There are numerous mounting media available for use with the Masson Trichrome kit (HT15) and other routine histology staining and special stain procedures. The names and catalog numbers are listed below:
- Orango/Limonene Mount: O8015
- DPX: 06522, 100579, 317616
- Entellan: 107960, 107961, and 100869
- Eukitt: 03989 and 05393
- Krystalon: 64969
推薦產品
診斷意義
可能有助于研究以纤维化和营养不良变化为特征的结缔组织和肌肉疾病。
原則
胶原和肌肉纤维先后采用Biebrich Scarlet-Acid Fuchsin、PTA/PMA和Aniline Blue处理后，会出现明显的染色差异。细胞质和肌肉纤维染色为红色，而胶原则显示为蓝色。
僅套裝組件
產品號碼
描述
- Aniline Blue Solution (kit only) 250 mL
- Biebrich Scarlet-Acid Fuchsin Solution (HT151) 250 mL
- Phosphomolybdic Acid Solution (HT153) 250 mL
- Phosphotungstic Acid Solution (HT152) 250 mL
相關產品
產品號碼
描述
訂價
訊號詞
Danger
危險聲明
危險分類
Aquatic Chronic 3 - Eye Dam. 1 - Ox. Liq. 3 - Skin Corr. 1B
儲存類別代碼
5.1B - Oxidizing hazardous materials
閃點（°F）
Not applicable
閃點（°C）
Not applicable
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分析證明 (COA)
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What type of mounting media should be used with the Masson Trichrome kit, HT15?
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Is it correct that hematoxylin is not included in this kit? Is there any concern in using hematoxylin solution, Mayer's (MHS16) instead of Weigert's iron hematoxylin solution?
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The HT15 kit does not include a hematoxylin solution. It is not possible to substitute Mayers Hematoxylin for Weigert's Iron Hematoxylin. Mayers hematoxylin is aluminum-based and if used, the nuclei staining tends to fade when exposed to the remaining reagents used in the Trichrome procedure. It is recommended to use Weigert's Iron Hematoxylin as it is an iron-based hematoxylin formulation that will not fade or be removed from the nuclei after exposure to the remaining staining reagents utilized in the staining procedure.
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Is it possible to use the Weigert's Iron Hematoxylin from Millipore Sigma with Trichrome reagents from other companies?
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The purpose of using Weigert's Iron Hematoxylin instead of a Mayer's, Gill's, or Harris hematoxylin is related to the dye being retained in the tissue. When tissue is stained with an aluminum-based hematoxylin, subsequent Trichrome reagents can cause the aluminum-based hematoxylin staining to leak out from the nucleus. However, if an iron-based hematoxylin, such as Weigert's Iron Hematoxylin, is used, the Trichrome reagents will not leach out the hematoxylin from the tissues stained with it.
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Could you please clarify if the correct dilution to use in the Masson's Trichrome kit protocol is 1% acetic acid, as there seems to be a discrepancy in the instructions? It has been observed that when using the true 1% acetic acid, the contrast is lower compared to the recommended ratio of about 17.5%. Is it okay to proceed using the 17.5% dilution instead of the true 1% acetic acid?
1 answer-
It is necessary to start with a 1N Acetic Acid solution for the dilution to 1%. If the glacial acetic acid is 100%, the amount used to produce 1 liter of a 1N acetic acid solution is 57.194 ml, which is the equivalent to 5.72%. By multiplying 5.72 by 17.5% (5.72 X 0.175), this comes out to be 1%. If an acid that is not 100% is used, suboptimal staining might occur. However, as long as the glacial acetic acid is within a percent or two of 100%, it should not be an issue. Either the procedure method or a direct dilution from glacial acetic acid should produce the same results. If an acid that is not 100% is used, suboptimal staining might occur.
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When using the kit for the procedure, is there a specific thickness recommended for sectioning tissue samples? Intended use of tissue samples consisting of formalin-fixed paraffin-embedded rat left ventricular tissue of 5 micrometers for IHC.
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It is recommended that specimen collection be carried out in accordance with CLSI document M29-A3. No known test method can offer complete assurance that blood samples or tissue will not transmit infection. Therefore, all blood derivatives or tissue specimens should be considered potentially infectious. Any well-fixed paraffin section cut at 5–6 microns may be used. Incorporate appropriate control slides. his applies to both the Gomori Trichrome Procedure and the Masson Trichrome procedures.
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What could be causing a tissue sample to stain well initially but then have the dye disappear?
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The staining disappeared after coverslipping, likely due to prolonged dehydration in alcohol. The use of alkaline fluorescent mounting media may have caused the dye to fade. Consider coverslipping with permanent mounting media, such as DPX. If using Neo Mount, clear in Neo Clear without xylene; if using DPX Mounting Media, clear the slides with xylene before coverslipping.
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How many slides can be stained with one kit? Can the reagents be reused?
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The number of slides that can be stained with the HT15 kit varies significantly. The Biebrich Scarlet-Acid Fuchsin and Aniline Blue solutions can typically be reused 4-5 times, or more. The combined working solution of products HT15-2 and HT15-3 is intended for one-time use only, and it's possible to stain anywhere from 1 to 12-14 slides per run. The kit is designed for approximately 25 uses, and the number of slides stained with the kit could range from 25 to perhaps 300 slides per kit. No specific claims are made regarding the number of stained slides, as this can vary depending on whether the slides are stained all at once or over a period of several weeks or months. In general, the working staining reagents are replaced when the slide quality starts to deteriorate.
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Is it possible to use item 1159730002 in place of HT1079 for the Weigert's iron hematoxylin solution?
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Yes, product number 1159730002 is certainly suitable for use with the HT15 kit. It is acceptable to use a Weigert's Iron Hematoxylin that is not mentioned in the HT15 procedure for the Masson Trichrome kit, HT15.
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How many tests can be performed with this kit? Masson Tri Chrome #HT15-1kt
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The number of slides that can be stained with the HT15 kit is highly variable. The Biebrich Scarlet-Acid Fuchsin and the Aniline Blue solutions can generally be reused at least 4-5 times. The limiting reagents in the kit are the acid differentiators – Phosphomolybdic and Phosphotungstic Acid Solutions. Typically, 10 ml of each reagent (combined volume of 20 mL) is combined with 20 mL water. This 40 ml of acid differentiator is intended for 1 time use. The number of slides to be stained in each run is variable. The kit was formulated to 25 staining runs. The number of slides stained with the kit could vary from 25 to perhaps 300 slides per kit. No specific claims are made for the number of stained slides.
Please see the link below to review the kit user guide:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/274/001/procedure-ht15ug-ms.pdfHelpful?
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