Technical articles

Principles, Classification, and Clinical Laboratory Applications of Hematologic Cytochemical Staining

Hematologic cytochemical staining is a group of detection methods that, on the basis of cellular morphology, use specific chemical reactions to demonstrate intracellular enzymes, glycogen, lipids, iron granules, and other functional components. Its value is not to replace flow cytometric immunophenotyping, cytogenetics, or molecular testing, but to provide clues about cell lineage, differentiation stage, and functional status in the preliminary differentiation of bone marrow smears, peripheral blood smears, and hematologic diseases.

 

Keywords: hematologic cytochemical staining; bone marrow smear; peripheral blood smear; peroxidase staining; Sudan Black B staining; nonspecific esterase staining; specific esterase staining; PAS staining; neutrophil alkaline phosphatase; iron staining; leukemia differentiation

 

1 Basic Positioning of Hematologic Cytochemical Staining

1.1 Detection Targets

(1) Intracellular enzyme activity

Hematologic cytochemical staining can demonstrate the activity of certain enzymes in myeloid cells, monocytes, lymphocytes, megakaryocytes, and erythroid cells, such as myeloperoxidase, specific esterase, nonspecific esterase, alkaline phosphatase, and acid phosphatase. These enzyme activities are associated with cell lineage and differentiation stage.

(2) Intracellular chemical components

Some staining methods are used to demonstrate glycogen, lipids, iron granules, and reticular-fiber-related components. For example, PAS staining is used to observe glycogen and glycoproteins, while iron staining is used to evaluate bone marrow iron stores and ring sideroblasts.

(3) Cellular functional status

Certain cytochemical reactions reflect cellular function or maturation status. For example, the neutrophil alkaline phosphatase score can help distinguish chronic myeloid leukemia from leukemoid reaction, while nonspecific esterase can suggest monocytic differentiation.

 

1.2 Methodological Characteristics

(1) Dependence on morphological background

Cytochemical staining must be interpreted together with smear morphology. The cell type showing positive granules, the staining intensity, and the percentage of positive cells all need to be evaluated microscopically in combination with cell size, nuclear morphology, cytoplasmic volume, granules, and maturation stage.

(2) Mostly semi-quantitative results

Most hematologic cytochemical staining results are reported as negative, weakly positive, positive, strongly positive, or as a score. They are essentially semi-quantitative or categorical interpretations and should not be directly equated with precise enzymatic activity quantification.

(3) Strong influence of sample quality

Bone marrow smear thickness, cellular integrity, anticoagulants, fixation conditions, staining solution freshness, incubation temperature, and reaction time can all affect staining results. Cytochemical staining requires highly consistent sample pretreatment.

Table 1 Main Types of Hematologic Cytochemical Staining

 

Staining Type

Main Detection Target

Typical Positive Cells

Main Use

Peroxidase staining (MPO/POX)

Myeloperoxidase

Granulocytic cells, some myeloid blasts

Preliminary differentiation of myeloid and lymphoid lineages

Sudan Black B staining (SBB)

Lipids, phospholipids, and myeloid granule-related components

Granulocytic cells, some monocytic cells

Auxiliary assessment of myeloid differentiation

Specific esterase staining

Chloroacetate esterase and related enzymes

Granulocytic cells

Identification of granulocytic differentiation

Nonspecific esterase staining

α-Naphthyl esterases

Monocytes, macrophages

Identification of monocytic differentiation

PAS staining

Glycogen, glycoproteins, polysaccharides

Abnormal erythroid cells, lymphoblasts, etc.

Evaluation of erythroleukemia, ALL, and some abnormal proliferations

Neutrophil alkaline phosphatase staining

Neutrophil alkaline phosphatase

Mature neutrophils

Auxiliary differentiation between CML and leukemoid reaction

Iron staining

Intracellular and extracellular iron

Bone marrow macrophages, sideroblasts

Evaluation of bone marrow iron stores and sideroblasts

Acid phosphatase staining

Acid phosphatase

Some lymphocytes, monocytes, hairy cells, etc.

