Frozen block embedding |Frozen block embedding II |Frozen block embedding III
Frozen block cryoembedding
Small Skin Ellipse Click on thumbnails for larger pictures
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A) Place tissue on a thin film of
embedding medium face down on the dispensing slide and place the tissue on
the freezing griddle.
B) Cover the tissue with an ample quantity of embedding medium. C) Place the elevated freezing block over the tissue. Freezing time is about one minute. |
D) Remove elevated freezing block
with a twist. The lines show the cuts to be made in numerical order. E) Remove the cutting board to the workbench. Test the frozen block for hardness while trimming the edges of the frozen medium. The block should cut like hard ice cream. F) If there is any tendency for the cut piece to fly apart it is too hard. Warm the frozen block for a few seconds between the gloved hands and retest. |
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G) Cut the
frozen block into the appropriate sections to achieve the desired
orientation.
H) When cutting these longitudinal margins push the blade toward the medium and make sure the tissue surface is well frozen to the medium to prevent dislodging. I) Cut pieces shown face up. |
J) Place the frozen slices face down in the
embedding well using the cold epoxy coated forceps to prevent sticking.
Leave about 2mm. space between pieces for medium to penetrate.
K) Fill well place chuck and allow freezing then remove block. M) Block after removal from well. |
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N -0) Plastering
technique fills any imperfections on the surface.
Apply a drop of medium; press on any cold surface and remove with a tap. |
P) Trimmed block face. Q) Photograph of frozen section slide. R) Photomicrograph composite.(50 X magnification) |
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Very large skin ellipse Flounder on the griddle
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1) Large (6.5cm) ellipse of skin. Very thin measuring from 1 -2 mm thick. 2) Freezing on the griddle inked surface down. 3) Cut tissue into four portions. Take each quadrant one at a time to the cutting board outside of the cryostat. Each portion is cut into about 7-8 slices. Four slices from each quarter are embedded on edge in a large well. |
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4) Four blocks correspond to four quadrants. Each
is trimmed. All show full sections on edge with inked margins clearly
visible. 5) Frozen section slides in composite photo corresponding to four blocks and quadrants. Result is that over 1/2 of this very large specimen is examined in beautiful on edge sections including all deep and lateral margins by preparing only four blocks. |
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