Principles, Method Types, and Application Selection of Common Trichrome Staining Systems
Principles, Method Types, and Application Selection of Common Trichrome Staining Systems
Trichrome staining refers to a group of histochemical methods that use nuclear staining, cytoplasmic/muscle fiber staining, collagen or matrix counterstaining, and differentiation steps to display multiple structural components in the same tissue section. Its core value does not lie simply in producing three colors, but in establishing staining systems for morphological interpretation, fibrosis evaluation, bone matrix observation, and tissue structural stratification through the differential responses of tissue components to dyes, mordants, and differentiation conditions.
Keywords: trichrome staining; Masson staining; Goldner staining; Mallory staining; Azan staining; Gomori staining; Pollak staining; Van Gieson staining; collagen fibers; bone matrix; fibrosis; histochemical staining
1、Basic Logic of Trichrome Staining Systems
1.1 Structural Targets
(1) Cell nuclei
Trichrome staining usually first visualizes cell nuclei to provide a basis for tissue structural localization. Common nuclear stains include Weigert iron hematoxylin, Heidenhain iron hematoxylin, and some modified hematoxylin systems. Since subsequent steps often involve acidic staining solutions, phosphomolybdic acid/phosphotungstic acid differentiation, and collagen counterstaining, acid-resistant iron hematoxylin is more suitable for multistep trichrome staining procedures and can reduce nuclear fading during subsequent acid treatment.
(2) Cytoplasm and muscle fibers
Cytoplasm, muscle fibers, red blood cells, and some intracellular protein structures are commonly stained with acid fuchsin, ponceau, Biebrich Scarlet, Orange G, and similar dyes, usually appearing red, pink, or orange-red. This step determines the basic contrast between cellular components and collagenous matrix and is critical for clearly distinguishing muscle fibers from collagen in trichrome staining.
(3) Collagen and extracellular matrix
Collagen fibers, connective tissue matrix, bone matrix, and fibrotic areas are often counterstained with acidic dyes such as aniline blue, Light Green SF, or fast green. Different counterstains determine whether collagen or matrix finally appears blue, green, or blue-green, and also affect subsequent image analysis, fibrosis quantification, and tissue structure interpretation.
1.2 Key Points of Staining Mechanism
(1) Dye competition
Trichrome staining depends on competitive binding of different acidic dyes to tissue proteins. The first red dye binds initially to cytoplasm, muscle fibers, and some collagen areas. Then, through differentiation/mordanting steps using phosphomolybdic acid or phosphotungstic acid, collagen areas become more receptive to the final counterstain. This process determines the degree of color separation between collagen and cellular components.
(2) Differentiation control
Insufficient differentiation leaves residual red dye and makes collagen difficult to distinguish. Excessive differentiation may fade cellular structures and reduce tissue layering. The stability of trichrome staining largely depends on differentiation time, acidic conditions, washing intensity, fixation conditions, and section thickness.
(3) Counterstain selection
The counterstain determines the final color of collagen and matrix. Aniline blue is commonly used for blue collagen staining, Light Green SF is commonly used for green collagen or bone matrix staining, and fast green is often used in fast green Masson staining, modified trichrome staining, and osteochondral counterstaining systems. Interpretation standards from different counterstaining systems should not be directly mixed.
Table 1 Structural Roles and Common Dyes in Trichrome Staining
Staining step | Main target structure | Common dyes or reagents | Typical color | Methodological significance |
Nuclear staining | Cell nuclei, nuclear chromatin | Weigert iron hematoxylin, Heidenhain iron hematoxylin | Blue-black, dark blue | Provides tissue and cellular localization |
Cytoplasm/muscle fiber staining | Cytoplasm, muscle fibers, red blood cells | Acid fuchsin, ponceau, Biebrich Scarlet, Orange G | Red, orange-red | Distinguishes cellular components from matrix |
Differentiation/mordanting | Dye displacement in collagen regions | Phosphomolybdic acid, phosphotungstic acid, acetic acid systems | No independent color | Determines collagen counterstain selectivity |
Collagen/matrix counterstaining | Collagen fibers, bone matrix, extracellular matrix | Aniline blue, Light Green SF, fast green | Blue, green, blue-green | Shows collagen deposition and matrix distribution |
Dehydration, clearing, mounting | Post-staining slide preservation | Ethanol, xylene, neutral resin | No independent color | Ensures long-term observation and image acquisition |
2、Major Trichrome Staining Systems
2.1 Masson Trichrome Staining
(1) Method positioning
Masson trichrome staining is one of the most widely used trichrome staining systems for observing collagen fibers and fibrosis. It is mainly used to distinguish cell nuclei, cytoplasm/muscle fibers, and collagen fibers. Its applications include evaluation of liver fibrosis, renal interstitial fibrosis, pulmonary fibrosis, myocardial fibrosis, skin scarring, tumor stromal fibrosis, and tissue injury repair.
