Suggested procedures for processing fixed tissue
Here are a few guidelines which you can use to process fixed tissue using Histo-Clear and Histo-Clear II:
Histo-Clear Automated Tissue Processing Schedule
|Process Bath||18 Hour Cycle (Time in Hours)||24 Hour Cycle (Time in Hours)|
Histo-Clear II Tissue Processing Protocol
|Process Bath||Time (Hours)|
|Anhydrous isopropanol (optional)||1.5 (may help to remove last trace of water)|
|10% ethanol, 90% Histo-Clear II||4|
Histo-Clear II Dewaxing and Staining Protocol
|Process Bath||Time (minutes)|
|10% ethanol, 90% Histo-Clear II||5|
For best results with Histo-Clear II:
- Rotate Histo-Clear II after every 3-5 runs for heavy use, or every 5-7 runs for light use.
- Replace the final absolute ethanol bath frequently, or at the first sign of water contamination. Histo-Clear II is very water intolerant. Including an isopropanol step before the first Histo-Clear II station and/or adding 10% ethanol to the first Histo-Clear II station will minimize the effects of water carryover (see protocols above).
- Mounting medium: Use a compatible medium, such as Omnimount (HS-110). Histo-Clear II is not compatible with DPX or Histomount.
Note: These protocols are recommended starting points for developing an optimal process for a given lab's equipment, tissue types, section thickness, etc.
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