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CAS RN: | Product Number: T3530

Tissue-Clearing Reagent TOMEI [for Plants]


Purity:
Synonyms:
  • 2,2'-Thiodiethanol (contains Propyl Gallate, PBS and DMSO) [Tissue-Clearing Reagent for Plants]
Product Documents:
100ML
$105.00
1   3   17  

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Supplemental Product Information:


The transparency technique "TOMEI" for plant analysis was developed by Prof. Matsunaga et al. at Tokyo University of Science. This method enables the clearing of Oryza sativa, Arabidopsis thaliana and many others in just a few hours. Using this technique, deeper internal structure can be clearly observed. TCI offers TOMEI reagent (Product Number: T3530) for this transparency techique.

Please see a brochure (PDF) of how to use.

Product Number T3530
Physical State (20 deg.C) Liquid
Storage Temperature Room Temperature (Recommended in a cool and dark place, <15°C)
Condition to Avoid Light Sensitive
Specifications
Appearance Colorless to Light yellow clear liquid
Functionality test(transparency) to pass test
Properties (reference)
GHS
Pictogram Pictogram
Signal Word Warning
Hazard Statements H315 : Causes skin irritation.
H319 : Causes serious eye irritation.
Precautionary Statements P264 : Wash skin thoroughly after handling.
P280 : Wear protective gloves/ eye protection/ face protection.
P337 + P313 : If eye irritation persists: Get medical advice/ attention.
P305 + P351 + P338 : IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing.
P302 + P352 : IF ON SKIN: Wash with plenty of soap and water.
P332 + P313 : If skin irritation occurs: Get medical advice/ attention.
P362 : Take off contaminated clothing and wash before reuse.
Related Laws:
Transport Information:
HS Number 2930.90.9251
Application
Clearing of fungi-infected plants

References


Application
TOMEI-I, Technique for analysis with only fluorescent dye

The transparency technique “TOMEI-I” is adequate for observation with only fluorescent dyes such as DAPI staining, Calcofluor White staining, etc.

【Reagents】
・Fixative solution (Acetic acid : Ethanol = 1 : 3) *Preparation at time of use is recommended.
・PBS
・70% Ethanol (diluted with PBS)
・30% Ethanol (diluted with PBS)
・Staining solution
・Transparency reagent TOMEI (TCI product code T3530)
*For root specimens, 20% TOMEI diluted with PBS is necessary.

【Procedure】
The transparency method of the final process differs depending on the above ground part and the underground part (root).

< Fixing >
1) Fix the sample with adequate amount of fixative solution at room temperature.
(Fixation time is determined by the sample size and plant type.)
(In case of a seedling of Arabidopsis thaliana, it usually takes for about 1 to 2 hours.)
2) Remove fixation solution and add 70% ethanol, then let it rest for 5min at room temperature.
3) Remove 70% ethanol and add 30% ethanol, then let it rest for 5min at room temperature.
4) Remove 30% ethanol PBS and add PBS, then let it rest for 5min at room temperature.

< Staining >
5) Remove PBS and add the staining solution, then let it rest with shading at room temperature. *1
(In case of double staining, re-stain after washing.)

< Washing >
6) Remove the staining solution and add PBS, then let it rest for 10min at room temperature.
7) Remove PBS and add PBS, then let it rest for 10min at room temperature.
8) Remove PBS and add PBS, then let it rest for 10min at room temperature.

< Clearing >
* For above ground part
9) Remove PBS and add TOMEI, then let it rest with shading at room temperature for 20min.
(The time to treat should be considered depending on sample.)
10) Mount the sample on a microscope slide with TOMEI and observe it.
*For underground part (root)
9) Remove PBS and add 20% TOMEI in PBS, then let it rest with shading at room temperature for 10min.
10) Mount the sample on a microscope slide with 20% TOMEI in PBS and observe it.

*1 : The treatment time of DAPI staining is for 30min with 5 μg/mL and the treatment time of Calcofluor White staining is for 10min with 1 g/L of Calcofluor White M3R and 0.5 g/L of Evans Blue, but their adjustments are needed for the purpose.

References


Application
TOMEI-II, Technique for detection of fluorescent protein

The transparency technique “TOMEI-II” is adequate for observation fluorescent protein such as GFP or YFP co-expressed as a reporter gene.

【Reagents】
・Fixative solution (4% paraformaldehyde in PBS)
 *Preparation at time of use is recommended.
・PBS
・Staining solution
・Transparency reagent TOMEI (TCI product code T3530)
・10%, 30%, 50% , 70% TOMEI (diluted with PBS)
 *For the root specimens, 20% TOMEI diluted with PBS is necessary.

【Procedure】
The transparency method of the final process differs depending on the above ground part and the underground part (root).

< Fixing >
1) Fix the sample with adequate amount of fixative solution at room temperature.
 (Fixation time is determined by the sample size and plant type.)
 (Deaerate in fixative solution using vacuum pump or syringe when the sample is above ground part.)
2) Remove fixation solution and add PBS, then let it rest for 5min at room temperature.
3) Remove PBS and add PBS again, then let it rest for 10min at room temperature.
4) Remove PBS and add PBS again, then let it rest for 10min at room temperature.

< Staining >
5) Remove PBS and add the staining solution, then let it rest with shading at room temperature.*1
 (In case of double staining, re-stain after washing.)

< Washing >
6) Remove the staining solution and add PBS, then let it rest for 10min at room temperature.
7) Remove PBS and add PBS, then let it rest for 10min at room temperature.
8) Remove PBS and add PBS, then let it rest for 10min at room temperature

< Clearing >
*For above ground part
9) Remove PBS and add 10% TOMEI in PBS, then let it rest with shading at room temperature for 10min.
10) Remove 10% PBS and add 30% TOMEI in PBS, then let it rest with shading at room temperature for 10min.
11) Remove 30% TOMEI in PBS and add 50% TOMEI in PBS, then let it rest with shading at room temperature for 10min.
12) Remove 50% TOMEI in PBS and add 70% TOMEI in PBS, then let it rest with shading at room temperature for 10min.
13) Remove 70% TOMEI in PBS and add 100% TOMEI in PBS, then let it rest with shading at room temperature for 10min.
14) Remove 100% TOMEI in PBS and add 100% TOMEI, then let it rest with shading at room temperature for 1hour.
 (The time to treat should be considered depending on sample.)
15) Mount the sample on a microscope slide with TOMEI and observe it. *2
*For underground part (root)
9) Remove PBS and add 20% TOMEI in PBS, then let it rest with shading at room temperature for 10min.
10) Mount the sample on a microscope slide with 20% TOMEI in PBS and observe it.

*1 : The treatment time of DAPI staining is for 30min with 5 µg/mL and the treatment time of Calcofluor White staining is for 10min with 1 g/L of Calcofluor White M3R and 0.5 g/L of Evans Blue, but their adjustments are needed for the purpose. *2 : When the fluorescence of fluorescent protein reduces, after the treatment of 50% TOMEI in PBS, it may be possible to be solved by being mounted with 60% TOMEI in PBS after the treatment of 60% TOMEI in PBS for 1hour.

References


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