(2) Basic principle
Masson trichrome staining stratifies tissue components based on differences in the binding ability of acidic dyes to different tissue proteins. Cell nuclei are first stained with iron hematoxylin; cytoplasm and muscle fibers are stained red with acid fuchsin, ponceau, or Biebrich Scarlet-type dyes; then phosphomolybdic acid or phosphotungstic acid differentiation allows collagen regions to accept final counterstains such as aniline blue, Light Green SF, or fast green. Differentiation is the core step of Masson staining and determines the clarity of the boundary between red cellular components and blue or green collagen.
(3) Typical results
Typical results include blue-black nuclei, red cytoplasm, muscle fibers and red blood cells, and blue or green collagen fibers. Aniline blue Masson staining shows collagen in blue and is suitable for fibrosis area analysis. Light Green SF or fast green Masson staining shows collagen or matrix in green and is suitable for green collagen output, microscopic image presentation, and method comparison.
(4) Application focus
Masson trichrome staining focuses on the extent of collagen deposition, fibrosis distribution, the boundary between muscle fibers and interstitium, and structural changes in tissue repair areas. In liver fibrosis, attention should be paid to portal areas, bridging fibrosis in portal regions, and pseudolobule formation. In myocardial tissue, perivascular collagen, interstitial myocardial collagen, and scar areas should be evaluated. In tumor stroma, interpretation should combine tumor cell nests, stromal proportion, and collagen bundle arrangement.
(5) Limitations
Masson staining displays collagen/matrix regions in a histochemical sense and does not distinguish collagen subtypes. If type I collagen, type III collagen, or collagen maturation needs to be differentiated, immunohistochemistry, polarized Sirius Red staining, or molecular assays should be combined. For image quantification, section thickness, staining batch, scanning parameters, and color thresholds should be standardized.
2.2 Goldner Trichrome Staining
(1) Method positioning
Goldner trichrome staining is commonly used for bone tissue, mineralized tissue, and bone formation-related samples. It is suitable for observing spatial relationships among bone matrix, osteoid, cellular components, and connective tissue matrix. This method has high application value in bone metabolism, bone repair, bone implant material evaluation, bone formation models, and mineralized matrix studies.
(2) Basic principle
The Goldner system establishes structural stratification in bone tissue through nuclear staining, acidic red staining, and green counterstaining. Nuclei usually appear dark; cellular components and some unmineralized areas appear red or orange-red; mature bone matrix, mineralized matrix, or collagenous areas mostly appear green. Light Green SF is a representative green counterstain in this system and highlights bone matrix and mature collagenous regions.
(3) Typical results
In bone tissue, mineralized bone matrix is often green, osteoid or cellular areas may appear red to orange-red, and nuclei appear dark. This system is useful for observing trabecular bone structure, bone matrix deposition, bone formation boundaries, bone repair processes, and distribution of newly formed bone around materials.
(4) Application focus
The focus of Goldner staining is not simply measuring collagen area, but distinguishing mineralized bone matrix, unmineralized osteoid, cellular components, and bone tissue background. For bone formation models, bone defect repair, and tissue reaction around bone implants, the Goldner system better reflects bone tissue stratification and mineralization status than ordinary Masson staining.
(5) Limitations
Decalcification method, decalcification time, and embedding method significantly affect bone matrix staining. Excessive decalcification may reduce matrix preservation and color stability, while undecalcified samples place higher demands on section quality and staining penetration. In comparative bone tissue studies, decalcification, sectioning, and staining conditions should remain consistent within the same experiment.
2.3 Mallory Trichrome Staining
(1) Method positioning
Mallory trichrome staining is a classic histochemical trichrome system used to display collagen fibers, connective tissue, muscle fibers, red blood cells, and some cellular components in multiple color layers. It is suitable for connective tissue structure presentation, multicomponent tissue comparison, and methodological comparison of trichrome staining.
(2) Basic principle
The Mallory system usually establishes color stratification through combinations of dyes such as acid fuchsin, Orange G, and aniline blue. Different tissue components show blue, red, or orange colors because of differences in protein properties, tissue density, and dye affinity. Aniline blue is mainly used for blue staining of collagen and connective tissue; Orange G is commonly used to contrast red blood cells or keratinized structures; acid fuchsin-type dyes stain cytoplasm and muscle fibers.
