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E-raamat: Basic and Advanced Laboratory Techniques in Histopathology and Cytology

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  • Formaat: EPUB+DRM
  • Sari: Medicine
  • Ilmumisaeg: 08-Jun-2018
  • Kirjastus: Springer Verlag, Singapore
  • Keel: eng
  • ISBN-13: 9789811082528
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Basic and Advanced Laboratory Techniques in Histopathology and CytologySuurem pilt
  • Formaat: EPUB+DRM
  • Sari: Medicine
  • Ilmumisaeg: 08-Jun-2018
  • Kirjastus: Springer Verlag, Singapore
  • Keel: eng
  • ISBN-13: 9789811082528
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This book provides detailed information on basic and advanced laboratory techniques in histopathology and cytology. It discusses the principles of and offers clear guidance on all routine and special laboratory techniques. In addition, it covers various advanced laboratory techniques, such as immunocytochemistry, flow cytometry, liquid based cytology, polymerase chain reaction, tissue microarray, and molecular technology. Further, the book includes numerous color illustrations, tables and boxes to familiarize the reader with the work of a pathology laboratory.

 

The book is mainly intended for postgraduate students and fellows in pathology as well as practicing pathologists. The book is also relevant for all the laboratory technicians and students of laboratory technology.

Part I Basic Laboratory Techniques in Histopathology Laboratory
1 Fixation of Histology Samples: Principles, Methods and Types of Fixatives
3(16)
1.1 Introduction
3(1)
1.2 Aims of Fixation
3(1)
1.3 Ideal Fixative
3(1)
1.4 Tissue Changes in Fixation
3(1)
1.5 Types of Fixation
4(2)
1.5.1 Description of Nature of Fixation
5(1)
1.6 Essential Precautions for Fixation in General
6(1)
1.7 Mechanism of Fixation
6(2)
1.8 Factors Affecting Fixation
8(2)
1.9 Commonly Used Fixatives in the Laboratory
10(3)
1.9.1 Formaldehyde
10(1)
1.9.2 Preparation of Different Formalin Solution
10(1)
1.9.3 Glutaraldehyde
11(1)
1.9.4 Osmium Tetroxide
12(1)
1.9.5 Methyl and Ethyl Alcohol
12(1)
1.9.6 Acetone
12(1)
1.9.7 Bouin's Fixative
12(1)
1.10 Mercury Salt-Containing Fixatives
13(2)
1.10.1 Zenker's Fluid
13(1)
1.10.2 Helly's Fluid
13(1)
1.10.3 B5 Fixatives
13(1)
1.10.4 Fixatives of Choice
13(2)
1.11 Fixation Artefact
15(2)
References
17(2)
2 Processing of Tissue in Histopathology Laboratory
19(10)
2.1 Factors that Influence Tissue Processing
19(1)
2.2 Dehydration
20(1)
2.3 Individual Dehydrating Agent
21(1)
2.3.1 Alcohol
21(1)
2.3.2 Dehydrating Agents Other than Alcohol
21(1)
2.4 Clearing
22(2)
2.4.1 Individual Clearing Agent
22(1)
2.4.2 Other Clear Agents
23(1)
2.5 Infiltration and Embedding
24(1)
2.5.1 Different Impregnating Medium
24(1)
2.6 Tissue Processing Methods
25(1)
2.7 Overall Precautions of Tissue Processing
26(1)
2.7.1 Time Schedule for Overnight Processing