Auxiliary assessment of specific lymphoproliferative diseases

 

2 Peroxidase Staining

2.1 Staining Principle

Peroxidase staining is mainly used to demonstrate myeloperoxidase activity. Myeloperoxidase is present in the azurophilic granules of granulocytic cells and some myeloid blasts. It can catalyze the oxidation of chromogenic substrates by hydrogen peroxide, forming yellow-brown, brown-black, or dark granular deposits.

The core significance of this reaction is to determine whether blasts show myeloid differentiation. In general, granulocytic cells are MPO-positive, monocytic cells may be weakly positive or negative, and lymphoid blasts are usually negative.

 

2.2 Application Scenarios

(1) Preliminary classification of acute leukemia

In the morphological diagnosis of acute leukemia, MPO positivity supports acute myeloid leukemia. MPO negativity suggests the need to consider acute lymphoblastic leukemia, undifferentiated acute leukemia, or some poorly differentiated AML. Results still need to be interpreted together with flow cytometric immunophenotyping and genetic testing.

(2) Assessment of myeloid differentiation

When some blasts show atypical morphology, MPO staining helps confirm whether myeloid granule enzyme activity is present. Positive granules are distributed in the cytoplasm and are often associated with the degree of cellular differentiation.

(3) Auxiliary evaluation of bone marrow after treatment

In post-treatment bone marrow, MPO staining can help identify regenerating granulocytic cells and residual abnormal cells, but it is not suitable as an independent method for minimal residual disease detection.

 

2.3 Interpretation Points

MPO interpretation should focus on the percentage of positive cells, granule intensity, and morphology of positive cells. Mature neutrophils are usually strongly positive. Weak positivity in blasts should be interpreted cautiously to avoid misinterpreting nonspecific deposits or background granules as true positivity.

Table 2 Interpretation of Peroxidase Staining Results

 

Cell Type

Common Reaction

Interpretation Significance

Notes

Neutrophils

Strongly positive

Feature of mature granulocytic cells

Can serve as an internal control

Myeloblasts

Negative to positive

Positivity supports myeloid differentiation

Poorly differentiated AML may be weakly positive or negative

Monocytes

Negative to weakly positive

Unstable; should be combined with NSE

Not suitable alone for determining monocytic differentiation

Lymphoblasts

Usually negative

Supports lymphoid direction

Requires immunophenotyping

Eosinophils

May be positive

Granules are prominent

Should be identified with morphology

 

3 Sudan Black B Staining

3.1 Staining Principle

Sudan Black B is a fat-soluble dye that stains intracellular lipids, phospholipids, and myeloid granule-related components. In hematologic cytochemistry, Sudan Black B is commonly used to demonstrate cytoplasmic granules in granulocytic cells, usually appearing as black or blue-black granules.

SBB has clinical uses similar to MPO, and both can be used to assess myeloid differentiation. SBB positivity usually supports granulocytic or some monocytic differentiation, while lymphoid cells are mostly negative.

 

3.2 Application Scenarios

(1) Auxiliary differentiation of acute myeloid leukemia

SBB positivity supports myeloid origin, especially when MPO results are weak or require supplementary assessment.

(2) Observation of granulocytic maturation stage

As granulocytic cells mature, SBB-positive granules usually become more prominent. Mature granulocytes are strongly positive, while early blasts may show weaker positivity.

(3) Complementary use with MPO

SBB detects lipids and granule-related components rather than MPO enzyme activity directly, so SBB and MPO results are not completely equivalent. In some cases, they confirm each other; in others, staining intensity may differ.

 

3.3 Interpretation Boundaries

SBB may show dark background staining, and lipids or nonspecific hydrophobic structures may interfere with interpretation. Results should be interpreted based on cell morphology, granule location, and control cell reactions rather than the overall color of the smear alone.

Table 3 Comparison Between MPO and SBB Staining

 

Comparison Dimension

MPO Staining

SBB Staining

Detection target

Myeloperoxidase activity

Lipids, phospholipids, and myeloid granule-related components

Typical positive cells

Granulocytic cells

Granulocytic cells, some monocytic cells

Value in myeloid assessment

High

High; often complementary to MPO

Positive appearance

Brown to dark cytoplasmic granules

Black or blue-black cytoplasmic granules

Main limitation

Enzyme activity is affected by fixation and reaction conditions

Background and nonspecific staining need to be controlled

 

4 Esterase Staining

4.1 Specific Esterase Staining

(1) Staining principle

Specific esterase staining is commonly represented by naphthol AS-D chloroacetate esterase staining and mainly demonstrates esterase activity in granulocytic cells. Positive reactions usually appear as red, purplish-red, or brown-red granular deposits localized in the cytoplasm.

