Plant Tissue Optical Clearing Reagent

Store at room temperature for up to one year.

During storage or transport at low temperatures, precipitation may occur in this product. In such a case, warm the product in a 37°C water bath or incubator until the precipitate dissolves completely. This does not affect its performance.
This product requires a relatively longer time for tissue clearing. For plant materials or dyes without specific requirements, it is recommended to perform clearing before staining. For faster clearing, use the BeyoFast™ Plant Tissue Optical Clearing Kit (C0468) that requires just 2-3 days for tissue clearing.
This product can also be used for clearing plant tissues after fluorescent staining, which does not interfere with fluorescence. However, the clearing process should be carried out in the dark to minimize fluorescence quenching.
Tender tissues may become fragile after clearing. Handle them gently with flat-tip tweezers to avoid damage.
The required amount of reagent should be enough to fully submerge the plant materials. Adjust the volume as needed based on sample size.
For your safety and health, please wear a lab coat and disposable gloves during the operation.

Tissue Clearing

1.Sample fixation
Collect plant samples, rinse 1-2 times with pure water, and fix at 4°C for 30-120 minutes (e.g., 30 minutes for seedlings, 120 minutes for leaves and roots, and 60 minutes for pistils).
Note: If necessary, apply moderate vacuum during fixation to enhance penetration of the Fix Solution.
2.Sample washing
Take the fixed samples and drain excess liquid. Wash twice with a Wash Buffer (e.g., C0221A) with gentle shaking on a shaker (e.g., BeyoShaker™ Digital See-Saw Rocking Shaker, E6673), at least 1 minute per wash.
3.Sample clearing
a.Submerge the samples in Plant Tissue Optical Clearing Reagent and clear at room temperature or 37°C for 4 days to 4 weeks (e.g., 4 days for leaves and roots, 7 days for seedlings, 2 weeks for pistils, and 4 weeks for mature tissues). Adjust clearing time as needed.
b.During clearing, if the clearing reagent changes from colorless to green, replace with fresh reagent until the plant sample becomes transparent. Cleared samples can be stored in the clearing reagent at room temperature for at least 3 months.
Note: If necessary, apply moderate vacuum during clearing to ensure thorough immersion and reduce clearing time. For even faster clearing, BeyoFast™ Plant Tissue Optical Clearing Kit (C0468) can be used.
4.Staining and Observation (Optional)
a.Dissolve dyes in the clearing reagent and immerse the cleared samples at room temperature for 30-60 minutes. For fluorescent dyes, stain in the dark.
Note 1: The Staining Solution can be prepared with water, clearing reagent, or other appropriate solvents, as needed.
Note 2: Adjust staining time based on sample type and dye properties, referring to relevant literature.
Note 3: Not all dyes are compatible with this product. Check their compatibility through experimentation.
Note 4: Regular staining can be performed either before or after clearing, depending on whether clearing affects staining intensity. Fluorescent staining should be performed after clearing because fluorescence intensity decreases over time.
b.Wash stained samples gently using the clearing reagent for 3-4 cycles over 30-60 minutes.
c.Place a drop of clearing reagent or pure water on a microscope slide, carefully position the sample using tweezers, and cover with a coverslip.
Note: Select an appropriate mounting medium based on experimental needs.
d.Examine conventional stained samples by a light microscope, or fluorescent samples by a fluorescence microscope.