26(1)
2.7.2 Manual Tissue Processor
26(1)
2.7.3 Microwave Processing
27(1)
References
27(2)
3 Embedding of Tissue in Histopathology
29(6)
3.1 Embedding Medium
29(1)
3.2 Different Types of Mould Used for Block
30(1)
3.3 Tissue Embedding Method
30(2)
3.4 Tissue Orientation and Embedding
32(1)
3.5 Tissue Marking
32(1)
References
33(2)
4 Decalcification of Bony and Hard Tissue for Histopathology Processing
35(6)
4.1 Introduction
35(1)
4.1.1 Factors Controlling the Rate of Decalcification
36(1)
4.2 The Methods of Decalcification
36(1)
4.3 Chelating Agents
37(1)
4.3.1 Other Procedures of Decalcification
38(1)
4.4 Surface Decalcification
38(1)
4.5 End Point Determination of Decalcification
39(1)
Reference
39(2)
5 Tissue Microtomy: Principle and Procedure
41(10)
5.1 Introduction
41(1)
5.2 Microtomes
41(3)
5.2.1 Microtome Knife
43(1)
5.2.2 Disposable Knife
44(1)
5.2.3 Materials Used in Knife
44(1)
5.2.4 Angles of Knife
44(1)
5.3 Microtome Knife Sharpening
44(1)
5.3.1 Manual Method
44(1)
5.3.2 Factors Involved in Cutting
45(1)
5.4 Sectioning the Paraffin Block
45(5)
5.4.1 Steps of Tissue Sectioning
46(4)
Reference
50(1)
6 Frozen Section: Principle and Procedure
51(6)
6.1 Introduction
51(1)
6.2 Indications of Frozen Sections
51(1)
6.3 The Principle of Frozen Section
51(1)
6.4 Cryostat Sectioning
52(2)
6.5 Staining
54(1)
6.5.1 H&E Staining
54(1)
6.5.2 Toluidine Blue Stain
55(1)
6.6 Factors Affecting the Good-Quality Section
55(1)
References
55(2)
7 Staining Principle and General Procedure of Staining of the Tissue
57(12)
7.1 Introduction
57(2)
7.1.1 Dyes Used for Staining
57(1)
7.1.2 Types of Dye
58(1)
7.1.3 Types of Dye Based on Chemical Structures and Chromophore Groups
59(1)
7.2 Mechanisms and Theory of Staining
59(3)
7.3 Factors Influencing Staining
62(1)
7.3.1 Nomenclature Used Regarding Dye
62(1)
7.4 Metachromasia
63(2)
7.4.1 Metachromatic Dyes
63(2)
7.5 Progressive and Regressive Staining
65(1)
7.6 Mordant
65(1)
7.6.1 Accentuators
66(1)
7.7 Staining Procedure
66(1)
7.7.1 Preparation of Buffer Solutions
67(1)
References
67(2)
8 Haematoxylin and Eosin Stain of the Tissue Section
69(12)
8.1 Introduction
69(1)
8.2 Haematoxylin
69(1)
8.3 Bluing
70(2)
8.3.1 Preparation of Different Haematoxylins and Their Properties
71(1)
8.3.2 Mayer's Haematoxylin
71(1)
8.3.3 Ehrlich's Haematoxylin
71(1)
8.3.4 Cole's Haematoxylin
72(1)
8.4 Counterstain by Eosin
72(1)
8.5 Routine Haematoxylin and Eosin Stain
72(1)
8.6 Iron Haematoxylin
73(4)
8.6.1 Heidenhain's Iron Haematoxylin
75(1)
8.6.2 Verhoeff's Iron Haematoxylin
76(1)
8.6.3 Tungsten Haematoxylin
76(1)
8.7 Clearing of the Smear
77(2)
8.7.1 Mounting
77(2)
8.7.2 Coverslip
79(1)
References
79(2)
9 Special Stains for the Carbohydrate, Protein, Lipid, Nucleic Acid and Pigments
81(18)
9.1 Introduction
81(1)
9.2 Carbohydrates
81(3)
9.2.1 Simple Carbohydrates
82(2)
9.3 Staining of Different Carbohydrates
84(4)
9.3.1 Glycogen
84(3)
9.3.2 Combined PAS-Alcian Blue Staining
