Ebook Concise book of medical laboratory technology - Methods and interpretations (2nd edition): Part 1

Concise Book of
Medical Laboratory Technology
Methods and Interpretations



Concise Book of
Medical Laboratory Technology
Methods and Interpretations
2nd Edition

Ramnik Sood MD (Path, Gold Medalist)
Consultant
Reem Medical and Diagnostic Center
Healthcare Mena Limited
Sharjah
United Arab Emirates

The Health Sciences Publisher
New Delhi | London | Philadelphia | Panama


Jaypee Brothers Medical Publishers (P) Ltd
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Phone: +91-11-43574357
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Email:

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© 2015, Ramnik Sood
The views and opinions expressed in this book are solely those of the original contributor(s)/author(s) and do not necessarily represent
those of editor(s) of the book.
All rights reserved. No part of this publication may be reproduced, stored or transmitted in any form or by any means, electronic,
mechanical, photocopying, recording or otherwise, without the prior permission in writing of the publishers.
All brand names and product names used in this book are trade names, service marks, trademarks or registered trademarks of their
respective owners. The publisher is not associated with any product or vendor mentioned in this book.
Medical knowledge and practice change constantly. This book is designed to provide accurate, authoritative information about the
subject matter in question. However, readers are advised to check the most current information available on procedures included and
check information from the manufacturer of each product to be administered, to verify the recommended dose, formula, method and
duration of administration, adverse effects and contraindications. It is the responsibility of the practitioner to take all appropriate safety
precautions. Neither the publisher nor the author(s)/editor(s) assume any liability for any injury and/or damage to persons or property
arising from or relate♥d to use of material in this book.
This book is sold on the understanding that the publisher is not engaged in providing professional medical services. If such advice or
services are required, the services of a competent medical professional should be sought.
Every effort has been made where necessary to contact holders of copyright to obtain permission to reproduce copyright material. If any
have been inadvertently overlooked, the publisher will be pleased to make the necessary arrangements at the first opportunity.
Inquiries for bulk sales may be solicited at:
Concise Book of Medical Laboratory Technology: Methods and Interpretations
First Edition: 2009
Second Edition:  2015
ISBN 978-93-5152-333-8
Printed at

Dedicated to
the Readers



Preface to the Second Edition
I authored an exhaustive book entitled “Medical Laboratory Technology – Methods and Interpretations” that hit the stands
in mid-eighties in the previous century and is now running in its 6th edition. This was the first such book in the subcontinent and was much appreciated and used by technologists and pathologists alike. The book has seen testimonies in courts
and has been appreciated in the west too. However, in our subcontinent, I was requested by many technologists that they
wanted a little younger sister of the book popularly known as MLT authored by me.
And so was born the Concise Book of Medical Laboratory Technology: Methods and Interpretations. The book essentially
covers everything presented in MLT-6 but in an abridged/shortened and easy-to-digest format. The book is presented in
a flowing noninterrupted format and not in a cumbersome experiment-wise cascading flow. There is no break in the style
that runs smoothly and is easier to absorb and assimilate. As experiments are a part of any course, more stress has been
laid out in the book to understand the intricacies of relevant theories and even troubleshooting all experiments that you
would conduct during the course of your study. As I have written multiple modules for many Universities in India, I did not
have to think for too long to devise a style and format for this book. You will find everything from ESR to PCR and you will
find foam test for bile pigments as also complete automation in urinalysis. You will find basic biochemistry as also detailed
cytogenetics. So whatever be your course or query, you will find it within the covers of this short but sweet book now going
into its second edition.
The book is designed for you not to mug but to understand and elicit the answers from the book of all questions in your
mind. I am aware that this book is used by most of your teachers and tutors too.
Nothing wrong that you are holding now. It will help you all your life!

Ramnik Sood



Preface to the First Edition
The first book on Medical Laboratory Technology from the house of Jaypee’s came out in 1985 and has been the best seller

in its class till date. The title “Medical Laboratory Technology – Methods and Interpretations” has seen 6 editions. The latest
one hit the stands in January this year and was released in two volumes. It is a four-color book and has over 1670 pages. The
reader base of all editions of our vastly popular title “MLT” has been upcoming laboratorians, undergraduate and postgraduate
medical students. It was requested by a few institutes to produce a little smaller version of the two-volume set that would
be suitable for the upcoming Laboratory Technology students. So here it is! It is exhaustive yet precise and concise too.
The book will help you to appreciate things as they appear in real life under the microscope and otherwise. All current
technologies find a mention within the covers of this book. Gone are the days when we had to prepare reagents first thing
in the morning (or the days usage); therefore, the current trend of consuming ready-to-use reagents/kits is followed here
to make your job and understanding simpler. So, what is available in the market for all investigations, is what is presented
here. The Tulip Group has very kindly given us the rights to reproduce the text related to all their kits and reagents in this
book. Latest instruments are not forgotten too.
Most important—
Parasitology section is presented in ample detail as it is relevant to all the developing nations.
Quality control/assurance is mentioned in appropriate details. Working is not enough. Working properly and producing
nothing but the most accurate reports are the order of the day. This book will not fail you there. Follow the recommendations to the hilt and you would be running the most accurate laboratory available anywhere.
Should problems arise! The book has “Troubleshooting” section for every possible test mentioned inside. If this happens – then what! If that happens – then what! You will find all answers.
From ESR to PCR – you will find everything.
The book is based on most syllabi as applicable to most institutes in India and elsewhere internationally.
All necessary care has been taken to weed out any discrepancies/typographical errors at the time of going to press.
However, if anything has remained inadvertently, the publishers/author do not take any responsibility for the same in any
manner whatsoever.
Learning can be enjoyable experience, flip a few pages to experience that.

