![]() ![]() ![]() Transgenic mice were injected with LiCl (0.15 M, 40 ml/kg, i.p.). In order to confirm mRNA expression and preservation in β-Gal-positive cells, we performed RT-PCR after LCM of single cells. ![]() Reference Kwon and Houpt 2010 PMID 19925827 RT-PCR was performed by using the OneStep RT-PCR Kit (Qiagen, Valencia, CA). Total RNA was extracted from individual LCM samples with the Total RNA Microprep Kit (Stratagene, La Jolla, CA) according to the manufacturer’s protocols. ![]() Following dissection, tissue sections were stained with methyl green (Vector Laboratories, Burlingame, CA) and coverslipped to verify the specificity of the dissection. Multiple laser shots (150-200) were required to transfer each amygdala region. LCM parameters were 15 or 30 µm diameter laser spot, a 10 ms laser pulse duration, and a 65 mW laser power. The frozen brain sections were defrosted at room temperature for 30 sec, then stained and dehydrated by the following procedure: 75% ethanol for 30 sec, DEPC-treated deionized H2O (DEPC-dH2O) for 30 sec, Histogene Staining solution (Arcturus, Mountain View, CA) for 3 min, DEPC-dH2O for 30 sec, 75% ethanol for 30 sec, 95% ethanol for 30 sec, 100% ethanol for 30 sec, xylene for 5 min, air dried in a fume hood for 15 min, and then desiccated with Drierite (W.A. The frozen brains were sectioned between -2.30 and -3.14 mm from bregma (Paxinos and Watson, 1986) at 5 µm thickness and mounted on slides in a –20 ✬ cryostat using the CryoJane system (Instrumedics, Hackensack, NJ). See also Electron Microscopy Sciences page LCM0213 caps and alignment tray and incubation block (dont know if we need all that)Īdhesive-Coated Slides CFSA 1X Slides White (125/box) item # 39475208 $375Īdhesive Tape Windows 1x3 Slides (RL200per) item #39475214 $114.2.2 Cryo-Jane System for Adhesive SlidesĮxample Powerpoint with videos and calibration images. ![]()
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