Manual of Medical Laboratory Techniques S Ramakrishnan, KN Sulochana
INDEX
Page numbers followed by f refer to figure and those followed by t refer to table
A
Abnormal RBC morphology 185f
Absolute eosinophil count 190
Acanthamoeba culture 335
Acanthocytosis 185
Acetic acid 247, 250
suspension 237
Acetoacetic acid 241, 249
Acetone 249
Acetoxylene 411
Acid
fast bacilli
culture 320
stain 412
phosphatase 65, 67
Activated partial thromboplastin time 208
Acute viral infections 264
Advantages of
formic acid decalcification 387
paraffin embedding 393
Agarose
concentration 147
gel electrophoresis 146
Alanine aminotransferase 60
Albumin 22
Alcian blue staining 405, 405f
Alcoholic formalin 400
Alizarin red-calcium complex 419
Amikacin 332
Amino acids 91
Amoxycillin 332
Analysis of CSF 259
Angiotensin converting enzyme 68
Anisocytosis 185
Antibiotic sensitivity 318
Antigen control tubes 362
Apple green birefringence 407f
Arginine 98
Ascorbic acid 246
in urine 240
Aspartate transaminase 58
Automatic tissue processor 389f
Avant genetic analyser 153f
Avidin biotin complex technique 423
B
Bacillus subtilis 316
Bacteria contain peroxidase 242
Barium chloride 252
Basic fuchsin 411
Bence Jones protein 257
Benedict's qualitative reagent 246
Benhold's method for amyloid 406
Benzidine 237
powder 237, 256
test 256
Beta hydroxyl butyric acid 241
Bile aesculin hydrolysis 315
Bilirubin 9, 241, 244
Bleaching
of melanin pigments 417
technique 418
Blood test 239
Boiling test 248
Borax solution 414
Brick red precipitate 246
Brucella agglutination test 360
Buffer solution 421
Buffered substrate 72, 76
Bunsen burner 246, 247, 258
C
Calcium 44
oxalate crystals 265
Calibration procedure 342
Carbohydrates 99
Carbonate buffer 83
Carboxylated sialomucins 405
Catalase test 308
Cefazolin 332
Cefotaxime 332
Ceftazidime 332
Cell count-using Neubauer chamber 261
Cerebrospinal fluid proteins 27
Ceruloplasmin 25
Chlamydia trachomatis 286, 337
Chloramphenicol 332
Chlorpromazine 241
Choice of fixative solution 382
Cholesterol 12
Chromatography 89
Chromogen 371
Chromosome abnormalities 127
Ciprofloxacin 332
Citrate utilization test 313
Classification of chromosomes 128
Clindamycin 332
Clot retraction 217
Clotting time 215
Coagulase test 309
Coating of slides 377, 395
Cocktail preparation for setting PCR 144t
Collection of
specimens 303, 306
stool sample 107
test sample 106
urine sample 106
Complete blood count 170
Concentration method 235
for detection of mycobacterium 274
Congo red method for amyloid 406
Conjunctival
papilloma 404f
scraping 294, 332
swab 332
Contact lens 332
Coombs' test 206, 207
Cornea 425
Corneal
button 332, 381, 387
scraping 299, 300, 332
stroma 407f
Counter stain 400
C-reactive protein 343
Creatinine 7
Cryostat knife 398
Culture methods 291
Cutting technique 399
Cyanmethemoglobin method 197
Cystine 102
crystals 265
Cytogenetic methods 116
D
Decalcifying fluid 387
Dehydration 389
Descemet's membrane staining pink 404f
Detection of
Bence Jones protein 257
hepatitis C virus antibodies 367
occult blood in stool 236
Diagnosis of ketonuria 250
Diluted serum 72
Direct
and indirect Coombs' test 206
bilirubin 11
Coombs' test 207
smear examination 234
Disposable blades 398
Disposal of materials 110
Distilled water 35, 67, 81
Donor corneoscleral rim 332
Down syndrome 133f
Drops of glacial acetic acid 257
Drying ovens 395
Durham's tube 311
E
Ehrlich's
aldehyde reagent 255
reagent 255
Electrophoresis 85, 200
buffer 148
of serum lipoproteins 86
of serum proteins 85
Electrophoretic techniques 145
Electrophoretogram 151
ELISA test 368
End-point of decalcification 388
Enterobacter aerogenes 315
Enterobacteriaceae 317
Enterococcus faecalis 315, 316
Eosin stock solution 401
Eosinophils 181
Eosin-phloxine working solution 401
Epinephrine 83