Ramnik Sood



Contents
Chapter  1. Laboratory

Laboratory Set-up  3
Code of Conduct for Medical Laboratory
Personnel  6
Accidents  6
Accidents in the Laboratory  10
Universal Work Precautions (Uwp) for
Laboratory Personnel (Especially in
Relation to Hiv Transmission)  15
Medicolegal Aspects of Clinical Practice  17
Laboratory Instruments  17

1

29

Chapter  3. SI Units

41

Chapter  4. Fundamental Chemistry

52

Chapter  5. Urine Analysis

56

Methods Commonly Used for Sterilization  29
Modern Day Disinfection  34

Indicators  52
Solutes, Solvents and Solutions  52
Periodic Table of Elements  54
Composition of Urine  56
Gross Examination of Urine  57
Chemical Examination of Urine  60
Multiple Reagent Strips for Urinalysis  71
Automation in Urinalysis  78
Special Urine Tests  82
Microscopy of the Urinary Sediment  92

Chapter   6. Renal Function and its
Evaluation104
Renal Physiology in Brief  104
Functions of the Kidney  104
Concentration: Dilution Tests  105
Phenol Red Test  105
Clearance Tests  106
Principles of Precise Tests of Renal
Function  107
Maximal Tubular Capacity (Tm)  108

112

Chapter  8. Medical Parasitology

124

Chapter  9. Clinical Hematology

205

Chapter  10. Clinical Hematology:
Bleeding Disorders

272

Specimen Collection  112
Inspection of Feces  113
Medical Parasites  124
Intestinal Protozoa of Man  124
Laboratory Examination for
Parasites  201

Chapter  2. Sterilization

Liter  41
Gram  41
Mole (Mol)  41
International Unit (U)  42
Conversion Factors Between Conventional
and System International Units (Siu)  42

Chapter  7. Stool Examination

Ways of Obtaining Blood  205
Anticoagulants  206
Blood Collection System  208
Hemoglobin  210
Anemia  211

Hematocrit/Packed Cell Volume (Pcv)  212
Blood Cell Counts  213
Erythrocyte Indices  216
Complete Blood Count (Cbc)  217
Erythrocyte Sedimentation Rate (Esr)  220
Blood Film Examination  222
Rapid Diagnostics  225
Development of Blood Cells and
Sites of Blood Formation  227
Morphological Types of Red Blood Cells  234
Qualitative Assessment of G6pd
Deficiency  240
Examination of Fetal Hemoglobin  244
Laboratory Diagnosis of Disorders Related to
Rbcs  248
Thalassemias (Reduced Synthesis Rate)  256
Normal White Cell Values and Physiological
Variations  259
White Blood Cells  263
Quality Control in Hematology  270

Platelets, Coagulation and Bleeding Disorders:
Laboratory Investigations  272
Laboratory Diagnosis of Platelet Disorders  272
Quality Assurance for Routine Hemostasis
Laboratory  278
Buffered 3.2% Citrate Solution (Profact)  280
Prothrombin Time (Quick One-Stage Method)
Liquiplastin®  283
Sensitive Thromboplastin Reagent for

Prothrombin Time (Pt)
Determination (Isi = 1.0) Uniplastin®  285


xii

Concise Book of Medical Laboratory Technology: Methods and Interpretations
Thromboplastin Reagent for
Prothrombin Time (Pt)
Determination, Lyoplastin®
(Lyophilized Reagent, Isi = 1.0)  287
Aptt/Pttk Cephaloplastin Reagent for Partial
Thromboplastin Time (Aptt)
Determination Using Ellagic Acid as
Activator Liquicelin-E®  293
Normal and Abnormal Control Plasmas for
Coagulation Assays Plasmatrol H-I/Ii®  296
Fibroscreen Thrombin Time Test for Qualitative
Estimation of Fibrinogen Fibroscreen®  297
Fibrinogen Estimation-Quantitative
Fibroquant, Reagent for Quantitative
Estimation of Fibrinogen  299
Fibrinolytic Activity  301
Fdps A Qualitative and Semiquantitative Latex
Slide Test for Detecting Cross
Linked Fibrin Degradation Products in
Human Plasma X-L Fdp  302
Laboratory Diagnosis of Coagulation
Disorders  304
Automation in Coagulation Analysis  305

Troubleshooting  309
Prothrombin Time  310
Dptt/Pttk  312
Fibrinogen Estimation  314

Chapter  11. Blood Banking
(Immunohematology)317

Blood Group Antibodies  317
Anti-A, Anti-B, Anti-Ab Blood Grouping
Antisera for Slide and Tube Tests  320
Anti-A, Anti-B, Anti-Ab Monoclonal }|Blood
Grouping Antibodies for Slide and Tube
Tests  321
Anti-A1 Lectin Dolichos Biflorus
Lectin for Slide and Tube Tests  322
Anti-H Lectin Ulex Europaeus Lectin
for Slide and Tube Tests  323
Physiological Saline Solution for Serological
Applications (Sodium Chloride 0.9%
W/V)  325
Bovine Serum Albumin 22% Solution
for Serological Applications  325
Concentrated Iso-Osmotic Phosphate Buffered
Saline for Serological Applications  327
Red Cell Preserving Solution for Serological
Applications  328
Abo Grouping  329
Rh Blood Group System  331
Anti-D (Rho) Human (Igg) Polyclonal Blood

Typing Antibodies for Slide
and Modified Tube Tests  333
Anti-D (Rho) (Igm) Monoclonal Blood Typing
Antibodies for Slide and Tube Tests  335
Anti-D (Rho) (Igg) Monoclonal Blood Typing
Antibodies for Slide and Modified Tube
Tests  336

Anti-D (Rho) (Igm + Igg) Monoclonal Blood
Typing Antibodies for Slide and Tube
Tests  338
Anti-human Igg Monospecific Coomb’s
Reagent for Direct and Indirect
Anti­globulin Test  353
Anti-human Globulin Reagent for Direct and
Indirect Antiglobulin Tests  356
Preparing Coomb’s Control Cells Agtrol®  358
Low Ionic Salt Solution for Serological
Applications  359
Stabilized, Activated Papain Enzyme Solution
for Serological Applications  361
Blood Transfusion  363
Trouble Shooting  372

Chapter  12. Cerebrospinal and Other
Body Fluids

382

Chapter  13. Semen Analysis

398

Chapter  14. Sputum Examination

405

Chapter  15. Pregnancy Tests

411

Chapter  16. Examination of
Gastrointestinal Contents

425

Cerebrospinal Fluid  382
Synovial Fluid (Sf)  388
Pleural Fluid  389
Pericardial Fluid (Pf)  391
Peritoneal Fluid  392
Amniocentesis and Amniotic Fluid
Analysis, Diagnostic  394
Semen Analysis  398