87(1)
9.4 Lipids
88(1)
9.5 Stains
89(2)
9.5.1 Oil RedO
89(1)
9.5.2 Sudan Black B
90(1)
9.5.3 Ferric Haematoxylin for Phospholipid
91(1)
9.6 Nucleic Acid and Proteins
91(1)
9.6.1 Nucleic Acids
91(1)
9.6.2 Proteins
91(1)
9.6.3 Feulgen Stain
91(1)
9.6.4 Methyl Green-Pyronin Stain
92(1)
9.7 Pigments
92(1)
9.8 Hemosiderin Pigment
92(1)
9.8.1 Prussian Blue Reaction (Perls' Reaction) for Ferric Iron
92(1)
9.9 Bile Pigment
93(1)
9.9.1 Fouchet's Stain
93(1)
9.10 Argyrophil Pigments
94(3)
9.10.1 Grimelius Staining
94(1)
9.10.2 Melanin
95(1)
9.10.3 Schmorl's Stain
95(1)
9.10.4 Calcium
96(1)
9.10.5 Formalin Pigment
97(1)
References
97(2)
10 Connective Tissue Stain: Principle and Procedure
99(10)
10.1 Fibrous Part of Connective Tissue
99(1)
10.1.1 Collagen
99(1)
10.1.2 Reticulin Fibres
100(1)
10.1.3 Elastic Fibres
100(1)
10.1.4 Basement Membrane
100(1)
10.2 Stains
100(5)
10.2.1 Masson Trichrome
100(2)
10.2.2 Van Gieson Stain
102(1)
10.2.3 Reticulin Stain
102(1)
10.2.4 Gordon and Sweet's Method for Reticulin Stain
103(2)
10.3 Elastic Fibres
105(3)
10.3.1 Verhoeff's Stain for Collagen
105(1)
10.3.2 Weigert's Resorcin-Fuchsin Stain
106(1)
10.3.3 Orcein for Elastic Fibres
106(1)
10.3.4 Phosphotungstic Acid Haematoxylin (PTAH)
106(2)
References
108(1)
11 Amyloid Staining
109(4)
11.1 Introduction
109(1)
11.2 Stains for Amyloid
110(1)
11.2.1 Alkaline Congo Red Stain
110(1)
11.2.2 Congo Red Stain by Highman
110(1)
11.2.3 Thioflavine T Stain
111(1)
References
111(2)
12 Stains for the Microbial Organisms
113(8)
12.1 Bacteria
113(3)
12.1.1 Gram's Stain
113(1)
12.1.2 Ziehl-Neelsen Stain
114(1)
12.1.3 Fite Acid-Fast Stain for Leprosy
115(1)
12.2 Fungal Infection
116(1)
12.2.1 Grocott's Methenamine Silver
116(1)
12.3 Spirochaetes
117(1)
12.3.1 Warthin and Starry Technique
117(1)
12.4 Viral Inclusions
118(1)
12.4.1 Phloxine-Tartrazine Stain
118(1)
References
118(3)
Part II Basic Laboratory Techniques in Cytology Laboratory
13 Cytology Sample Procurement, Fixation and Processing
121(12)
13.1 Introduction
121(1)
13.2 Sample Collection
121(4)
13.2.1 Cervical Cytology
121(3)
13.2.2 Respiratory Samples
124(1)
13.3 Fixation
125(2)
13.3.1 Special Fixatives
126(1)
13.4 Processing of Laboratory Samples
127(1)
13.4.1 Receiving the Sample
127(1)
13.4.2 Glass Slides and Liquid Sample
127(1)
13.5 Processing
128(4)
13.5.1 Processing of Sputum
128(1)
13.5.2 Processing of Fluid: Urine, Body Fluids and Lavage
128(2)
13.5.3 Millipore Filtration
130(1)
13.5.4 Processing of Haemorrhagic Fluid
130(1)
13.5.5 Cell Block
130(1)
13.5.6 Compact Cell Block Technique
131(1)
References
132(1)
14 Routine Staining in Cytology Laboratory
133(6)
14.1 Papanicolaou's Stain
133(3)
14.1.1 Dyes Used in Papanicolaou's Staining
133(1)
14.1.2 Principle of Basic Steps
133(1)
14.1.3 Papanicolaou's Staining Steps
134(2)
14.1.4 Bluing Solution
136(1)
14.2 Precautions to Be Taken in Papanicolaou's Staining
136(1)
14.3 May Grunwald Giemsa Stain
137(1)
