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FocusClear™ Product Demonstration

(A) Materials and devices for sample clearing and embedding: 5 ml each of FocusClear™, MountClear™ and Immersion Solution-M, a 24-well cleaning plate, cleaning paper, a cover slip, reinforcing rings, a loop and a clamp.

(B) Before to begin sample clearing, put a few drops of FocusClear™ (about 100 μl) into as many wells of a 24-well plate as needed. If the thickness of specimen is over 400 microns, using an epen droff as the incubation chamber is recommended.
(C) Next, transfer the samples into the wells using a micro-loop.
(D) Then, cover the wells tightly with a tape to avoid dehydration.
(E) Cover the whole plate with a tinfoil to avoid photo bleaching in the samples. Keep the plate at room temperature for the desired incubation period.
(F) Before to begin samples embedding, prepare a slide and clear it with a piece of tissue.
(G) Then, build a chamber on the slide by putting pieces of reinforcing rings together. Approximately 2 pieces of reinforcing rings will be needed to build a chamber for a specimen about 200 microns thick.
(H) Put a few drops of MountClear™ (10-20 μl) into the chambers and then transfer the FocusClear™ treated samples from the well into the chambers.
(I) Cover the chamber with a cleaning cover slip, and slightly press the edge of the cover slip to squeeze out any redundant MountClear™ solution.
(J) Wipe away the unwanted MountClear™ solution, and then seal the chambers by putting on nail polisher around the edge. Wait for the nail polisher to dry.
(K) As the nail polisher is dried and the edge of the chamber is sealed, your samples are now ready to be viewed under a confocal microscope. When using an oil/water immersion lens to observe the sample, Immersion Solution-M matching the reflective index of the mounting solution should be used for better resolution.
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