(3) Typical results
Collagen and connective tissue are usually blue; cytoplasm or muscle fibers may appear red to red-orange; red blood cells or keratinized structures may appear orange. This method provides rich color layering and is suitable for observing tissue architecture, collagen bundle distribution, and relationships between cellular components and interstitium.
(4) Relationship with PTAH systems
Mallory phosphotungstic acid hematoxylin staining (PTAH) is related to Mallory methods, but its application focus is more oriented toward muscle fibers, striations, fibrin, and partial tissue structure contrast. It is not equivalent to a typical trichrome system centered on collagen counterstaining. It may be included in the text as a supplementary Mallory-related histochemical method, but should not be confused with collagen counterstaining systems such as Masson or Goldner.
(5) Limitations
The Mallory system has many operational steps, and differentiation conditions significantly affect color layering. If used for collagen area quantification, it is necessary to confirm that blue structures correspond to the target collagen regions. If the goal is standardized fibrosis area analysis, Masson systems are usually easier for establishing stable thresholds.
2.4 Azan Trichrome Staining
(1) Method positioning
Azan trichrome staining is a trichrome system that uses dyes such as azocarmine, Orange G, and aniline blue to construct multicolor contrast. It is often used for structural stratification of collagen, connective tissue, muscle tissue, and cellular components. The term “Azan” usually refers to azocarmine- and aniline blue-related combinations, emphasizing differential display of multicomponent tissue structures.
(2) Basic principle
The Azan system uses azocarmine-type dyes to display nuclei and some cytoplasmic structures, Orange G to provide orange contrast for red blood cells or certain cytoplasmic components, and aniline blue to show collagen and connective tissue. The key lies in coordinating multidye competition and differentiation steps so that different tissue components exhibit stable stratification.
(3) Typical results
Nuclei and some cytoplasmic structures may appear red to reddish-purple; red blood cells or specific cytoplasmic components may appear orange; collagen and connective tissue are mostly blue. Compared with Masson staining, Azan emphasizes overall tissue structural layering rather than only fibrosis area evaluation.
(4) Application focus
Azan is suitable for observing collagen bundle distribution, connective tissue architecture, cell-matrix relationships, and changes in tissue organization. It provides rich structural information for embryonic tissues, connective tissue-rich samples, glandular interstitium, and samples requiring multicomponent stratified display.
(5) Limitations
Azan staining is sensitive to differentiation conditions, and results may vary among laboratories. If the blue collagen staining system is changed to green counterstaining, it should be treated as a methodological modification and should not be considered directly equivalent to classical Azan results.
2.5 Gomori Trichrome Staining
(1) Method positioning
Gomori trichrome staining is commonly used in muscle pathology, connective tissue observation, and tissue structural stratification. It is particularly suitable for observing muscle fibers, connective tissue, and abnormal mitochondria-related structures in skeletal muscle pathology. A representative application is modified Gomori trichrome staining for abnormal structure detection in muscle biopsies.
(2) Basic principle
Gomori trichrome staining displays muscle fibers, connective tissue, and abnormal structures through nuclear staining, complex acidic dye staining, and differentiation steps. Different modified versions have substantial differences in formulation, and color output is not completely consistent. Therefore, interpretation must be based on the specific method system used.
(3) Typical results
Muscle fibers, collagen/interstitium, and some abnormal structures may show different color layers. Modified Gomori trichrome staining is often used in skeletal muscle biopsies to observe muscle fiber structural abnormalities, ragged red fiber-like changes, interstitial proliferation, and pathological deposits.
(4) Application focus
Gomori trichrome is more oriented toward muscle pathology and specific abnormal structure display. It is not the preferred method for simple collagen area evaluation. If the research goal is quantification of liver, lung, kidney, or myocardial fibrosis, Masson aniline blue or Light Green methods are usually more direct.
(5) Limitations
There are many modified versions of Gomori systems, and color expression may vary among kits or laboratory formulations. For cross-batch comparison, same-batch positive controls should be included and interpretation criteria should be standardized.
2.6 Pollak Trichrome Staining
(1) Method positioning
Pollak trichrome staining is a method for stratified display of multiple tissue structures and can be used to contrast connective tissue, cellular components, and matrix structures. Compared with Masson and Goldner systems, Pollak is less frequently used in routine fibrosis quantification and bone matrix interpretation, but it can serve as an expanded method within trichrome staining.
(2) Basic principle
Pollak trichrome staining usually relies on multiple acidic dyes and differentiation steps to build tissue structural contrast. Its focus is not on displaying collagen alone, but on presenting cellular components, interstitium, and tissue background through multicolor contrast.