Epithelial cells 426f
Equilibrated phenol 140
Erythrocyte sedimentation rate 186
Erythromycin 332
Escherichia coli 311, 312
Ethanol solution 135
Ethidium bromide 148
Examination of malarial parasite 223
Examination of
microfilaria 226
stained smears 225, 228
External ocular specimens 332
Eye karyotyping 127
F
Femoral
tap 106
vein 106
Fermentation
of glucose 317
test 311
Ferric chloride 409, 410
Fibrinogen assay 218
Fixation 387, 422
time 381
Fluorescent antinuclear antibody test 376
Formaldehyde 383
Formamide wash solution 135
Formation of air bubbles 236
Formula for calculation of annealing temperature 145f
Fouchet's reagent 252
Fragile-X syndrome 127
Fructose 240
G
Gain of chromosome 127
Galactose 240
Gallego's solution 411
Gatifloxacin 332
Gene expression 162
General laboratory techniques 423
Genescan PCR conditions 158t
Genomic DNA extraction from blood 138
Gentamicin 332
Giemsa
banding 124
buffer 124, 280
stain 125, 279, 420
stock solution 125
working solution 125, 421
Glacial acetic acid 237, 256, 421
Glass
test tubes 249, 253, 255, 256
tube 237
Glucose
6-phosphate-dehydrogenase 73
tolerance test 4
Glucuronic acid 246
Glutaraldehyde 381, 383
Glutathione 39
peroxidase 79
Glycine 98
Glycoproteins 405
Goblet cells 404f
Gold chloride solution 414
Gomori's trichrome stain 408
Gram's
iodine 411
negative bacilli 317
stain 268
for bacteria 411
staining procedure 270
Granular
casts 265
dystrophy of corneal stroma 409f
Granulocytic leukocytes 239
Grocott's methenamine silver nitrate 413
Grossing
of biopsy specimens 386
technique 383
H
Harri's hematoxylin 400
Harvesting of chromosomes 121
HCl-potassium ferrocyanide working solution 418
Hematology 170
Hematoxylin 402f
Hemoglobin
electrophoresis 199
estimation 197
High
density lipoprotein 15
urinary protein 242
Homocysteine 30
Homocystinuria 104
Homogentisic acid 246
Howell Jolly bodies 185
Hyaline casts 265
Hydrogen peroxide 237
solution 257
Hydroxy butyric acid 249
Hypochromasia 185
I
Inborn errors of metabolism 99
Indirect Coombs' test 207
Indole test 314
Infected
red cell 226
suture 332
International System for Human Cytogenetic Nomenclature 132
Iron
and iron-binding capacity 48
buffer reagent 50
color reagent 50
Isochromosome 127
Ivy method 213
K
Ketone bodies 245, 249, 250
in urine 249
KOH-calcofluor white stain 271
L
Lactate dehydrogenase 70
Lactophenol cotton blue stain 283
Leishman
buffer 260, 262
stain 260, 262
Leukocytes 242, 244, 245, 265
Levey Jennings chart for SPC period 111f
Lid margin swab 297
Light microscope 260
Litmus paper 234
Low-density lipoprotein cholesterol 15
Lower level of gastrointestinal tract 234
Lymphocyte 181
M
Malignant melanoma choroid 385f
Mannitol
fermentation 312
motility test 312
Mantoux test 263, 264
Manual processing schedule for eyeball 391t
Manufacturer of urine strip 244
Maple syrup urine disease 101
Mature trophozoites 226
Mayer's/Harri's hematoxylin solution 406, 417
Measles 264
Metaphase chromosomes 121
Metaphosphoric acid 56
Methenamine 414
solution 414
Methicillin 320
Methionine 98
Microtome 394, 394f
Molecular cytogenetics 132
Morganella morganii 313
Moxifloxacin 332
Multiple acid fast bacilli staining 413f
Mumps 264
Myocilin gene exons 148f
N
Neubauer counting chamber 259, 260
Neutral buffered formalin 383
Neutrophils 181
Nigrosin stain 281
Nitrate reduction test 314
Nitric acid 387
Norfloxacin 332
Normal urine 248, 250
Novobiocin 320
Nuclear fast red solution 418
O
Ofloxacin 332
Ornithine 78
aminotransferase 77
Oxalic acid 418
Oxidase test 309
P
Packed cell volume 196
P-aminosalicyclic acid 241
Paraffin
embedding procedure 393
section cutting 394
wax
embedding 392
impregnation 390
Parasites, erythrocytes and pus cells 233
Pasteur pipette 248
Penicillin 332
Periodic acid and Schiff stain 403