Sputum  405
Common Respiratory Disorders  406
Bioassays  411
Immunologic Methods  412
Slide Test for Pregnancy  412

Slide Test for Pregnancy  414
Elisa Pregnancy Test  416
Dipstick Ict Pregnancy Test  416
Device Ict Pregnancy Test  417
Dipstick Ict, Urine/Serum
Pregnancy Test  418
Device Ict Urine/Serum
Pregnancy Test  419
Troubleshooting  421
Rapid Formats  423

Normal Saliva—Constituents  425
Gastric Juice  425
Examination of Duodenal Contents  430
Composition of Bile  430
Pancreatic Function Tests  430
Sweat Electrolytes Pilocarpine
Iontophoresis  432


Contents
Chapter  17. Diabetes Mellitus:
Laboratory Diagnosis

434

Chapter  18. Liver Function Tests

454

Chapter  19. Clinical Chemistry

461

Chapter 20. Enzymology

519

Diabetes Mellitus  434
Glycosylated Hemoglobin Kit (Ion Exchange
Resin Method) for the Quantitative
Determination of Glycohemoglobin in
Blood (for in Vitro Diagnostic use Only)  443
Rapid Diagnostics  448
Tests of Excretion by the Liver  454
Evaluation of Synthesis in Liver  457
Evaluation of Enzyme Activity  458
Suggested Liver Function Tests  458
Colorimetry  461
Photometer  462
Clinical Chemistry  465
Total Proteins  478
Serum Albumin  479
Serum Cholesterol  481
Hdl Cholesterol  484
Blood Glucose  490
Uric Acid  492
Calcium  493
Phosphorus  497
Chloride  500

Serum Iron and Tibc  501
Trace Elements  503
Zinc  503
Zinc (Colorimetric Method)  503
Copper  504
Magnesium  506
Automation in Clinical Chemistry  508
Principles of Quality Assurance and
Standards for Clinical Chemistry  514

Alpha-amylase  519
Lipase  520
Phosphatases  522
Transaminases  530
Gamma-glutamyl Transpeptidase (Ggtp)
Blood  536
Lactic Dehydrogenase  538
Automation in Clinical Chemistry: Random
Access Autoanalyzer  546

Chapter  21. Blood Gases and
Electrolytes548
Blood Gases  548
Automation in Blood Gas Analysis  556
Avl Compact 2 Blood Gas Analyzer  557
Electrolyte Analysis by Flamephotometer  558
Rapid Diagnostics in Electrolyte Analysis  561

Chapter 22. Serology/Immunology
Basic Immunology  563

Technologies  568

563

xiii

Enzyme Immunoassay  572
Chemiluminescence: The Technology  585
Polymerase Chain Reaction  587
Ria  590
Liquid Handling Systems  591
Streptavidin-Biotin Systems  594
Representative Elisa/Clia Techniques  595
Examples of Detailed Elisa
Methods  597
Tests for Syphilis  608
Modified Vdrl Reagent Trepolipin®  610
Toluidine Red Unheated Serum Test for Rapid
Serodiagnosis of Syphilis Redgen®  613
Latex Slide Test for Vdrl Syphfinal  615
Rapid Plasma Reagin (Rpr) Card Test/Carbon
Antigen for Syphilis Testing (Carbogen)  618
One-step Test for Syphilis: Dipstick
Syphicheck®  621
One-step Test for Syphilis (Device)
Syphicheck  622
Third Generation Double Antigen Sandwich
Enzyme-linked Immunosorbent Assay
(Elisa) for the Detection of Antibodies to
Treponema Pallidum in Human

Serum or Plasma Trepolisa 3.0  624
Tests for Typhoid/Enteric Fever Widal
Antigen Set/Antigens for Tube Tests
(Typhochek)  624
Widal Antigen Set/Antigens for Slide
and Tube Tests (Tydal)®  627
Reduced Widal Antigen Set: O and H
for Tube Tests (Vital Widal)  630
Positive Control for Widal Test  631
Rapid Test for Detection Igm Antibodies to
S. typhi in Serum/Plasma/Whole Blood
(Device) Enterocheck – Wb  632
Slide and Tube Test for Detection of Antibodies
to Brucella Abortus/Melitensis Brucel
A/M  635
Slide Screening Test for Brucella Antibodies
(Brucel-Rb)®  636
Brucellosis Positive Control  638
Rapid Test for Igm and Igg Antibodies
to Dengue Virus: Dengue Fever
(Denguecheck-Wb) (Device)  639
Test for Infectious Mononucleosis
(Immutex)  643
Rapid Test for Igm Antibodies to Leptospira:
Leptospirosis (Leptochek-Wb) (Device)  645
Rapid Test for Malaria Pan/Pv/Pf
(Paramax-3®) (Device)  647
Slide Test for C-reactive Protein (Rhelax
Crp)  650
Slide Test for Antistreptolysin O (Rhelax

Aso®)  653
Slide Test for Rheumatoid Factors
(Rhelax Rf)  656
Slide Test for Anti-deoxyribonucleoprotein
(Rhelax Sle)  659


xiv

Concise Book of Medical Laboratory Technology: Methods and Interpretations
Australia Antigen Hbsag (Virutex Hbsag)  662
One-Step Test for Hbsag Virucheck Device  664
Hcv Flavicheck Device  665
Toxoplasma Infections  668
Rapid Immunoconcentration
Test for Hiv-1 and Hiv-2
Antibodies Flow through
Method Retroquick-Hiv  669
Rapid Test for Simultaneous/Differential
Detection of total Antibodies to Hiv1 and Hiv-2 in Human Serum/Plasma
Retroscreen  671
Tuberculosis  673
Rapid Test for Detection of Antibodies
to Mycobacterium tuberculosis
(Device) Serocheck-Mtb  676
Tb Igg, Iga, Igm Ab, Mfd Anda  678
Tumor Markers  679
Tumor Markers Standard Methodologies Avail­
able on Elisa and Clia, as on Ria too  683
Ca 15-3 (Carcinogenic Antigen 15-3)  683

