Sample Mounting

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(Place sample onto glass dish)
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Using a spatula, carefully place the tissue sample on the center of the dish surrounded by, but not touching, the putty. (A mouse brain before clearing is shown for the purpose of demonstration.)
 
Using a spatula, carefully place the tissue sample on the center of the dish surrounded by, but not touching, the putty. (A mouse brain before clearing is shown for the purpose of demonstration.)
 
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File: Placing_sample_on_dish.jpg|Placing tissue sample on the glass dish
 
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Revision as of 06:39, 16 April 2014

Before mounting the sample for imaging, incubate the sample in the mounting solution for several hours or overnight (until the sample is visually transparent. Suggested methods for mounting large tissue samples (whole brain) and smaller sections (<1 mm thick brain slice) using the imaging chamber supplies are detailed below.

Contents

Whole brain mounting for confocal imaging

Form a putty cylinder

Roll a piece of BluTack putty into a cylinder with uniform thickness. The cylinder should be several centimeters in length with a thickness greater than that of the brain sample.


Place putty on a glass dish

Shape the putty cylinder into a horseshoe U shape. Place the putty onto the flat side (outside) of a Willco glass dish. Using a pipette tip, press the outside edge of the putty U down onto the glass dish to help seal it.


Place sample onto glass dish

Using a spatula, carefully place the tissue sample on the center of the dish surrounded by, but not touching, the putty. (A mouse brain before clearing is shown for the purpose of demonstration.)


Sandwich sample with second glass dish

Add FocusClear to inside of chamber

Apply silicone elastomer to seal chamber

Thin slice mounting for confocal imaging

Incubate sample in FocusClear

Form a thin putty cylinder

Place putty on glass slide

Place sample on glass slide between putty

Sandwich sample with glass dish/cover slip

Add FocusClear to inside of chamber

apply silicone elastomer to seal chamber