Temperature. K Heads K in 0.1 M cacodylate buffer dehydrated in a series of ethanol inwater w Deleted with propylene oxide of 100%, and Equilibrated in Spurr resin quilibrierungsschritt s propylene oxide in Spurr resin Each view is rotated is set to 8 carried out at Amonafide AS1413 room temperature. KK Heads were placed in molds and cured overnight at 70. The eyes were cut with a Reichert Jung microtome ULTRACUT E. Ultrathin sections were compared to grid and uranyl acetate and lead citrate PIOLOFORM onehole Reynolds 8% in ethanol 50%. The products were examined Philips CM120 electron microscope. Quantification of the genotype in neurodegenerative Ph show the number of morphologically normal photoreceptor neurons Thurs field size S e occurs in the middle of the eye. At least four individual eyes and 260 ommatidia were scored for each genotype.
Ultra Thin regions were at a constant depth of 25 m from the surface Surface of the eye made. Analysis software was used to create multiple overlapping images of the eye area BMS-387032 only ? BEP mag BEP were to make of 2050, compiled with Adobe Photoshop. Rhabdomeres were considered normal if it contains Lt no books and L are not condensed, fragmented or divided. P values were calculated with a fa Here or two-way ANOVA test and Dunnett’s multiple comparison or Bonferroni post-test calculated with Prism 4.0c software. For immunofluorescence Immunofluorescence antennal imaginal eyes from wandering third instar larvae were dissected in PBS and 1 ? PH3, BrdU, TUNEL and acridine orange as described. The major organs were old Ren PH3, Elav, BrdU and DIG conjugated rhodamine.
Fluorescent conjugates of the old corpses were secondary Ren Ren mouse FITC rabbit FITC, rhodamine rats and rats TUNEL Alexa Fluor 488 tests were used with the kit Red Apoptag apoptosis in situ detection. Following changes to the log F Wolff TUNEL-F was carried out coloring: Was performed 1 ? PBS was replaced by 0.1 M sodium phosphate buffer replaced adjusting ngliche anf 5% formalin / PBS for 1 ? eye were discs were introduced into the reaction mixture for 1-37 h incubated to the block has been stopped at room temperature for 1 l L solution in an L solution of 12% block length length / TX BSS/0.3 done conjugated rhodamine 100% and stirred DIG old K body were ELAV BS / BSS / TX diluted and eye discs were incubated overnight at 4 and rat Alexa Fluor 488 was diluted in BS / BSS / TX and eye discs were incubated overnight at 4, and described by Wolff treated.
1 ? PBS was replaced by Ringer’s solution L Drosophila: The following amendments to the Protocol acridine orange staining F F Wolff achieved. Eye antennal imaginal discs were imaged on a Zeiss Axiovert 200M inverted microscope. FACS FACS analysis was performed as described by Neufeld et al. with the Ver changes specified directory. Ante Nalen imaginal m Moderately 3-40 Gl Migrate these third instar larvae were washed in PBS pr and immediately transferred to 500 TEHP buffer L. Eye antennal imaginal discs were withgentle agitation was incubated for 3 h. The use of differentiated cells were 10 times and the discs gently pipetted every 20 minutes. After dissociation, 100 L of 33 g / ml propidium iodide in PBS was added to each sample. A Becton Dickinson LSRII was used to analyze the cells. Five
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