Ca 19-9 (Carbohydrate Ag 19-9, Gicam
Gastrointestinal Cancer Antigen)
Blood Mfd: Can Ag,  683
Ca 242 Mfd: Can Ag,  684
Ca 125 (Cancer Antigen 125) Mfd:
Monobind  684
Carcinoembryonic Antigen (Cea) Mfd:
Monobind  685
Prostate-specific Antigen (Psa) Total
Prostate Specific Antigen (Tpsa) Elisa,
Mfd: Monobind  685
Prostatic Acid Phosphates (Pap), Blood
Method: Biochemical Analysis  686
Elisa Troubleshooting Aspects  686
Technical Tips  690

Chapter  23.Diagnostic Immunology

693

Qualitative Determination of Plasma Proteins
by Immunoprecipitation  693
Fundamental Quantitative Considerations  698
Turbidimetry  699
An Example of Turbidimetric
Immunoassay  718
C-reactive Protein  720
Turbidimetric Immunoassay for Determination
of C-reactive Protein  722
Turbidimetric Immunoassay for Ultrasensitive

Determination of C-Reactive Protein  723
Turbidimetric Immunoassay for Determination
of Antistreptolysin ‘O’ in Human Serum  724
Turbidimetric Immunoassay for Determination
of Microalbuminuria  724
Immunoglobulins (Ig)  724
Turbidimetric Immunoassay for Estimation of
Immunoglobulin
Iga in Human Serum  725
Turbidimetric Immunoassay for Estimation of
Immunoglobulin Igg in Human Serum  726

Turbidimetric Immunoassay for Estimation of
Immunoglobulin Igm in Human Serum  726
Turbidimetric Immunoassay for Estimation of
Complement C3 in Human Serum  726
Turbidimetric Immunoassay for Estimation of
Complement C4 in Human Serum  727
Turbidimetric Immunoassay for Estimation of
Antithrombin Iii in Human Serum  727
Quantitative Immunoturbidimetric
Assay for Estimation of Fibrinogen  727
Turbidimetric Immunoassay for Estimation of
Lipoprotein (A) in Human Serum  727
Quantitative Turbidimetric
Immunoassay for Estimation of
Apolipoprotein A-I  728
Quantitative Turbidimetric
Immunoassay for Estimation
of Apolipoprotein B  728

Automation in Turbidimetry  729

Chapter  24. The Endocrine System

731

Pituitary Gland  731
Anterior Lobe: Growth Hormone (Gh)  732
Method of Evaluation: StreptavidinBiotin Elisa  733
Corticotropin (Acth)  735
Other Anterior Pituitary Hormones  735
Intermediate Lobe (Pars Intermedia)  736
Posterior Pituitary (Neurohypophysis)  736
Disorders of the Pituitary System  738
Hypothalamus  738
Adrenal (Suprarenal) Gland  738
Mineralocorticoids  738
Glucocorticoids  739
Adrenal Medulla  742
Thyroid  742
Calcitonin  753
Parathyroid  754
Parathyroid Hormone (Intact) Elisa  756
Pancreas  757
Testes  758
Ovary  758
Pineal Gland  759
Hormones and Fertility  759
Male Fertility  760
Female Fertility  763

Algorithm for Evaluating Amenorrhea,
Immunoassays for Lh, Fsh and Prl  768
Adrenal Cortex  775
Adrenal Medulla  777
Testes  779
Steroids  781
17-Β-Estradiol  781
Dheas (Dehydroepiandrosterone Sulfate)  782
∆4-Androstenedione  782
Progesterone  782
17-Alpha-hydroxyprogesterone  783
Total Tri-iodothyronine (T3)  783
Cia™ Insulin (Chemiluminescence
Immunoassay)  788


xv

Contents
Chapter 25. Histopathology

Preparation of Tissues  791
Routine Staining Procedures  794
Some Staining Techniques in Detail  799
Automation in Histopathology  805

Chapter 26. Cytology

791

808

Papanicolaou Method of Staining
Smears (Modified)  808
Fnac (Fine-Needle Aspiration Cytology)  809
Smearing Techniques  812
Requirements for Laboratory Set Up  812
Immunoperoxidase Staining for
Cyto and Histopathology  817
Automation in Cytology  818

Chapter  27. Microbiology and
Bacteriology819
Classification  819
Culture  825
Ready to Pour, Sterilized Pouched Media
for Microbiological Applications
Instaprep  828
Easybact  833
General Instructions for Microbiology  837
Gram-positive Cocci  837
Gram-negative Cocci  839
Anaerobic Spore Bearing Bacilli  840
Aerobic Spore Forming Bacilli  842
Gram-positive Bacilli  842
Mycobacteria  843
Overview of M. tuberculosis: Diagnostic
Approach, Afb Staining, Culture
and Sensitivity  845
Mucolytic, Disinfectant, Specimen

Pretreatment and Buffering System
for Afb Staining and Culture  848
Rapid Two Step Cold Afb Stain  851
Ready to use Lj Solid Medium for
Mycobacterium tuberculosis Isolation  855
Combipack of Solid and Liquid Medium for
Mycobacterium tuberculosis Isolation  857
Primary/Secondary Drug Containing
Lowenstein-Jensen Media Panel Mtb
Sensitivity Tests  861
In Determination of Adenosine Deaminase
Activity in Serum, Plasma and Biological
Fluids  863
Gram-negative Bacilli  865
Spirochetes  871
Gram Stainer  872
Quality Assurance in Bacteriology  872
Color Atlas—Media and Colonies  873

Chapter 28. Mycology

883

Intermediate Superficial Deep Mycoses  884
Deep or Systemic Mycoses  884
Fungi Usually Present as Contami­nants but
Which Rarely Cause Disease—Usually in