(3) Typical results
Specific color expression depends on the formulation. Some systems can display multicolor stratification among cellular components, collagenous matrix, and background tissue and are used for tissue structure observation and methodological comparison.
(4) Application focus
Pollak trichrome staining is more suitable as a multistructural tissue component stratification method for comparison with Masson, Mallory, Azan, and other systems. If the research focus is fibrosis quantification, a more standardized Masson system should still be prioritized.
(5) Limitations
Whether the Pollak system directly uses green or blue matrix counterstaining must be judged according to the specific formulation. If formulations differ, it should be handled as a method-type trichrome staining product rather than being directly assigned to a specific counterstain dye system.
2.7 Van Gieson-Related Staining
(1) Method positioning
Van Gieson staining is commonly used to distinguish collagen from muscular tissue. Typical results show collagen in red and muscle fibers/cytoplasm in yellow. Strictly speaking, it is not a typical trichrome system built around the complete sequence of “nuclear staining—cytoplasm/muscle fiber staining—collagen counterstaining,” but it is often compared with Masson, Goldner, and other methods in connective tissue staining.
(2) Basic principle
The Van Gieson system uses components such as acid fuchsin and picric acid to give collagen and muscular tissue different colors. Collagen is usually red, muscle fibers and cytoplasm are yellow, and nuclei are shown by the corresponding nuclear stain.
(3) Application focus
Van Gieson is suitable for rapid display of differences between collagen fibers and muscular tissue and can be used as a collagen observation control method outside trichrome staining. It has value in rapid tissue structure comparison, preliminary observation of collagen distribution, and methodological referencing.
(4) Limitations
Van Gieson red collagen staining differs from Masson blue/green collagen display and should not be directly combined with Masson or Goldner results for quantification. For image analysis, independent color thresholds and interpretation criteria should be established.
Table 2 Comparison of Common Trichrome and Related Staining Systems
Staining system | Core use | Typical collagen/matrix color | Representative dye direction | Preferred application scenarios |
Masson trichrome | Collagen, fibrosis, muscle fiber differentiation | Blue or green | Aniline blue, Light Green SF, fast green | Liver, lung, kidney, myocardium, skin fibrosis |
Goldner trichrome | Bone matrix, mineralized tissue, bone formation | Green | Light Green SF as representative | Bone tissue, mineralized tissue, bone formation models |
Mallory trichrome | Stratification of collagen, connective tissue, and cellular components | Blue | Aniline blue as representative | Connective tissue structure, multicolor tissue presentation |
Azan trichrome | Multicolor stratification of collagen, cytoplasm, and red blood cells | Blue | Aniline blue as representative | Collagen bundles, stromal structure, tissue layering |
Gomori trichrome | Muscle pathology and tissue structural stratification | Depends on formulation | Should be judged by specific system | Skeletal muscle pathology, connective tissue observation |
Pollak trichrome | Stratification of multiple structural tissue components | Depends on formulation | Should be judged by specific system | Tissue structural stratification and methodological expansion |
Van Gieson-related staining | Distinction between collagen and muscular tissue | Red | Not centered on trichrome counterstain dyes | Rapid connective tissue comparison and control observation |
3、Counterstain Colors and Structural Interpretation
3.1 Blue Collagen Systems
(1) Method characteristics
Blue collagen systems are represented by aniline blue and are common in Masson, Mallory, and Azan-related staining. Collagen fibers, fibrotic areas, and connective tissue matrix are mostly blue to blue-purple, forming clear contrast with red cytoplasm, muscle fibers, and red blood cells.
(2) Application scenarios
Blue collagen systems are suitable for evaluating fibrosis in liver, kidney, lung, myocardium, skin, and tumor stroma. They are also suitable for observing collagen bundles, scar tissue, and connective tissue proliferation. For image analysis, blue collagen usually facilitates relatively stable color threshold establishment.
(3) Technical risks
Insufficient aniline blue staining may result in weak blue collagen staining. Excessive staining or insufficient washing may cause a blue background. For quantitative analysis, section thickness, staining batch, scanning parameters, and threshold settings should be kept consistent.
3.2 Green Collagen or Matrix Systems
(1) Method characteristics
Green counterstaining systems often involve Light Green SF or fast green. Light Green SF is more commonly used in Goldner trichrome, green Masson variants, and bone matrix display. Fast green is more common in fast green Masson staining, modified trichrome staining, and osteochondral counterstaining systems.