Reference
138(1)
15 Basic Technique of Fine Needle Aspiration Cytology
139(10)
15.1 Introduction
139(1)
15.2 Technique Proper
140(1)
15.3 Fine Needle Aspiration Procedure
141(1)
15.4 Fine Needle Sampling
142(1)
15.5 FNAC of Deep-Seated Lesions
143(2)
15.5.1 USG-Guided FNAC
144(1)
15.5.2 CT-Guided FNAC
144(1)
15.5.3 Endoscopic Ultrasound-Guided FNAC (EUS-FNAC)
144(1)
15.5.4 Complications of Guided FNAC
145(1)
15.6 Transrectal FNAC of the Prostate
145(1)
References
146(3)
Part III Advanced Techniques in Histology and Cytology Laboratories
16 Immunocytochemistry in Histology and Cytology
149(22)
16.1 Introduction
149(2)
16.1.1 Basic Principles
149(1)
16.1.2 Basic Immunology
149(2)
16.2 Detection System
151(3)
16.2.1 Peroxidase-Antiperoxidase Method
151(1)
16.2.2 Avidin and Biotin Method
152(1)
16.2.3 Avidin and Biotin Conjugated Procedure
152(1)
16.2.4 Biotin-Streptavidin Method
153(1)
16.2.5 Alkaline Phosphatase-Antialkaline Phosphatase Method
153(1)
16.2.6 Polymer-Based Labelling Method
153(1)
16.2.7 Catalysed Signal Amplification (Tyramine Signal Amplification)
154(1)
16.3 The Sample of Tissues for Immunocytochemistry
154(5)
16.3.1 Histopathology
154(1)
16.3.2 Cytology
154(1)
16.3.3 Sample Collection
154(3)
16.3.4 Fixation
157(1)
16.3.5 Antigen Retrieval
157(1)
16.3.6 Microwave Retrieval
157(1)
16.3.7 Pressure Cooker Heating
158(1)
16.3.8 Water Bath Heating
158(1)
16.4 Immunocytochemistry Technique
159(2)
16.4.1 Control
159(1)
16.4.2 Steps
159(2)
16.4.3 Chromogen
161(1)
16.5 Troubleshooting in Immunocytochemistry
161(1)
16.6 Applications
162(1)
16.7 Diagnostic Immunocytochemistry
162(1)
16.7.1 Mesothelial Cells Versus Adenocarcinoma
162(1)
16.7.2 Mesothelial Markers
162(1)
16.7.3 Adenocarcinoma Markers in Effusion Fluid
163(1)
16.8 Different Epithelial Markers
163(1)
16.9 Mesenchymal Markers
163(2)
16.10 Neuroendocrine Markers
165(1)
16.11 Lymphoid Markers
165(1)
16.12 Melanoma Markers
165(1)
16.13 Germ Cell Markers
165(1)
16.14 Site-Specific Antibody in Different Epithelial Malignancies
166(1)
16.15 Immunocytochemistry of Round Cell Tumor
166(1)
16.16 Immunocytochemistry for Therapy and Management
167(2)
16.16.1 Breast Carcinoma
167(1)
16.16.2 Gastrointestinal Stromal Tumor
168(1)
16.16.3 Lung Carcinoma
168(1)
References
169(2)
17 Flow Cytometry: Basic Principles, Procedure and Applications in Pathology
171(14)
17.1 Introduction
171(1)
17.2 Principle of Flow Cytometry
171(1)
17.3 Dye Used
171(2)
17.3.1 Fluorochrome Dyes for FCM
171(1)
17.3.2 Fluorochrome Dye for Nucleic Acid
172(1)
17.4 Samples for Flow Cytometry
173(3)
17.4.1 Cytology Samples
173(1)
17.4.2 Histology Samples
174(1)
17.4.3 Control
174(1)
17.4.4 Sample Processing
174(1)
17.4.5 Flow Cytometric Immunophenotyping (FCI)
175(1)
17.4.6 Data Acquisition
176(1)
17.5 Targets of Application
176(1)
17.6 DNA Content and Ploidy Analysis
177(1)
17.6.1 Clinical Application
178(1)
17.6.2 Diagnosis
178(1)
17.6.3 Prognosis of the Patients