Patients Chronically Ill from Other
Diseases  885

Mycological Methods  885

Chapter  29. Diagnostic Skin Test

886

Chapter 30. Cytogenetics

891

Chapter  31. World’s Latest and Best
Technologies by Roche

896

Technique of Skin Tests  886
Immunologic Basic for Skin Tests  886
Common Skin Tests  887
Immediate Reaction Type of Skin Tests  887
Delayed Reaction Type of Skin Tests  889
Blood Lymphocyte Culture  891
Karyotyping  893
G and Q Bandings  894
Importance of Chromosomal Studies  894
Barr Body Analysis and Buccal Smear for
Staining of Sex Chromatin Mass  895

Business Areas  896
Revaluating Diagnostics  896
Testing Efficiency and Medical Value  896

Effective Management of Infectious
Diseases  898
Cobas® Modular Platform  900
Your Benefit  900
Cobas® 8000 Modular Analyzer Series  901
Cobas® 8000 Modular Analyzer Series  902
Cobas® 6000 Analyzer Series  903
Cobas® 4000 Analyzer Series  906
Cobas C 311 Analyzer  906
Cobas E 411 Analyzer  906
Cobas C 111 Analyzer  907
Cobas Integra® 400 Plus  908
Cobas P 312 Pre-analytical System  910
Cobas P 512 Pre-analytical System  910
Cobas P 612 Pre-analytical System  911
Cobas P 501 and Cobas P 701
Modular® Pre-analytics Evo  912
Cobas® Connection Modules (Ccm)  913
Cobas® 8100 Automated Workflow Series  914
Cobas® It Solutions  916
Cobas® Middleware Solution  917
Cobas® Laboratory Information System  918
Cobas® Infinity It Solutions  920
Overview of Serum Work Area Tests  921
Ecl—Unique Immunoassay Technology  925
Technology for Homogeneous
Immunoassay Detection  926
Elecsys® Hbsag Ii Quant  926
Elecsys® Hiv Combi Pt 4th Generation (Ag+Ab
Test)  927

The Syphilis Assays  929
Elecsys® Syphilis Immunoassay  929
Elecsys® Torch Panel  930
Elecsys® Troponin T High Sensitive
(Tnt Hs)  931


xvi

Concise Book of Medical Laboratory Technology: Methods and Interpretations
Key Benefit: Earlier Diagnosis of Ami  932
Elecsys® Nt-proPnB  932
Elecsys® Tumor Marker Portfolio  933
Elecsys® He4  935
Elecsys® Pro-grp  936
Elecsys® Calcitonin  937
Elecsys® Anti-Tshr  938
Elecsys® Tg Ii  939
Tina-Quant® Lipoprotein (A) Gen 2 Test  940
Tina-Quant® Immunoglobulin A and M
Csf  941
Tina-Quant® Hemoglobin A1c  942
Tina-Quant® Cystatin C Gen 2  942
Elecsys® Preeclampsia  943
Elecsys® Vitamin D Total  945
Elecsys® Il-6, Pct and Tina-Quant® Crp  945
Elecsys® Tacrolimus and Cyclosporine  947
Hemostasis Testing  949
Multiplate® Analyzer  949
Urinalysis  951

Urinalysis From Roche  951
Combur-Test® Strip  951
Urisys 1100® Analyzer  952
Cobas U 411 Urine Analyzer  953
Cobas® 6500 Urine Analyzer Series*  954
Point-of-Care Testing  956
Cobas Poc it Solution  958
Cobas bge Link Software  960
Cobas B 121 System  961
Cobas B 221 System  962
Cobas B 123 Poc System  963
Accu-Chek® Inform Ii System  964
Accu-Chek® Safe-T-Pro Plus  965
Cobas H 232 Poc System  966
Roche Cardiac® Trop T Sensitive Test  967
Coaguchek® Xs System  968
Coaguchek Xs Plus System  969
Accutrend® Plus System  970
Reflotron® Plus System  971
Cobas B 101 System  972
Molecular Diagnostics  974
Solutions from Roche for Molecular
Diagnostics  974

Cobas P 630 Instrument  977
Cobas® Ampliprep Instrument  978
Cobas® Taqman® Analyzer and Cobas®
Taqman® 48 Analyzer  979
Cobas® Ampliprep/Cobas® Taqman® Hcv
Qualitative and Quantitative Tests, V2.0  980

Cobas® Taqman® Mtb Test  981
Cobas P 480 Instrument  982
Cobas® 4800 System V2.0  983
The Cobas® Hpv Test  985
The Cobas® Oncology Tests  985
Cobas® Mrsa/Sa Test  987
Cobas® Cdiff Test  987
Cobas® Hsv 1 and 2 Test  988
Cobas S 201 System  988
Lightcycler® Systems  990
Lightcycler® 2.0 Instrument  991
Test Kits, Validated for Ivd  992
Lightcycler® Septifast Test  992
Lightcycler® Mrsa Advanced Test  993
Magna Pure Systems  993
Symphony System  996
Benchmark Special Stains  997
Primary Antibodies  999
Ihc Detection  1001
Breast Cancer Diagnostics  1002
Prostate Cancer Diagnostics  1003
Hematopathology  1004
Colorectal Diagnostics  1004
Lung Cancer Diagnostic Solutions  1005
Benchmark Ihc/Ish Platform  1007
Benchmark System Features  1007
Digital Pathology  1008
Vantage Workflow Solution  1009
Consultancy Services  1011
Sequencing Solutions  1013

Genome Sequencer Flx+ System  1013
Gs Junior System  1014
Nimblegen Sequence Capture  1015
Roche Dialog  1017

Appendix1019
Index1033


CHAPTER

1

Laboratory
INTRODUCTION
The definition of health includes a state of complete and
perfect physical, mental, social and spiritual well-being
and not just the absence of disease or infirmity and good
health is a fundamental right of every living human
being on earth. However, modern world, though, has to
an extent eliminated infectious diseases. But the focus
has now shifted to lifestyle diseases. Pollution of every
nature too has taken its toll. About half a century back,
the predominant diseases used to be infective ones but
now you may find individuals in mid-twenties waiting
for their turn for open heart surgeries. Also, modern
medicine has increased the longevity of life accompanied
by attendant geriatric diseases like Alzheimer’s disease
and malignancies. The polluted and toxic world has
not spared the fetuses in utero and neonates. A new

face of disease has emerged, diseases like HIV-AIDS
and severe acute respiratory syndrome (SARS), are new
entrants in the long list of infective diseases. We may
have eradicated smallpox but tuberculosis and malaria
have raised their heads with a vengeance. So, do what
you might. Some forms of disease, mild or severe will
strike every human being living. On getting sick, the
patient first comes in contact with a clinician—medical
or surgical. The clinician gives a patient hearing (if the
patient is conscious) to his problems and symptoms and
also takes note of various signs, which he sees or elicits.
Sometimes, he may immediately arrive at a diagnosis
and may under emergency circumstances institute
treatment at first instances. In most cases, however, he
will have a differential diagnosis in mind and to arrive
at a specific diagnosis he usually orders for a battery of
tests.