(2) Application value

Specific esterase positivity supports granulocytic differentiation and can be used to identify granulocytic components in acute myeloid leukemia. Together with MPO and SBB, it forms an important cytochemical combination for assessing granulocytic differentiation.

(3) Interpretation notes

Mature granulocytes are often strongly positive, while myeloblasts may be weakly positive. If smears are overfixed or reaction conditions are inappropriate, enzyme activity may be reduced, causing false-negative results.

 

4.2 Nonspecific Esterase Staining

(1) Staining principle

Nonspecific esterase staining commonly uses α-naphthyl acetate or α-naphthyl butyrate as substrates and mainly demonstrates esterase activity associated with monocytes and macrophages. Positive reactions are often diffuse or granular cytoplasmic staining.

(2) Identification of monocytic lineage

Monocytes, immature monocytes, and some monocytoid leukemic cells often show NSE positivity. This staining is valuable in differentiating acute monocytic leukemia and acute myelomonocytic leukemia.

(3) Sodium fluoride inhibition test

Nonspecific esterase activity in monocytic cells is usually inhibited by sodium fluoride. If the NSE-positive reaction is markedly inhibited by NaF, monocytic differentiation is supported. If it is not inhibited, other cell origins or nonspecific reactions should be considered.

 

4.3 Combined Interpretation

In acute leukemia, combined use of specific esterase and nonspecific esterase helps distinguish granulocytic, monocytic, and mixed granulocytic-monocytic differentiation. Granulocytic cells are usually specific esterase-positive; monocytic cells are usually NSE-positive and NaF-inhibitable; acute myelomonocytic leukemia may show both positive cell populations.

Table 4 Application Differences of Esterase Staining

 

Staining Type

Common Substrate Direction

Main Positive Cells

Inhibition Feature

Main Use

Specific esterase

Naphthol chloroacetate esters

Granulocytic cells

NaF inhibition is usually not the core interpretation criterion

Identification of granulocytic differentiation

Nonspecific esterase

α-Naphthyl acetate, α-naphthyl butyrate

Monocytes, macrophages

Monocytic positivity is often inhibited by NaF

Identification of monocytic differentiation

Dual esterase staining

Combination of two substrate types

Granulocytic and monocytic cells

Can distinguish two types of positive reactions

Auxiliary assessment of acute myelomonocytic leukemia

 

5 PAS Staining

5.1 Staining Principle

PAS staining refers to periodic acid-Schiff staining. Periodic acid oxidizes vicinal diol structures in carbohydrates to generate aldehyde groups, which then react with Schiff reagent to form a purplish-red or magenta signal. In hematologic cells, PAS mainly demonstrates glycogen, glycoproteins, and some polysaccharide-related components.

 

5.2 Application Scenarios

(1) Acute lymphoblastic leukemia

Lymphoblasts may show PAS positivity, often with coarse granular or block-like positivity. This feature can serve as an auxiliary morphological indicator for ALL but cannot replace immunophenotyping.

(2) Erythroleukemia and abnormal erythroid proliferation

Abnormal erythroid cells may show PAS positivity, especially in cases of abnormal erythroblast proliferation. Normal erythroblasts are usually PAS-negative or weakly reactive, while abnormal positivity suggests dysplastic erythropoiesis.

(3) Megakaryocyte and platelet-related observation

Megakaryocytes and platelets may be PAS-positive, which can help identify the platelet system and some megakaryocytic abnormalities.

 

5.3 Interpretation Points

The PAS positivity pattern is more meaningful than positivity intensity alone. Diffuse, fine granular, coarse granular, or block-like positivity suggests different intracellular carbohydrate distribution patterns. Interpretation should be combined with cell lineage, morphological abnormalities, and other cytochemical results.

