Sample Cages

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. We have found it best to put the samples into the cages after the SHIELD step or after delipidation. Please note that if you intend to actively delipidate your samples, you must first use the Batch Cage since that is the only cage that fits into the Clearing Cup. 

To insert the samples, first make a note of the letter on the segment of the 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 they cannot float out between the cage bars it will be alright to use. 

Once all samples have been loaded, cover up the cage with the appropriate lid and proceed to incubate. The lid prevents the samples from floating out of the top of the cages.

To label a single mouse brain or sample, remove the lid and attach the Single Cage Lock Ring to the top before attaching to the sample cup. If your sample floats, you can leave the lid on the cage. 

To label 2 samples, put one into the bottom "A" cage and cover it with the "B" cage. After stacking the cage, put the second sample into the "B" cage and drop into the cup. Then attach the Double Cage Lock Ring and insert in the sample cup. You may need to add more buffer to the sample cup to ensure the top sample 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. Then push the Medium Lock Ring onto the pegs of the top cage with the clips on the end facing down.  Insert the cages into the sample cup with the prongs on the lock ring above the cutouts on the sample cup surface. Once inserted, push the cage down as far as it will go.  You will likely 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 Batch Cup. (Or if you are delipidating, you will need to use the Clearing Cup independent of the number of samples). First insert some brains into the bottom "A-F" cages, then stack the second cage on top. Then slide the Batch Cage Lock Ring onto the top cage with the prongs facing down. Insert the cages into the sample cup with the prongs on the lock ring above the cutouts on the sample cup surface. Once inserted, push the cage down as far as it will go.  You may need to add more buffer to the sample cup to ensure the top level samples are fully submerged. 

To remove the samples from the cages, first remove the lid. For Single Cages, simply invert the cage over a petri dish or conical tube to remove the sample. To remove samples from the Medium or Batch Cages, first attach the Sample Removal Lid to the top side of the cage. This lid has an opening above a single slot. Make note of which slot is open and invert the cage to remove the sample.