slides.
 * Plane of section = Coronal
 * Number of slides = 1
 * Observations = Was cutting the sections very quickly. Histology looked good under a microscope.

Cresyl Violet staining For more informattion see Kubke_Lab:Nissl_Stain_Protocol. Slides 1-5 were all stained together in one staining rack.

=Results=

Sections were stained too intensely. They require less heavy staining. The results of this study were used to perform cryostat sections on following embryos.

=Images= =Summary=