Peripheral
blood lymphocyte culture 117
smear study 182
Perl's iron stain for hemosiderin pigments 418
Phenylalanine 98
Phenylenediamine 309
Phenylketones 241
Phenylketonuria 105
Phloxine
solution 400
stock solution 401
Phosphate solution 81
Phosphorus 46
Picking up sections 397
Picro acetone 411
Piperacillin 332
Plasma
amino acids 98
stain 407
Polio 264
Polyacrylamide gel electrophoresis 148
Polychromasia 185
Polymerase chain reaction 142
Porphobilinogen 241
Positive citrate control 314
Potassium
ferricyanide 66, 67
ferrocyanide stock solution 418
hydroxide 271
Preparation of
agarose gel 146
Coombs' control cells 207
fresh smear 227
glass slides 377
hemolysate 200
Kovac's reagent 315
nitrate reagent 314
oxidase reagent 309
smear 179, 278, 280
substrate 371
thin smear 225
Prometaphase chromosomes 121
Propylene glycol solution 417
Prostatic cancer 67
Protein 240
Proteus mirabilis 314
Prothrombin time 211
Protocol for
cycle sequencing reaction 153t
DNA extraction 140
setting RT reaction 163t
Providencia alcalifaciens 314
Pseudomonas aeruginosa 311, 314
Q
Qualitative
detection of
bile pigments by Fouchet's method 251
bile salts by Hay's test 252
free 256
identification of
protein in urine by heat and acetic acid method 247
reducing sugar in urine by Benedict's test 245
test 342
Quantification of
DNA 141
gene expression 162
R
RBC pipette 174f
Red cell agglutination 185
Reticulocyte count 192
Retinal membranes 381
Retinoblastoma 133f, 385f, 402f
Rheumatoid arthritis test 341
RNA
extraction 162
purification 163
Rothera's mixture 249
Rouleaux formation 185
Routine
paraffin procedures 422
staining 400
S
Salmonella typhi 314
Sanger dideoxy sequencing 151
Sartorius filter apparatus 168
Schistocytosis 185
Schuffner's dots 226
Screw-capped bottles 167
Sebaceous gland carcinoma 416f
Section of corneal button 407, 409f
Sectioning technique of globe 385f
Semi-quantitative
estimation 356
test 343, 345, 349, 352
Sensitivity of blood test 242
Separates
cell lines or clones 127
chromosome modal number 127
Serum
antistreptolysin ‘o’ titer 347
glutamic
oxaloacetic 58
pyruvic transaminase 60
Setting of lymphocyte culture 117
Sex chromosomes 127
Sickle cell preparation 231
Silver nitrate
solution 414
stock solution 414
working solution 414
Slide
agglutination method 203
coagulase for free coagulase 310
Sodium
azide 81
bisulfite 414
chloride mixture 388
hydroxide solution 406
nitroprusside
in test tube 249
solution 250
thiosulfate 414
solution 410
Sorensen's buffer 125
Spherocytosis 185
Squamous epithelial cell 265
Stained cellular components 260
Staining
method 322, 425
of blood film 180
techniques 268, 400
Staphylococcus aureus 309
Steatorrhea 234
Sterile test tubes 260
Stomatocytosis 185
Stool examination 233
Streptococcus pneumoniae 309
Sulfanilic acid 314
Sulfur powder 253
Sulfuric acid 35
Superoxide dismutase 82
Symbols and abbreviations used in karyotyping 127t
T
Tear drops 185
Temporal artery biopsy 410f
Test tubes 246, 248, 258
Testing for urobilinogen 255
Tetracycline 332
Thermometer 258
Thin with white flakes 234
Thiobarbituric acid reactive substances 37
Threonine 98
Throat swab 331
Tissue
floatation bath 396f
processing 389
Tobramycin 332
Total
bilirubin 11
erythrocyte count 173
proteins 20, 238
Touch-down PCR 144
Transaminase 58
Transitional epithelial cell 265
Treponema pallidum hemagglutination 353
Triacylglycerols 18
Trichrome staining of eyelid 408f
Triglycerides 18
Triple
phosphate 265
sugar iron agar test 313
Trophozoites 226
Tube agglutination method 203
Tuberculin syringe 263
Turk's fluid 260
Types of PCR 144
Tyrosine 98, 103
needles 265
U
Urea 6
Urease test 312
Uric acid 28, 246
crystals 265
Urinary tract infections 242
Urine 321, 332
deposit examination 265f
dropper 248
of menstruating females 244