Table 5 Common PAS Staining Result Patterns

 

Cell Type

Common PAS Reaction

Interpretation Significance

Notes

Lymphoblasts

Granular or block-like positivity may be seen

Supports auxiliary assessment of ALL

Requires immunophenotyping

Normal erythroblasts

Mostly negative or weakly positive

Normal erythroid feature

Strong positivity suggests abnormal erythroid lineage

Abnormal erythroid cells

May be clearly positive

Supports dysplastic erythropoiesis

Should be combined with bone marrow morphology and genetics

Megakaryocytes

Often positive

Reflects abundant glycoproteins

Can help identify megakaryocytes

Granulocytic cells

May be weakly positive or nonspecific

Limited diagnostic specificity

Not a core marker for granulocytic identification

 

6 Neutrophil Alkaline Phosphatase Staining

6.1 Staining Principle

Neutrophil alkaline phosphatase staining is used to demonstrate alkaline phosphatase activity in mature neutrophils. Reaction products are usually localized in the cytoplasm of mature neutrophils, and results are generally scored based on staining intensity and the proportion of positive cells.

 

6.2 Clinical Applications

(1) Differentiation between CML and leukemoid reaction

The NAP score is usually decreased in chronic myeloid leukemia and often increased in leukemoid reaction. This indicator was once an important laboratory basis for differentiating the two.

(2) Evaluation of infection and inflammatory status

Severe infection, inflammatory stimulation, and stress states may increase neutrophil alkaline phosphatase activity, suggesting neutrophil functional activation.

(3) Auxiliary assessment of myeloproliferative diseases

NAP scoring can provide reference value for differentiating some myeloproliferative diseases, but modern diagnosis relies more on genetics, molecular markers, and comprehensive clinical assessment.

 

6.3 Interpretation Limitations

The NAP score is affected by infection, drugs, pregnancy, hormones, sample storage, and operating conditions. It cannot be used alone to diagnose CML or other myeloproliferative diseases and should be interpreted together with BCR-ABL1 testing, blood counts, bone marrow morphology, and clinical presentation.

Table 6 Common Interpretation of NAP Staining Results

 

Result Pattern

Common Background

Interpretation Direction

Notes

Decreased NAP score

Common in CML

Supports chronic myeloid leukemia direction

Requires BCR-ABL1 confirmation

Increased NAP score

Leukemoid reaction, infection, inflammation

Supports reactive granulocytosis

Should be combined with infection markers

Normal NAP score

Seen in multiple conditions

Limited diagnostic specificity

Should not be used alone to exclude disease

Strong positivity in mature neutrophils

Reactive activation

Functional activation

Strongly affected by clinical status

 

7 Iron Staining

7.1 Staining Principle

Bone marrow iron staining commonly uses the Prussian blue reaction to demonstrate ferric iron. Iron reacts with acidic ferrocyanide to form blue deposits, allowing observation of extracellular storage iron in bone marrow and intracellular iron granules in erythroblasts.

 

7.2 Application Scenarios

(1) Evaluation of bone marrow storage iron

Blue granules in bone marrow macrophages reflect storage iron status. Iron deficiency anemia often shows reduced or absent bone marrow storage iron.

(2) Observation of sideroblasts

Erythroblasts containing blue iron granules in the cytoplasm are called sideroblasts. If iron granules are arranged around the nucleus in a ring, they are called ring sideroblasts, which are important morphological clues in some myelodysplastic syndromes and sideroblastic anemia.

(3) Differentiation of anemia

Iron staining helps distinguish iron deficiency anemia, anemia of chronic disease, sideroblastic anemia, and some MDS-related anemias.

 

7.3 Interpretation Points

Iron staining requires separate evaluation of extracellular and intracellular iron. Extracellular iron reflects storage iron, while intracellular iron reflects erythroid iron utilization. Iron contamination of smears, improper decolorization, or insufficient marrow particles can all affect interpretation.