Various means of diagnosis are available.
1.Most important: Clinical laboratory tests which include
any tissue or fluid obtained from the body.
2.Imaging sciences: X-rays, ultrasound, color Doppler,
computerized axial tomography (CAT) scan, magnetic
resonance imaging (MRI) scan and the latest positron
emission tomography (PET) scan.
3.Electrical signal processing techniques: ECG, EMG, EEG
and nerve transmission techniques, etc.
4.Direct visualization techniques: With the availability
of fiberoptic-based technologies, the clinician is
now capable of passing small tubes (called scopes)

through natural passage ways of the human body
(without actually surgically opening up the part),
e.g. gastroscopy, cystoscopy, etc. These techniques,
eventually culminate in taking small tissue samples
(biopsies) which are sent to histopathology laboratories.
So, whenever, any sample from a human body is taken
(either voided naturally or obtained by the clinician or the
laboratorian), it is referred to the clinical laboratory for
investigation. On receipt of a report from the laboratorian,
the clinician, then, makes up his mind and starts a
unidirectional or specific treatment against the disease
thus diagnosed. It would not be wrong to designate medical
laboratory personnel as the backbone of the clinicians. But,
for these technologists, the clinicians would forever grope
in the dark. Gone are the days when diabetes mellitus was
presented with the classical triad of symptoms—increased
thirst, hunger and urination; likewise, typhoid seldom
presents with a step-ladder pattern fever. Blood testing
is absolutely mandatory, to know that they exist, their
severity and eventually, after treatment; to know that they
are under control or cured. Investigations are diagnostic as
well as prognostic tools.


2

Concise Book of Medical Laboratory Technology: Methods and Interpretations

Clinical laboratory investigations nowadays are being
utilized as future predictors. On getting warning signals,

one can take necessary corrective measures (lifestyle and/
or dietary) and can prevent diseases from striking or at
least deferring or postponing their arrival.
HELP! AND YOU DID AND YOU ALWAYS WILL. When
a clinician is lost, you shall show him the way in the best
possible way, you lead him to a diagnosis and let him
do his job thereafter. He may come back to you later to
determine that his efforts have been fruitful.
The following pages within the covers of this book will show
you the right path on how to be an excellent laboratorian.
Do your best in serving mankind. As you yourself may be
a patient tomorrow. This book shall also serve you well by
providing interpretation of the results obtained by you.
This book shall be true to its title “Concise Book of Medical
Laboratory Technology: Methods and Interpretations”.
While physiology is the study of essentially normal
structures and functions of a body, pathology deals with
the study of a diseased organ or system of the body, its abnormal functions, their mode of origin, their progress to
recovery or otherwise. All these studies come under the
ambit of a clinical pathology laboratory. A clinical laboratory has further sub-branches such as: hematology, biochemistry, seroimmunology, microbiology, cytogenetics,
histopathology, cytopathology, blood banking and last but
not least—clinical microscopy.

A clinical laboratory can be manned by a qualified
doctor specializing in clinical pathology, biochemistry,
immunology, blood banking, histopathology, cytopathology,
hematology, microbiology or cytogenetics. The pathologist
is usually assisted by laboratory technicians or technologists
(they are also qualified for the job) and lastly the cleaning
and documentation staff. Only by collective efforts of

the individuals mentioned above, a proper report can be
generated. Be grateful to the clinician for having faith in you
and give back nothing except an accurate and correct timely
report. A delayed report may at times be too late. The patient
may have lost his life by then. A timely correct report is the
essence of running a good laboratory.
The cycle of health-disease with all intermediaries is
given in Figure 1.1. Just as there are primary, secondary
and tertiary health centers, there are also the primary,
secondary and tertiary laboratories too. In India, there are
no specific guidelines as to what or how much they can
do and overlapping can occur. A superior laboratory may
perform all functions of an inferior laboratory too.

Primary Laboratory
In rural setups, for instance, a primary laboratory may
provide only the basic investigations. These investigations
are simple to perform and do not involve expensive
machinery usage. Such laboratories are also attached to

FIG. 1.1: Health-disease-health cycle


Laboratory
physician chambers nowadays, so that clinicians may
obtain basic inputs right in their own premises. These
primary laboratories may provide the following simple
investigations:
¾¾ Hemograms (hemoglobin estimation, total and
differential counts, erythrocyte sedimentation rate and

packed cell volume with basic peripheral smear study
including the reporting of hemoparasites)
¾¾ Routine and microscopic studies of urine and stool.
Routine examination also entails chemical examin­
ation either by laborious and time-consuming old
chemical methods or by new generation dipstick
tests. These may include tests for glucose, bilirubin,
ketones, hemoglobin, leukocytes, pH, nitrites, protein,
urobilinogen and specific gravity in case of urine. For
stool samples, reducing substances, pH and occult
blood may be performed. Basic spot/latex/device tests
(e.g. pregnancy test) may be conducted.

