Prototype Sample Cages

Materials

Single Mouse Brain Cages:

2x "A" Cages
2x "B" Cages
4x Lids

Medium Sample Cup Mouse Brain Cages:

"A-C" Bottom Cage
"D-F" Top Cage
Lid
Sample Removal Lid

Batch Sample Cup Mouse Brain Cages:

"A-F" Bottom Cage
"G-L" Top Cage
Lid
Sample Removal Lid

How To Use

These sample cages are intended to be used as a replacement for the nylon mesh bags to hold samples. We have found that the mesh bags may contribute to additional nonspecific signal on the surface of the samples during labeling, or can damage the surface of softer samples. As of now, the cages are still prototypes and the material is not fully compatible with the higher temperatures and pH of Delipidation Buffer. So, the samples must still be handled as normal through the delipidation step in the protocol.

Proceed through the Delipidation step of the protocol.
Before pre-incubating your samples in Radiant Buffer or Primary Sample Buffer for immunolabeling, they can be loaded into the cages.
Make a note of the letter on the segment cage before using a spoon to drop the sample into it. For mouse brains it is best to place them cerebellum down to avoid damage to the olfactory bulbs. Other organs have been tested, including mouse lungs and hearts. As long as the samples are small enough so that they cannot float out between the cage bars it will be alright to use.
After all samples have been loaded, cover up the cage with the appropriate lid and proceed to incubate.
Setup the device as normal for labeling, and prepare your sample cup with the appropriate buffer and antibodies. If you wish to label 2 mouse brains, you can use the smallest sample cup, and stack the cages as shown in the picture above. However, make sure to remove the lid from the bottom cage before stacking. Before adding the cages to the sample cups, it is best to remove the Lids from the top layer of the cage. It is also likely that you will need to add more solution to the sample cup to ensure that all of the samples are fully submerged. See the images below for some examples of how the cages should look when in the sample cups.
Run the labeling as normal, but add the lids back on for the wash steps outside the device to prevent the samples from floating out.
After labeling is complete, you can remove the samples by putting on the Sample Removal Lid and inverting the cage above a conical tube or petri dish. The sample in that cage will fall out while the others remain inside. To remove the others simply reposition the Removal Lid and repeat.

To label a single mouse brain, it is best to cover a single cage with the lid and drop it into the sample cup as is.

To label 2 brains, put one into the bottom "A" cage and cover it with the "B" cage. After stacking the cage, put the second brain into the "B" cage and drop into the cup. You may need to add more buffer to the sample cup to ensure the top brain is submerged. Because of this high liquid level, it is best to remove the lid from the "B" cage before labeling.

If you have 3-6 mouse brains, you will need to use the Medium Cup. First insert some brains into the bottom "A-C" cages, then stack the second cage on top. Add the samples to that cage and insert into the sample cup. It is best to remove the lid from the top cage, since the sample level is high. You may need to add more buffer to the sample cup to ensure the top level samples are fully submerged.

If you have 7-12 mouse brains, you will need to use the BatchCup. First insert some brains into the bottom "A-F" cages, then stack the second cage on top. Add the samples to that cage and insert into the sample cup. It is best to remove the lid from the top cage, since the sample level is high. You may need to add more buffer to the sample cup to ensure the top level samples are fully submerged.

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