Table 7 Interpretation Directions of Bone Marrow Iron Staining

 

Observation Target

Positive Appearance

Main Significance

Common Application

Extracellular iron

Blue granules in macrophages or marrow particles

Storage iron level

Differentiation of iron deficiency anemia and anemia of chronic disease

Sideroblasts

Blue iron granules in erythroblast cytoplasm

Erythroid iron utilization status

Anemia classification and erythroid maturation evaluation

Ring sideroblasts

Iron granules arranged around the nucleus

Abnormal mitochondrial iron deposition

MDS, sideroblastic anemia

Negative or very little iron

Lack of storage iron

Supports iron deficiency

Should be combined with serum ferritin and clinical indicators

 

8 Acid Phosphatase and Other Special Stains

8.1 Acid Phosphatase Staining

Acid phosphatase staining can demonstrate acid phosphatase activity in some lymphocytes, monocytes, macrophages, and certain abnormal cells. It has auxiliary value in diseases such as hairy cell leukemia. Tartrate-resistant acid phosphatase (TRAP) reaction was once used as an auxiliary diagnostic method for hairy cell leukemia.

 

8.2 β-Glucuronidase Staining

β-Glucuronidase staining can reflect enzyme activity associated with parts of the monocyte-macrophage system and can be used as a supplementary indicator for monocytic differentiation, although its routine clinical use is less frequent than MPO, SBB, esterase, and PAS staining.

 

8.3 Enzyme Stains Other Than Alkaline Phosphatase

Some laboratories may also perform acid esterase, phosphatase, dehydrogenase, and other stains for specific research or auxiliary disease assessment. These methods require clearly defined detection purposes, positive controls, and interpretation criteria.

 

9 Combined Application of Hematologic Cytochemical Staining

9.1 Preliminary Differentiation of Acute Leukemia

In acute leukemia, MPO, SBB, specific esterase, nonspecific esterase, and PAS are commonly used to preliminarily distinguish myeloid, monocytic, and lymphoid differentiation. MPO or SBB positivity supports myeloid differentiation; NSE positivity with NaF inhibition supports monocytic differentiation; coarse granular or block-like PAS positivity may suggest lymphoblasts or abnormal erythroid features.

 

9.2 Assessment of AML Subtypes

In acute granulocytic leukemia, MPO, SBB, and specific esterase are often positive. In acute monocytic leukemia, nonspecific esterase positivity is more meaningful. In acute myelomonocytic leukemia, cytochemical features of both granulocytic and monocytic lineages may coexist.

 

9.3 Evaluation of Anemia and Erythroid Abnormalities

Iron staining and PAS staining can be used for anemia classification and evaluation of abnormal erythroid proliferation. Iron staining focuses on storage iron, sideroblasts, and ring sideroblasts; PAS staining may suggest dysplastic erythropoiesis and some erythroleukemia-related changes.

 

9.4 Auxiliary Assessment of Myeloproliferative Diseases

NAP staining can serve as an auxiliary method for evaluating neutrophil functional status and reactive granulocytosis. Chronic myeloid leukemia often shows a reduced NAP score, but modern diagnosis must be combined with molecular tests such as BCR-ABL1.

Table 8 Cytochemical Staining Combinations in Common Disease Scenarios

 

Disease or Scenario

Recommended Staining Combination

Typical Clues

Interpretation Focus

Acute myeloid leukemia

MPO, SBB, specific esterase

Myeloid cells positive

Requires flow cytometric immunophenotyping

Acute monocytic leukemia

NSE, NaF inhibition test, MPO

NSE-positive and inhibited by NaF

Distinguish monocytic from granulocytic differentiation

Acute myelomonocytic leukemia

Specific esterase, NSE, MPO

Granulocytic and monocytic reactions coexist

Focus on different positive cell populations

Acute lymphoblastic leukemia

PAS, MPO, SBB

PAS may be positive; MPO/SBB mostly negative

Cannot replace immunophenotyping

Iron deficiency anemia

Iron staining

Reduced or absent storage iron

Combine with ferritin and red cell indices

MDS with ring sideroblasts

Iron staining, PAS

Increased ring sideroblasts

Combine with morphology and genetics

CML vs leukemoid reaction

NAP staining

Low in CML, high in leukemoid reaction

Requires BCR-ABL1 confirmation

 

10 Quality Control and Common Problems

10.1 Smear Quality

Hematologic cytochemical staining requires smears of appropriate thickness, uniform cell distribution, and intact cell morphology. Excessively thick smears may cause uneven staining and high background; overly thin smears may contain too few positive cells, making scoring and classification difficult.