of pregnant women 240
Urobilinogen 239, 241, 244, 255
V
Vacuum
impregnation 390
pump 119
Valine 98
van Gieson solution 410
Vancomycin 320, 332
Verhoeff's
elastic stain 409
working solution 410
iodine solution 409
Visualizing hybridization 137
Vitamin
A 51
C 54
E 53
von Ebner's hydrochloric acid 388
W
Washing procedure 136
Waxy casts 265
WBC
casts 265
pipette 174f
Wedge-type blood film preparation 182f
Weigert's iron hematoxylin 407
Western blotting technique 372
Widal test 357
Working
conjugate 373
diluent buffer 373
solution 412
of buffer 280
X
Xylene 391
Z
Ziehl-Neelsen stain 273, 277
×
Chapter Notes

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1
Manual of Medical Laboratory Techniques2
3
Manual of Medical Laboratory Techniques
Editors S Ramakrishnan MA PhD FAMS Professor Emeritus Department of Biochemistry & Cell Biology Vision Research Foundation Sankara Nethralaya, Chennai, India KN Sulochana PhD Director Department of Biochemistry & Cell Biology Vision Research Foundation Sankara Nethralaya, Chennai, India Foreword SS Badrinath
4
Jaypee Brothers Medical Publishers (P) Ltd.
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© 2012, Jaypee Brothers Medical Publishers
All rights reserved. No part of this book may be reproduced in any form or by any means without the prior permission of the publisher.
Inquiries for bulk sales may be solicited at: jaypee@jaypeebrothers.com
This book has been published in good faith that the contents provided by the contributors contained herein are original, and is intended for educational purposes only. While every effort is made to ensure accuracy of information, the publisher and the editors specifically disclaim any damage, liability, or loss incurred, directly or indirectly, from the use or application of any of the contents of this work. If not specifically stated, all figures and tables are courtesy of the editors. Where appropriate, the readers should consult with a specialist or contact the manufacturer of the drug or device.
Manual of Medical Laboratory Techniques
First Edition: 2012
9789350256343
Printed at
5
To
His Holiness Kanchi Maha Perriyaval with Our Sincere Pranams
6
7CONTRIBUTORS
8
9FOREWORD
I do not indulge in any language of exaggeration when I say that the book entitled Manual of Medical Laboratory Techniques released by the Medical and Vision Research Foundation and Vidyasagar Institute of Biomedical Technology and Science, Sankara Nethralaya is “All-in-one” in two respects—it covers all the relevant subjects, namely Clinical Biochemistry, Medical Microbiology, Hematology, Genetics and Molecular Biology and all the necessary diagnostic tests in each specialty. The analytical procedures form a large spectrum—from the common tests like blood glucose estimation and simple microscopy to highly-sophisticated techniques like High Performance Liquid Chromatography.
There are no two opinions in the fact that application of medical laboratory techniques goes a long way in the diagnosis and differential diagnosis of diseases. It is common knowledge that correct diagnosis is a ‘MUST’ to institute proper therapy and thereby alleviation of human suffering. As such, this book will help the medical fraternity to achieve the goal of ‘EFFECTIVE PATIENT CARE’. I have no doubt that the technical staffs involved in analytical work, scientists, doctors and researchers would find the information given in the book very useful. In addition, votaries of microbiology and biochemistry in science colleges would also be benefited, as such books are few and far between. It is worth having the book as a priced, lifetime possession of the concerned staffs and the students, and in the libraries of medical institutions, hospitals and science colleges.