(2) Application scenarios
Green collagen or matrix systems are suitable for bone tissue, mineralized matrix, bone repair areas, and tissue presentations requiring green collagen output. The green signal in Goldner systems is more oriented toward bone matrix and mature collagenous regions, while fast green-related systems require interpretation according to the specific method.
(3) Technical risks
Overstaining with green dyes can cause a green background, blurring interstitial boundaries or overestimating positive areas. Green signals from fast green in different systems are not necessarily equivalent to collagen positivity, especially in cartilage, bone tissue, and counterstaining systems, where method background must be considered.
3.3 Red Collagen Control Systems
(1) Method characteristics
In Van Gieson-related staining, collagen is usually red, while muscle fibers and cytoplasm are yellow. These results differ from blue/green collagen systems in Masson or Goldner staining and are more suitable as control methods for collagen observation.
(2) Application scenarios
This system is suitable for rapid comparison of collagen and muscular tissue and can also be used alongside Masson, Goldner, and other trichrome systems for methodological comparison. However, collagen areas from different color systems should not be directly combined for calculation.
Table 3 Interpretive Differences Among Counterstain Colors
Counterstain type | Common systems | Typical color | Main applications | Interpretation notes |
Blue collagen system | Masson, Mallory, Azan | Blue, blue-purple | Collagen fibers, fibrosis, connective tissue stratification | Blue background may expand the positive area |
Green collagen/matrix system | Goldner, green Masson, fast green modified systems | Green, yellow-green, blue-green | Bone matrix, mineralized tissue, green collagen display | Green signal must be interpreted according to the specific system |
Red collagen control system | Van Gieson-related staining | Red | Rapid distinction between collagen and muscular tissue | Should not be directly combined with blue/green collagen systems for quantification |
Muscle pathology multicolor system | Gomori trichrome | Depends on formulation | Observation of muscle fibers, connective tissue, and abnormal structures | Not preferred for simple collagen area evaluation |
Osteochondral counterstaining system | Safranin O-fast green, etc. | Red, green, etc. | Contrast between cartilage matrix and background matrix | Fast green signal is not equivalent to trichrome collagen counterstaining |
4、Interpretation of Trichrome Staining Results
4.1 Collagen and Fibrosis
(1) Extent of collagen deposition
Masson trichrome staining is often used to evaluate the extent of collagen deposition. Collagen may be blue or green, depending on the counterstaining system. Interpretation should focus on whether collagen is located in the interstitium, around blood vessels, in capsules, in scar regions, or within tissue parenchyma.
(2) Degree of fibrosis
Fibrosis evaluation should not rely only on color intensity. It should combine positive area, structural distribution, degree of tissue destruction, and localization of lesion regions. Image analysis should standardize thresholds, section thickness, and staining batch.
(3) Background interference
Excessive counterstaining, insufficient differentiation, or inadequate washing can enhance background staining and overestimate collagen areas. Before interpretation, it should be confirmed that red cytoplasm/muscle fibers, nuclear staining, and collagen counterstaining are balanced.
4.2 Boundary Between Muscle Fibers and Collagen
(1) Masson system
Masson staining clearly distinguishes red muscle fibers from blue/green collagen and is suitable for observing interstitial fibrosis in myocardium, skeletal muscle, and smooth muscle tissue. If red staining is insufficient, muscle fiber boundaries become weak. If collagen counterstaining is excessive, cellular structures may be obscured.
(2) Gomori system
Gomori trichrome is more oriented toward muscle pathology and muscle fiber structural abnormality observation. It is suitable for overall interpretation of muscle fiber structure, interstitium, and specific abnormal structures. This system should not directly replace Masson staining for fibrosis area quantification.
(3) Mallory and Azan systems
Mallory and Azan emphasize multistructural stratification and are suitable for observing relationships among collagen, red blood cells, cytoplasm, and connective tissue. When used for fibrosis area quantification, color thresholds and definitions of positive structures should be confirmed.
4.3 Bone Matrix and Osteochondral Interface
(1) Goldner system
Goldner trichrome is suitable for displaying bone matrix and mineralized tissue structures. Light Green SF is often used to counterstain mature bone matrix or collagenous matrix. This system should be prioritized for bone formation models and bone tissue repair evaluation.
(2) Fast green-related counterstaining
In Safranin O-fast green, bone tissue Safranin-fast green, and cartilage-related staining, fast green is commonly used to contrast background matrix or bone tissue. These systems are related to typical trichrome staining, but their interpretation targets differ and should be distinguished from Masson or Goldner staining.