 

10.2 Fixation Conditions

Different cytochemical stains have different sensitivities to fixatives. Enzyme stains in particular require avoidance of overfixation, which can reduce enzyme activity. Fixation time, fixative type, and sample drying status should be strictly controlled according to the specific staining method.

 

10.3 Control Setup

Positive and negative controls should be included in each staining run. Mature neutrophils can serve as internal controls for MPO and SBB. Known positive samples can be used for quality control of esterase, PAS, NAP, and iron staining.

 

10.4 Result Interpretation

Cytochemical staining results should be interpreted together with cellular morphology, peripheral blood findings, bone marrow morphology, flow cytometric immunophenotyping, karyotyping, and molecular testing. A single positive or negative stain cannot independently establish a diagnosis of hematologic malignancy.

Table 9 Common Problems and Optimization Directions in Hematologic Cytochemical Staining

 

Problem

Possible Cause

Impact on Results

Optimization Direction

Overall weak enzyme staining

Overfixation, reagent failure, insufficient reaction time

Risk of false negatives

Optimize fixation and include positive controls

Deep background staining

Smear too thick, insufficient washing, stain precipitation

Difficult interpretation of positive granules

Improve smear quality and filter staining solution

Unclear positive granules

Improper reaction time, unstable chromogenic substrate

Difficult grading

Control incubation time and temperature

Large fluctuation in NAP score

Improper sample storage, insufficient cells counted

Poor comparability between groups

Standardize sampling and counting rules

False-positive iron staining

Exogenous iron contamination, slide contamination

Overestimation of storage iron

Use clean instruments and include blanks

Inconsistent PAS results

Unstable oxidation time or Schiff reagent condition

Bias in positivity intensity

Control periodic acid oxidation time and reagent freshness

 

11 Reagent Selection Related to Hematologic Cytochemical Staining

Table 10 Common Basic Reagents and Materials for Hematologic Cytochemical Staining

 

Product Category

Product Name

CAS No.

Role in the System

Applicable Direction

Peroxidase chromogenic substrate

3,3'-Diaminobenzidine (DAB)

91-95-2

Peroxidase chromogenic substrate

MPO/POX staining, enzyme histochemical visualization

Lipid dye

Sudan Black B

4197-25-5

Stains lipids, phospholipids, and myeloid granule-related components

SBB staining, auxiliary assessment of myeloid differentiation

Lipid dye

Sudan III

85-86-9

Demonstrates neutral lipids

Observation of lipid granules and fat components

Lipid dye

Sudan IV

85-83-6

Demonstrates neutral lipids

Lipid deposition and lipid droplet observation

PAS oxidant

Periodic acid

10450-60-9

Oxidizes vicinal diols in carbohydrates to form aldehyde groups

Oxidation step before PAS staining

Glycogen digestion control

Amylase

9000-90-2

Digests glycogen to verify the source of PAS positivity

PAS staining quality control and specificity validation

Iron staining reagent

Potassium ferrocyanide

14459-95-1

Forms Prussian blue reaction with ferric iron

Bone marrow iron staining

Nonspecific esterase substrate

α-Naphthyl acetate

830-81-9

NSE reaction substrate

Monocyte- and macrophage-related esterase staining

Nonspecific esterase substrate

α-Naphthyl butyrate

3121-70-8

NSE reaction substrate

Auxiliary assessment of monocytic differentiation

Specific esterase substrate

Naphthol AS-D chloroacetate

35245-26-2

Specific esterase reaction substrate

Identification of granulocytic differentiation

Azo coupling salt

Fast Blue RR Salt

14726-29-5

Couples with naphthol products for color development

Esterase and phosphatase staining

Azo coupling salt

Fast Garnet GBC Salt

101-89-3

Azo coupling chromogen

Esterase and phosphatase histochemistry

Azo coupling salt

Fast Red Violet LB Salt

32348-81-5

Forms red deposits by coupling

Esterase and phosphatase staining

Esterase inhibitor

Sodium fluoride

7681-49-4

Inhibits monocytic nonspecific esterase

NSE sodium fluoride inhibition test

Alkaline phosphatase substrate

Naphthol AS-BI phosphate

1919-91-1

Alkaline phosphatase chromogenic substrate

NAP staining

TRAP inhibition/differentiation reagent

L-Tartaric acid

87-69-4

Used for tartrate-resistant acid phosphatase differentiation

TRAP staining, auxiliary assessment of hairy cell leukemia

 