178(1)
17.7 Immunophenotyping
178(4)
17.7.1 Limitations of FCI
180(1)
17.7.2 Flow Cytometry Features of Different Lymphomas
181(1)
17.7.3 Apoptosis
181(1)
17.7.4 Assessment of Sub-GI Fraction of Apoptotic Cells
181(1)
17.7.5 Apoptosis Detection by Annexin V Assay
182(1)
References
182(3)
18 Digital Image Analysis and Virtual Microscopy in Pathology
185(8)
18.1 Introduction
185(1)
18.2 Basic Principles of Image Analysis
186(1)
18.3 Details of the Image Analysis Steps
186(3)
18.4 Morphologic Features
189(1)
18.5 The Current Problems of Digital Image Analysis
190(1)
18.6 Virtual Slide and Web-Based Teaching
191(1)
18.6.1 Advantage of Virtual Slides
191(1)
18.6.2 Disadvantages of Virtual Slides
191(1)
References
192(1)
19 Liquid-Based Cytology and Automated Screening Devices in Cytology Sample
193(8)
19.1 Introduction
193(1)
19.1.1 Advantages of LBC Over Conventional Smear
193(1)
19.1.2 Limitations of Liquid-Based Cytology
194(1)
19.2 Sample Processing
194(2)
19.2.1 ThinPrep (Cytic, UK)
194(1)
19.2.2 SurePathTest
195(1)
19.3 Comparison of These Two Techniques
196(1)
19.4 Automated Screening Devices
196(2)
19.4.1 BD FocalPoint GS Imaging System
197(1)
19.4.2 Hologic ThinPrep Imaging System
197(1)
19.4.3 Comparison of Manual and Automated Devices
198(1)
References
198(3)
20 Polymerase Chain Reaction: Principle, Technique and Applications in Pathology
201(12)
20.1 Introduction
201(1)
20.1.1 What Is PCR and How It Works?
201(1)
20.2 Steps of PCR
201(1)
20.2.1 Essential Ingredients of PCR
202(1)
20.3 Procedure Proper
202(3)
20.3.1 Basic Precautions
202(1)
20.3.2 Equipment
203(1)
20.3.3 Thermal Cycling
203(1)
20.3.4 Troubleshooting
204(1)
20.3.5 Enhancing PCR Products Formation
205(1)
20.4 Types of PCR
205(4)
20.5 Applications of PCR
209(1)
References
210(3)
21 Fluorescent In Situ Hybridization Techniques in Pathology: Principle, Technique and Applications
213(8)
21.1 Introduction
213(1)
21.1.1 The Principles of FISH
214(1)
21.2 Steps to Do FISH
214(1)
21.3 Troubleshooting
215(5)
21.3.1 Different Types of FISH
215(3)
21.3.2 CGH Method
218(1)
21.3.3 Array-Based CGH
219(1)
References
220(1)
22 Tissue Microarray in Pathology: Principal, Technique and Applications
221(6)
22.1 Introduction
221(1)
22.2 Tissue Microarray Technique
221(1)
22.3 TMA Construction and Generation of Grid
221(1)
22.4 Designing the Grid
222(2)
22.5 Limitations of TMA
224(1)
22.6 Clinical Applications of TMA
225(1)
References
225(2)
23 Sanger Sequencing and Next-Generation Gene Sequencing: Basic Principles and Applications in Pathology
227(8)
23.1 Sanger Sequencing
227(1)
23.2 Next-Generation Sequencing
228(2)
23.2.1 Scope of NGS
230(1)
23.2.2 Limitations
230(1)
23.3 Comparison of Sanger Sequencing and NGS
230(1)
References
231(4)
Part IV Microscopy, Quality Control and Laboratory Organization
24 Compound Light Microscope and Other Different Microscopes
235(10)
24.1 Light
235(1)
24.2 Colours
235(1)
24.3 Image Generation and Human Vision