Table 4 Key Points for Interpreting Trichrome Staining Results
Interpretation target | Recommended system | Typical positive color | Interpretation focus | Main risk |
Liver, kidney, lung fibrosis | Masson aniline blue method | Blue collagen | Extent of collagen deposition and fibrosis area | Blue background may expand positive area |
Myocardial interstitial fibrosis | Masson aniline blue method or Light Green method | Blue or green collagen | Boundary between muscle fibers and interstitial collagen | Insufficient red staining affects boundary interpretation |
Bone matrix/mineralized tissue | Goldner trichrome | Green bone matrix | Trabeculae, mineralized matrix, and osteoid stratification | Excessive decalcification affects matrix staining |
Connective tissue structure presentation | Mallory or Azan | Blue collagen | Multilayer display of collagen, cytoplasm, and red blood cells | Improper differentiation causes color layer confusion |
Osteochondral interface | Fast green-related counterstaining or modified trichrome | Green/blue-green matrix | Contrast among cartilage, bone matrix, and background | Green is not necessarily equivalent to collagen positivity |
Muscle pathology | Gomori trichrome | Depends on formulation | Muscle fibers, interstitium, and abnormal structures | Not preferred for simple collagen quantification |
5、Trichrome Staining-Related Reagent Systems
5.1 Finished Method Systems and Tissue Compatibility
Table 5 Finished Trichrome Staining Method Systems and Tissue Compatibility
Cat. No. | Product Name | Grade/Specification | Corresponding staining system | Suitable samples | Main targets | Application positioning |
Goldner Tricolor Staining Solution | BioReagent, Biological Stain, for microscopy | Goldner trichrome staining | Bone tissue, mineralized tissue, bone repair samples | Bone matrix, mineralized matrix, collagenous matrix | Suitable for bone tissue structural stratification and closely related to green counterstaining | |
Masson's Trichrome Staining Kit | BioReagent, Biological Stain, for microscopy | Masson trichrome staining | Liver, lung, kidney, myocardium, skin, tumor tissue | Collagen fibers, muscle fibers, cytoplasmic structures | Common system for fibrosis evaluation and collagen deposition observation | |
Masson Trichrome Staining Kit (Fast Green Method) | BioReagent, Biological Stain, for microscopy | Masson trichrome staining; fast green method | Fibrotic tissue, connective tissue, matrix-rich samples | Green collagen/matrix, muscle fibers, nuclei | Used for green collagen/matrix display and suitable for fast green counterstaining systems | |
Modified Masson Trichrome Staining Solution | BioReagent, Biological Stain, for microscopy | Modified Masson trichrome staining | Tissue sections, fibrosis models, connective tissue samples | Collagen, muscle fibers, cytoplasmic structures | Used for contrast between collagen and cellular components; can serve as a modified Masson system | |
Pollak Trichrome Staining Solution | BioReagent, Biological Stain, for microscopy | Pollak trichrome staining | Samples requiring tissue structural stratification | Cellular components, interstitium, tissue background | Used for multistructural tissue stratification and can serve as an expanded trichrome method | |
Mallory PTAH Stain Solution (Spontaneous Oxidation Method) | BioReagent,Biological dye grade,for microscopy | Mallory PTAH-related system | Muscle tissue, fibrin-related samples | Muscle fibers, fibrin, tissue structure | Used for contrast of muscle fibers, fibrin, and tissue structure; suitable as a supplementary Mallory-related histochemical system | |
Mallory Phosphotungstic Acid Hematoxylin Staining Solution (PTAH Natural Oxidation Method) | BioReagent,Biological Stain,for microscopy | Mallory PTAH-related system | Muscle tissue, connective tissue, tissue structure samples | Muscle fibers, fibrous components, tissue structure | Used for tissue structure, muscle fibers, and fibrous component contrast | |
Mallory's Phosphotungstic Acid Hematoxylin Staining Kit (PTAH Chemical Oxidation) | BioReagent, Biological Stain, for microscopy | Mallory PTAH-related system | Muscle tissue, fibrin-related samples | Muscle fibers, striations, fibrin | Can serve as a Mallory-related histochemical product and supplement trichrome staining system background | |
Van Gieson Staining Solution | BioReagent, Biological Stain, for microscopy | Van Gieson-related staining | Connective tissue, muscle tissue, skin, vascular tissue | Collagen, muscular tissue | Used to distinguish collagen from muscular tissue; can serve as a control method for Masson, Goldner, and other trichrome systems | |