Table 11 Ready-to-Use Reagent Kits and Auxiliary Materials for Hematologic Cytochemical Staining

 

Product Category

Cat. No.

Product Name

Grade / Specification

Role in the System

Applicable Direction

PAS chromogenic reagent

S1516056

Schiff Reagent

BioReagent,Biological Stain,for microscopy

Reacts with aldehyde groups generated after periodic acid oxidation to produce a magenta signal

PAS staining, glycogen and glycoprotein demonstration; suitable for hematologic cytochemical PAS systems

Peroxidase staining

P1518603

Peroxidase Staining Solution (Oxidase WG-KI Method)

BioReagent,Biological Stain,for microscopy

Demonstrates intracellular peroxidase-related reactions

MPO/POX staining, auxiliary assessment of myeloid differentiation in acute leukemia

Peroxidase staining

P1518605

Peroxidase Staining Solution (Benzidine Method)

BioReagent,Biological Stain,for microscopy

Peroxidase catalyzes substrate oxidation to form chromogenic deposits

Granulocytic cell identification and auxiliary differentiation of myeloid blasts

Sudan Black B staining

S1508362

Sudan Black B Staining Solution

BioReagent,for microscopy,Biological Stain

Demonstrates intracellular lipids, phospholipids, and myeloid granule-related components

SBB staining, auxiliary differentiation between AML and ALL

Sudan Black B staining

S1508441

Sudan Black B Staining Solution

BioReagent,for microscopy,Biological Stain

Forms black or blue-black granular staining signals

Granulocytic differentiation, myeloid granule observation, hematologic cytochemical staining

PAS combined staining

A774824

AB-PAS staining kit

BioReagent, for microscopy, Biological Stain

Simultaneously demonstrates acidic mucosubstances and PAS-positive components

Observation of carbohydrate components in tissues/cells; extended PAS-related system

Glycogen PAS staining

G774821

Glycogen D-PAS Staining Solution (Amylase Digestion)

BioReagent, for microscopy, Biological Stain

Uses amylase digestion to verify whether PAS positivity originates from glycogen

Quality control for hematologic PAS staining and specificity validation of glycogen positivity

Glycogen PAS staining

G774820

Glycogen PAS staining kit

BioReagent, Biological Stain, for microscopy

Demonstrates glycogen, glycoproteins, and polysaccharide components

Observation of PAS positivity patterns in ALL, abnormal erythroid proliferation, erythroleukemia, etc.

Glycogen PAS staining

G774823

Glycogen PAS Staining Kit (Special For Cell)

BioReagent, Biological Stain, for microscopy

Optimized PAS staining system for cultured cell samples

Observation of glycogen and glycoprotein components in cultured hematologic cells or cell lines

Prussian blue iron staining

P113746

Prussian blue

Biological Stain

Chromogenic material related to iron staining

Iron staining-related material; suitable for method expansion in iron deposition observation

Prussian blue iron staining

P131065

Prussian blue soluble

Biological Stain

Chromogenic material related to iron staining

Prussian blue-related staining systems and iron deposition observation

Prussian blue iron staining

P1508698

Prussian Blue Staining Solution (DAB Enhancement Method)

BioReagent,for microscopy,Biological Stain

Enhances Prussian blue iron staining signals

Bone marrow iron staining, observation of weakly positive iron granules, enhanced iron deposition visualization

Prussian blue iron staining

P774842

Prussian Blue Staining Kit (Neutral Red)

BioReagent, Biological Stain, for microscopy

Demonstrates ferric iron and uses neutral red counterstaining for localization

Observation of bone marrow storage iron, sideroblasts, and tissue iron deposition