The untiring efforts of Dr S Ramakrishnan in education and training to younger generation imparted in our institution, are most laudable.
The book is jointly authored by highly qualified and experienced teachers who are also researchers and technologists of Medical and Vision Research Foundation and Vidyasagar Institute of Biomedical Technology and are experts in the field. Hence the “last word” has been given by them in the performance of each test efficiently and with precision. I could imagine the hard work they would have put in preparing the text, in spite of their heavy academic and professional schedule. I hasten to congratulate them for their valuable contributions to the discipline of medical technology and thereby service to the patients.
I thank Jaypee Brothers Medical Publishers (P) Ltd., New Delhi, for having readily agreed to publish the book. I know they are second to themselves in the publication of a series of Medical books of value.10
As Chairman-Emeritus, I am happy to record that the ushering-in of the Manual of Medical Laboratory Techniques has got one more feather to adorn the crown of Sankara Nethralaya, the mother-organization of Medical Research Foundation and Vision Research Foundation and Vidyasagar Institute of Biomedical Technology and Science.
SS Badrinath
frcs(c) frcs (edin) dsc fams
Chairman-Emeritus, Sankara Nethralaya
Chennai, India
11PREFACE
“There is no wealth above the wealth of health.”
(Proverbs and wise sayings, Paul Vithayathil, 7th Edn, Vithayathil Publications, Cochin, India).
One can afford to lose wealth but not health which is a ‘must’ for happiness in life. No wonder ‘health for all’ is our goal. Strictly speaking, it should be the international goal too. For achieving this, diseases have to be erased from the Globe. It is impossible, as the cheapest commodity available at present appears to be only diseases! Hence, all attempts should be made to control and cure the diseases, to make the people happy—thanks to the medical and the technological professionals who strive and serve, day-in and day-out, devoting themselves to the care of the patients and alleviation of their sufferings.
Therapy and cure of a disease will be a success only if the diagnosis is correctly made. Application of Medical Laboratory Techniques is the sheet anchor for proper and precise diagnosis. Medical laboratory technologists belong to various specialties like Medical Microbiology, Hematology, Histopathology, Clinical Biochemistry, Human Genetics and Molecular Biology. Though the disciplines are different, there is unity in diversity, as the one and only mission of their votaries is to use, and, if possible, improve and innovate unfailing and sophisticated techniques in their respective fields in diagnosis, differential diagnosis, monitoring and if possible, cure of diseases of bewildering complexity—be they infectious or immunological, hematological or pathological, metabolic or genetic or just common.
The book Manual of Medical Laboratory Techniques is the essence of the subject jointly authored by highly qualified and experienced teachers, researchers and technologists of Medical and Vision Research Foundations, Vidyasagar Institute of Biomedical Technology and Science, Sankara Nethralaya, India, and is a presentation of diagnostic tests in their own specialties. Principles, methodologies, results, norms, interpretations, diseases concerned and bibliography have been given for the tests. Attractive illustrative diagrams are also given. Equal coverage has also been made for instrumentation. In short, it is all-in-one in medical laboratory techniques. A fact which deserves special mention is that a laboratory scientist can perform these tests without external guidance. The authors are confident that medical and technical institutions, hospitals, clinical laboratories, teachers, clinicians, technicians and students will find the book very useful and worthy of life-time preservation.12
The authors do not find suitable words to express their sincere thanks to Dr SS Badrinath, Chairman Emeritus, and Dr Bhaskaran, Chairman of Medical Research Foundation for their constant encouragement and Jaypee Brothers Medical Publishers (P) Ltd., New Delhi, for publishing the book. Thanks are also due to many colleagues who have helped in the manuscript preparation of the book, correction of proof and secretarial work. Our special thanks are due to Smt K Parvathy Devi of the Biochemistry Research Department for sustained coordination and giving the final shape to the book.
S Ramakrishnan
KN Sulochana