Van Gieson Staining Kit | BioReagent, Biological Stain, for microscopy | Van Gieson-related staining | Connective tissue, muscle tissue, fibrotic samples | Red-stained collagen, yellow muscular tissue | Used for red collagen staining and muscular tissue differentiation; suitable for comparison with trichrome staining results | |
Modified Van Gieson Staining Solution | BioReagent, Biological Stain, for microscopy | Modified Van Gieson staining | Connective tissue, collagen-rich samples | Collagen, tissue background | Used for contrast between collagen and tissue background; can supplement collagen staining outside trichrome systems |
5.2 Collagen and Matrix Counterstaining Solutions
Table 6 Collagen and Matrix Counterstaining Solution Configuration
Cat. No. | Product Name | Grade/Specification | Corresponding dye direction | Applicable system | Main targets | Application positioning |
Light Green SF Yellowish Stain Solution (1%) | BioReagent,Biological Stain,for microscopy,1% | Light Green SF | Goldner trichrome, green Masson variants | Collagen, bone matrix, green extracellular matrix | Used for green collagen/bone matrix display; key product for green counterstaining | |
Bright green staining solution (1%) | BioReagent,Biological Stain,for microscopy,1% | Brilliant green-related | Modified green counterstaining systems | Collagen or matrix background | Can be used as supporting information for green collagen or matrix display systems | |
Brilliant Green Staining Solution (2%) | BioReagent,Biological Stain,for microscopy,2% | Brilliant green-related | Modified green counterstaining systems | Collagen or matrix background | Suitable for method condition comparison in green counterstaining systems; use must be judged with specific formulation | |
Aniline Blue Stain Solution (0.1%) | BioReagent,Biological Stain,for microscopy,0.1% | Aniline blue | Masson, Mallory, Azan-related systems | Collagen fibers, connective tissue | Used for low-concentration aniline blue condition comparison and collagen blue staining optimization | |
Aniline Blue Stain Solution (0.5%) | BioReagent,Biological Stain,for microscopy,0.5% | Aniline blue | Masson, Mallory, Azan-related systems | Collagen fibers, fibrotic matrix | Suitable for collagen blue staining in trichrome systems | |
Aniline Blue Stain Solution (1%) | BioReagent,Biological Stain,for microscopy,1% | Aniline blue | Masson, Mallory, Azan-related systems | Collagen fibers, connective tissue matrix | Used for blue staining of collagen fibers in trichrome staining; core product of the aniline blue system | |
Aniline Blue Stain Solution (2%) | 2% | Aniline blue | High-concentration collagen blue staining systems | Collagen, connective tissue | Suitable for higher-concentration aniline blue staining systems or method condition comparison | |
Aniline blue staining solution (5%) | BioReagent,Biological Stain,for microscopy,5% | Aniline blue | High-concentration collagen blue staining systems | Collagen, matrix structures | Suitable for high-concentration aniline blue systems or staining solution preparation comparison | |
Fast Green FCF Staining Solution (0.1%) | BioReagent,Biological Stain,for microscopy,0.1% | Fast green | Fast green Masson, modified trichrome, matrix counterstaining | Green matrix, background structures | Used for fast green-related green matrix display; suitable for modified trichrome or matrix counterstaining discussion | |
Fast Green FCF Staining Solution (0.5%) | BioReagent,Biological Stain,for microscopy,0.5% | Fast green | Fast green Masson, osteochondral matrix counterstaining | Collagen/matrix, osteochondral background | Used for fast green Masson, osteochondral matrix counterstaining, or green matrix staining condition comparison | |
Fast Green FCF Ethanol Staining Solution (0.5%) | BioReagent,Biological Stain,for microscopy,0.5% | Fast green | Alcoholic fast green systems, osteochondral counterstaining | Green matrix, background tissue | Suitable for alcoholic fast green systems; can be used for osteochondral or modified trichrome-related counterstaining | |
Acid Fast Green Staining Solution (0.1%) | BioReagent,Biological Stain,for microscopy,0.1% | Fast green | Acidic counterstaining systems, modified trichrome | Green collagen/matrix | Closer to acidic counterstaining logic in trichrome systems; suitable for inclusion as a fast green counterstaining product | |
Basic Fast Green Staining Solution | BioReagent,Biological Stain,for microscopy | Fast green-related | Non-typical trichrome counterstaining systems | Depends on method | Not fully equivalent to acid fast green in typical trichrome systems; can supplement fast green-related products |
5.3 Supporting Reagents for Nuclear Staining and Cellular Component Visualization
Table 7 Supporting Reagents for Nuclear Staining and Cellular Component Visualization