Secondary Laboratory
These are laboratories that assist a clinician to confirm a
clinical suspicion or establish a diagnosis. Therapy and
prognosis monitoring can also be provided from these
laboratories. Such laboratories are staffed by qualified
personnel who are trained and experienced to perform the
tests. They also have a perfect knowledge of the equipment
and machines they use. They should be aware of quality
control essentials and be well versed with interpretational
aspects of the reports generated by their laboratories. In
addition to what has been mentioned under primary
laboratories, secondary laboratories also perform:
¾¾ Routine immunohematological tests.
¾¾ Routine examination of all body fluids, e.g. semen,
cerebrospinal fluid (CSF), sputum, etc.
¾¾ Routine bacteriologic studies including stains, cultures
and antibiograms. Routine mycological investigations

would include—primary cultures, isolation and identi­
fication techniques along with microscopic evaluation.
¾¾ Routine immunoserological tests. These can include
tests like Widal, STS, ELISA or strip or device tests HIV I
and II, hepatitis B and hepatitis C, etc.
¾¾ Routine biochemistry investigation and organ profile
tests, e.g. lipid, cardiac, liver and renal profiles.
¾¾ Under hematology, these laboratories may also
provide RBC indices, platelet, reticulocyte count and
absolute eosinophil counts. They can also classify
anemias and should be able to indicate hematologic
malignancies. When headed by a pathologist, they
should be in a position to report bone marrow smears/
preparation too.

3

Tertiary Laboratory
These kinds of laboratories should be able to perform all
kinds of sophisticated and delicate/precise investigations.
The tertiary laboratories can branch out in very special
fields and not cater to all aspects of specialized tests.
Besides doing all investigations that are conducted in
secondary laboratories, they also carry out the following:
¾¾ Specialized hematological (e.g. leukemia type), coagu­
lation profiles and immunohematological investigations.
They are equipped with 18 parameter cell counters with
differentials and flow cytometry
¾¾ Complete biochemical assays, commonly referred to
as SMA-12, SMA 27, etc. Also included are elemental

assays, e.g. zinc, magnesium, iron, total iron binding
capacity (TIBC), lithium, etc. special enzymes like
HBDH, lipase and isoenzymes, etc.
¾¾ Complete immunology based assays for hormones,
cancer markers, hepatitis markers, rheumatic/auto­
immunity etiology-based profiles, TORCH profiles,
rare infectious diseases (e.g. brucellosis leptospirosis,
cysticercosis, echinococcosis, etc.)
¾¾ All microbiological processes, e.g. cultures—aerobic,
anaerobic, fungal, tubercular, etc. with antibiograms.
The techniques for these investigations may vary. They
may be ELISA, chemiluminescence, turbidimetry, PCR,
etc. These laboratories are totally automated and have
sizable workload. Furthermore, they also undertake all
histopathology (simple H and E, special staining techniques,
immunohistochemistry methods) and cytopathology
processing and reportings. They may also undertake
cytogenetic investigations, e.g. chromosomal analysis.
The dissemination of reports from these laboratories is in
keeping with recent trends in telecommunications, e.g. fax,
e-mail, etc.
In the United States of America, these laboratories
though classified differently (with a few differences) are
covered under the Clinical Laboratory Improvement Act
(CLIA) of 1988.

LABORATORY SET-UP
Unless the laboratory is hygienic and provides necessary
physical and operative comfort, it would be wrong to
expect perfect results. To get perfect results, one has to

provide a perfect set-up for people to work in.

Laboratory Building and Space
Ample working space is absolutely essential. For smaller
laboratories up to 25 square meters (Fig. 1.2), the working
platforms can be arranged along the walls while the central
area is kept free for movement.


4

Concise Book of Medical Laboratory Technology: Methods and Interpretations
from slides or washing glassware or discharging noncontaminated laboratory refuse.

Physical Aspects of a Laboratory

FIG. 1.2: A typical small laboratory

For larger areas, partitions can be made which would
create separate spaces for different sections (Fig. 1.3).
The chief pathologist must have casual access to all subunits of the laboratory. If possible, he should be able to
directly see into the cabins either through glass windows
or through closed circuit cameras. In the cabins again, the
central region should be kept free and benches be placed
against the walls and away from the doors.
¾¾ Hygiene is of utmost importance. The whole facility
should be absolutely clean, uncrowded and devoid of
any hindrances to movement of men and materials.
Never, should a chance arise where two people would
clash or contaminated material would be spilt all over

¾¾ Scratch proof matt finish vitrified floor (slip resistant)
should be provided. The walls should preferably have
white ceramic tiles. Such provisions are resistant to
chemicals and disinfectants
¾¾ All benches should be preferably 2½ feet high and those
to be used while standing should be at least 3 feet high.
The bench surfaces should be solvent and acid proof.
Every laboratory and/or its section must have at least
one sink and one hand wash basin. The hand wash basin
should not be used for any other purpose, the sink can
be utilized for laboratory purposes like washing off stains

Sero-immunology
ELISA’s, PCRs, drugs,
Cancer markers

Biochemistry

Microbiology

Pathologist’s chamber

Collection of
specimens and
report delivery

Hematology +
Clinical pathology

Histopathology

Cytopathology

Toilet

FIG.1.3: A typical large/complete laboratory plan

¾¾ The ambient temperature should be within the comfort
zone of a human body. It should between 21 and 27°C.
If the laboratory is in a cold zone, it must have heating
provision, and conversely, if it is in a hot zone, it must
have cooling or air conditioning. The environment
control appliances like air conditioners or heaters must
not directly discharge air at the working bench zone
¾¾ A good exhaust system is a must for all laboratories.
This removes dirty air (aerosols), which may at times
be foul smelling. The sample collection zone too, must
have excellent exhaust provision
¾¾ Adequate ventilation is also essential but without
strong currents of air
¾¾ Lighting should be more than adequate and places
where very delicate or fine processes are being
conducted should have additional lighting provision.
As far as possible, do not use excessive heat producing
bulbs and lamps. The new CFLs are ideal
¾¾ Windows that are exposed to bright sunlight can be
internally fitted with reflective films or blinds
¾¾ There should be sufficient running water for the
laboratory and all must have sufficient number of sinks
and hand wash basins
¾¾ As most machines consume a lot of electricity, sufficient

power load (a little in excess) must be available to the
laboratory

Provisions and Precautions
Every working room or cabin should have adequately
spaced provision of water, electricity, gas, sinks lighting
and exhausts. All aspects, whether plumbing, electrical
systems or gas connection must pass through regular
inspections and a log book should be maintained of such
preventive exercises. Preventive maintenance should be
carried out by knowledgeable and qualified persons.