Prussian blue iron staining

P774832

Prussian Blue Staining Kit (Eosin)

BioReagent, Biological Stain, for microscopy

Combines Prussian blue iron reaction with eosin counterstaining

Bone marrow or tissue iron staining; auxiliary cellular morphology localization

Prussian blue iron staining

P774831

Prussian Blue Staining Kit (Nuclear Fast Red)

BioReagent, for microscopy, Biological Stain

Combines Prussian blue iron reaction with nuclear fast red counterstaining

Bone marrow iron staining, sideroblast identification, tissue iron deposition localization

Basic morphology staining

M743374

Wright-Giemsa Staining Kit

BioReagent, Biological Stain, for microscopy

Provides basic morphological background for blood and bone marrow cells

Combined interpretation with MPO, SBB, PAS, iron staining, and other cytochemical results

Mounting material

F1508973

Fischor Mounting Medium

BioReagent,for microscopy,Suitable for Immunofluorescence(IF),Suitable for Immunohistochemistry(IHC)

Mounting after staining to maintain microscopic observation condition

Hematologic cytochemical staining, histological staining, and image acquisition

Mounting material

M292692

mounting medium

Unscented

Routine mounting

Microscopic observation after smear or tissue section staining

Glycerol mounting

C1508971

Routine Glycerol Mounting Medium

BioReagent,for microscopy,Suitable for Immunofluorescence(IF),for fluorescence analysis

Aqueous or semi-aqueous mounting to preserve stained samples

Cell smears, fluorescence combined staining, or temporary observation

Glycerol-PBS mounting

G1508970

Glycerol PBS Mounting Medium

BioReagent,for microscopy,Suitable for Immunofluorescence(IF),for fluorescence analysis

Buffered glycerol mounting system

Smear observation after staining, fluorescent or enzyme-staining combined samples

Antifade mounting

A752105

Enhanced Antifade Mounting Medium

BioReagent, for fluorescence analysis, Suitable for Immunofluorescence(IF)

Reduces fluorescence signal decay

Cytochemical staining combined with immunofluorescence or fluorescent labeling experiments

Antifade mounting

A598329

Antifluorescent quencher

 

Protects fluorescence signal and reduces quenching

Fluorescent cytochemical staining and immunofluorescence combined observation

Antifade mounting

P1508975

Polyvinyl Alcohol Anti-Fluorescence Quenching Mounting Medium

Suitable for Immunofluorescence(IF),BioReagent,for microscopy,for fluorescence analysis

Improves storage stability of fluorescent samples

Fluorescent staining samples and immunofluorescence-combined cytochemical experiments

Glycerol mounting

P1508974

Polyvinyl Alcohol Glycerol Mounting Medium

BioReagent,for microscopy,Suitable for Immunofluorescence(IF),Suitable for Immunohistochemistry(IHC)

Provides a relatively stable mounting environment

Blood smears, tissue sections, and immunostained samples

Glycerol mounting

A1508972

Gum Arabic Glycerol Mounting Medium

BioReagent,for microscopy,Suitable for microbiology,Suitable for Immunofluorescence(IF),Suitable for Immunohistochemistry(IHC)

Traditional glycerol-based mounting system

Mounting and long-term observation of microscopic staining samples

 

The core value of hematologic cytochemical staining lies in combining cellular morphology with demonstration of functional components, providing directional evidence for the preliminary differentiation of hematologic diseases. In practice, smear quality, fixation conditions, positive controls, and interpretation standards should be kept consistent, and the results should be incorporated into an integrated diagnostic framework including morphology, immunophenotype, genetics, and molecular testing.

Categories: Technical articles

Da — when not otherwise indicated, molecular weight units are daltons.   Mw — weight-average molecular weight.   Mn — number-average molecular weight.

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Cite this article

Aladdin Scientific. "Principles, Classification, and Clinical Laboratory Applications of Hematologic Cytochemical Staining" Aladdin Knowledge Base, updated May 26, 2026. https://www.aladdinsci.com/us_en/faqs/principles-classification-and-clinical-laboratory-applications-of-hematologic-cytochemical-staining-en.html
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