Cat. No. | Product Name | Grade/Specification | Corresponding staining system | Suitable samples | Main targets |
Lichun Red Acid Fuchsin Staining Solution | BioReagent,Biological Stain,for microscopy | Masson trichrome, modified Masson system | Fibrotic tissue, muscle tissue, routine tissue sections | Cytoplasm, muscle fibers, red blood cells | |
Ponceau S | 10x | Masson-related system | Tissue sections, protein structure display samples | Cellular structures, muscle fibers | |
Orange G6 Staining Solution | BioReagent,Biological Stain,for microscopy | Mallory trichrome, Azan-related systems | Connective tissue, keratinized epithelial tissue, samples containing red blood cells | Red blood cells, keratinized materials, some cytoplasmic structures | |
Weigert's Hematoxylin Staining Kit | BioReagent, Biological Stain, for microscopy | Masson, Goldner, Mallory-related systems | Most paraffin-embedded tissue sections | Cell nuclei, nuclear chromatin | |
Heidenhain Iron Hematoxylin Staining Solution | BioReagent,for microscopy,Biological Stain | Histochemical nuclear staining, trichrome-related pretreatment | Tissue sections, structural localization samples | Cell nuclei, some tissue structures | |
Thomas phosphomolybdic Acid Hematoxylin Staining Solution | BioReagent,Biological Stain,for microscopy | Trichrome/histochemical-related systems | Tissue structure observation samples | Nuclear structures, tissue details | |
Nuclear Fast Red Staining Solution | BioReagent, Biological Stain, for microscopy, 0.1% | Collagen staining control, histochemical counterstaining | Tissue sections after collagen staining | Cell nuclei | |
Nuclear Fast Red Staining Solution (0.2%) | BioReagent,Biological Stain,for microscopy,0.2% | Collagen staining control, histochemical counterstaining | Tissue sections after collagen staining | Cell nuclei | |
Hematoxylin Staining Solution (Gill No.1) | BioReagent, Biological Stain, for microscopy | Histological nuclear staining | Routine tissue sections | Cell nuclei | |
Hematoxylin Staining Solution (Gill No.2) | BioReagent, Biological Stain, for microscopy | Histological nuclear staining | Routine tissue sections | Cell nuclei | |
Hematoxylin Staining Solution (Gill No.3) | BioReagent, Biological Stain, for microscopy | Histological nuclear staining | Routine tissue sections | Cell nuclei | |
Mayer hematoxylin staining solution | BioReagent, Biological Stain, for microscopy | Routine tissue nuclear staining | Tissue sections, cell samples | Cell nuclei |
5.4 Fast Green-Related Osteochondral Matrix Contrast Staining Products
Table 8 Fast Green-Related Osteochondral Matrix Contrast Staining Products
Cat. No. | Product Name | Grade/Specification | Suitable samples | Main targets |
Safranine-Fast Green Staining Kit | BioReagent, Biological Stain, for microscopy | Cartilage, osteochondral interface, joint tissue | Cartilage matrix, bone matrix, background tissue | |
Modified Safranine O-Fast Green Cartilage Staining Solution | BioReagent, Biological Stain, for microscopy, sterile | Cartilage tissue, articular cartilage, cartilage repair models | Cartilage proteoglycans, non-cartilage background matrix | |
Safranin-Fast Green Staining Solution for Bone Tissue | BioReagent,Biological Stain,for microscopy | Bone tissue, osteochondral junction, bone repair samples | Bone matrix, cartilage matrix, tissue background | |
Safranin O-Fast Green Staining Solution (for Plant Tissues) | BioReagent,for microscopy,Biological Stain | Plant tissue sections | Plant cell walls, lignified tissue, background structures | |
Cartilage Staining Solution (Safranine O) | BioReagent, Biological Stain, for microscopy | Cartilage tissue, articular cartilage, cartilage degeneration models | Cartilage proteoglycans, cartilage matrix | |
Cartilage Staining Solution (Toluidine Blue Method) | BioReagent, for microscopy, Biological Stain | Cartilage tissue, articular cartilage, osteochondral samples | Acidic mucopolysaccharides, cartilage matrix | |
Myelin Staining Solution (Fast Green Method) | BioReagent, Biological Stain, for microscopy | Neural tissue, myelin-related samples | Myelin structures, neural tissue background |
The selection of common trichrome staining systems should be based on sample type, target structure, and result purpose. Masson trichrome is suitable for collagen and fibrosis evaluation; Goldner trichrome is suitable for bone matrix and mineralized tissue observation; Mallory and Azan emphasize multistructural tissue stratification; Gomori is more oriented toward muscle pathology and special structure display; and Van Gieson can serve as a collagen control staining method.
For more related articles, please see below:
[1] Masson’s Trichrome Stain (Masson’s Trichrome) Standard Operating Procedure (SOP)