236(2)
24.4 Anatomical Components of a Light Microscope
238(1)
24.5 Optical Components
239(2)
24.5.1 The Major Aberrations of the Lens
240(1)
24.6 How to Take Care and Handle Your Microscope
241(1)
24.7 Other Types of Microscope
242(1)
24.7.1 Dark-Field Microscope
242(1)
24.7.2 Bright-Field Microscope
242(1)
24.7.3 Phase Contrast Microscope
242(1)
References
243(2)
25 Fluorescence and Confocal Microscope: Basic Principles and Applications in Pathology
245(8)
25.1 Transmitted Fluorescent Microscope
245(1)
25.2 Incident Fluorescent Microscope
246(1)
25.3 Confocal Microscopy
247(2)
25.4 Limitations of CFM
249(1)
25.5 Applications of CFM
249(1)
25.6 Two-Photon Microscopy
250(1)
25.7 4Pi Microscopy
250(1)
25.8 Spatially Modulated Illumination Microscopy
251(1)
References
252(1)
26 Electron Microscopy: Principle, Components, Optics and Specimen Processing
253(10)
26.1 Introduction
253(3)
26.1.1 Essential Components of Electron Microscope
254(2)
26.1.2 Microscope Column and Electronic Optics
256(1)
26.2 Specimen and Electron Interaction
256(2)
26.2.1 Electron Interaction in Transmission Electron Microscope
258(1)
26.3 Sample Preparation for TEM
258(2)
26.3.1 Sample Collection
258(1)
26.3.2 Fixation
258(1)
26.3.3 Dehydration
259(1)
26.3.4 Embedding
259(1)
26.4 Sectioning
260(2)
26.4.1 Staining of the Sections
261(1)
26.5 Scanning Electron Microscopy
262(1)
26.5.1 Operational Principle
262(1)
References
262(1)
27 Quality Control and Laboratory Organization
263(8)
27.1 Introduction
263(2)
27.2 Quality Control
265(3)
27.2.1 Pre-analytic Phase
265(1)
27.2.2 Laboratory Processing
266(1)
27.2.3 Tissue and Sample Processing
266(1)
27.2.4 Analytic Phase
266(1)
27.2.5 Post-analytic Phase
267(1)
27.2.6 Gold Standard
267(1)
27.2.7 Record-Keeping
267(1)
27.3 External Quality Assurance
268(1)
27.4 Laboratory Organization
268(1)
27.4.1 Laboratory Construction, Equipments, etc
268(1)
27.4.2 Laboratory Staffs
268(1)
27.4.3 Organization Set-Up and System Protocol
268(1)
References
269(2)
28 Laboratory Safety and Laboratory Waste Disposal
271
28.1 Laboratory Waste Disposal
273(1)
28.2 Disinfectant Used in for the Contaminants
274(1)
References
275
Dr. Pranab Dey is a professor at the department of cytology and gynecologic pathology at the Postgraduate Institute of Medical Education and Research, Chandigarh. Professor Dey completed his MBBS at R. G. Kar Medical College and Hospital, Calcutta, M.D. (Pathology) at the Postgraduate Institute of Medical Education and Research, Chandigarh and his FRCPath (Cytopathology) at the Royal College of Pathologists, London. Professor Dey has conducted several research projects and has pioneered work on DNA flow cytometry, image morphometry, mono-layered cytology and cytomorphologic findings in various lesions in cytology smears. He has written numerous publications and is a member of various societies.
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