Fire Prevention
¾¾ Install appropriate fire extinguishing system and timely
testing of such a system be conducted at regular intervals
¾¾ Color code and place firefighting equipment at an easily
visible and reachable location. Check the working
capability of all such systems at regular intervals
¾¾ Provide adequate ventilation in zones where flammable
chemicals are used. Before these substances reach
combustible or explosive concentration, they should
be removed by mechanical exhausts


Laboratory
¾¾ Post “No-smoking” signs in zones where smoking can
be hazardous
¾¾ Lastly, mark clearly the emergency exit points. Keep the
emergency exit route free from obstructions.

5

Staff Safety and Facilities

¾¾ Label all bottles with proper names of contents and
affix warning signs and symbols as applicable to them
¾¾ Clearly display the warning charts (both chemical and
radioactive) next to such containers. All staff members
working in such areas should be well trained to handle
accidents of any kind that can happen
¾¾ A stringent record of stocks should be maintained of
all persons and radioactive substances being used in
the laboratory. A bottle lost or stolen is invitation to
problem.

The most important asset of any institution is the man­
power that works for it. It holds true for laboratories
too. Absence of staff due to morbidity or mortality can
stifle your working capacity, capability and reputation.
Provide adequate facilities to your team. (Designate a
room or space meant exclusively for retiring or resting and
consuming foodstuffs).
¾¾ Hot and cold running water with soap and disinfectants
should always be provided. Clean hand towels should
be replaced daily
¾¾ A clean toilet for use by staff members is mandatory as
are the changing rooms. If possible, separate units for
male and female members should be provided
¾¾ Biomedical wastes and non-biomedical wastes should
be discarded properly and safely. Chemical treatment

of liquid wastes and incineration of solid wastes should
not be overlooked. Wastes handled properly ensures
good health of your working team
¾¾ Designate a room or space meant exclusively for
retiring or resting and consuming foodstuffs. Under no
circumstances, laboratorians should eat or drink on
their workbenches. Provide safe drinking water to all
¾¾ Each room/cabin must have a first-aid box kept at an
identified place that is easily accessible. Every person
working in the laboratory must be aware of all hazards
that exist and must also know about the remedial
measures that should be taken if something happens.
What can be managed in house should be managed,
when required, assistance of other specialists must be
taken. Contact numbers of such institutions/specialists
must be displayed prominently
¾¾ All members of your team must be immunized as
relevant to the laboratory work. Make sure no single
person works alone in a room or cabin. Two compatible
persons should work together always.

Stores

Basic Laboratory Safety

Electrical Installations
¾¾ Hire a proper, qualified electrical engineer and explain
to him the purpose of the premises being taken. As far
as possible, all points where sparks can be generated
should be kept out of room/cabins where explosive

chemicals are likely to be used
¾¾ Use earthing everywhere and install fire-resistant
cables in the laboratory
¾¾ Employ only certified products
¾¾ Use one electrical socket for a single device or machine.
Overloading is usually the cause of accidents.

Liquefied and Compressed Gases
¾¾ Color code and identify each gas container. Check their
valves regularly
¾¾ Keep all such cylinders away from sources of heat and
electrical sparks
¾¾ When not in use, replace protection/safety caps back
on the cylinder mouths.

Chemicals and Radioactive Substances

¾¾ Every bottle/container should be labeled. Affix the
hazard intensity on the bottle or the container
¾¾ Ensure in every possible way that the containers cannot
under any circumstances fall or spill. This can be done
by placing the most dangerous chemical at the bottom
or at the floor level
¾¾ Proper ventilation should be ensured in storage
zones that house flammable chemicals. Keep fire
extinguishing equipment handy. Post “No smoking”
signs that are clearly visible. Make sure that the place
remains free from pests.

¾¾ Use only certified safe equipment in the laboratory

¾¾ Decontaminate all equipment regularly and before
their servicing or maintenance, use appropriate
disinfectants correctly
¾¾ As far as possible, use disposable plasticware to
avoid contamination (chemical, biological, etc.) and
breakages with ensuing dangers
¾¾ Regularly test and service biological safety cabinets and
fume cupboards.
Appropriate safety measures taken by you will go a long
way in enhancing productivity.


6

Concise Book of Medical Laboratory Technology: Methods and Interpretations

As a rule, the place for receiving or withdrawing the
specimens should be separate from the working compart­
ment. To avoid specimen mixing (hazardous), each sample
should be carefully labeled. The label should clearly
mention the alloted specimen number, the date and time
of receipt of specimen, the investigations to be done and
most important the name of the patient.
Both, the clinical and the paraclinical workers are
equally at risk of acquiring transmissible diseases through
the patient or through the test samples. The risk of these
can be lessened by taking appropriate vaccinations. In
addition, one should attend to one’s general hygiene and
prevent fomite transmission of any infectious disease.
Disinfect the working benches and as far as possible

autoclave (or chemically disinfect) various glassware used
in the laboratory. Use a rubber teat for sucking/filling
the pipettes. To avoid strain on the eyes, keep both eyes
open while doing microscopic work. Before leaving the
laboratory, one should thoroughly wash one’s hands with
soap and water, and then rinse them well in a disinfectant
lotion.

CODE OF CONDUCT FOR MEDICAL
LABORATORY PERSONNEL
1. Place the well-being and service of the sick above your
own interests.
2. Be loyal to your medical laboratory profession by
maintaining high standards of work and strive to
improve your professional knowledge.
3. Work scientifically and with complete honesty.
4. Do not misuse your professional skills or knowledge
for personal gain.
5. Never take anything from your place of work that does
not belong to you.
6. Do not disclose to a patient or any unauthorized
person the result of your investigations.
7. Treat with utmost confidentiality and personal
information that you may learn about a patient.
8. Respect and work in harmony with the other members
of your hospital staff or health center team.
9. Be at all times courteous, patient, and considerate to
the sick and their relations.
10. Promote health care and the prevention and control
of disease.

11. Follow safety procedures and know how to apply first
aid.
12. Do not drink alcohol during laboratory working hours
or when on emergency stand-by.
13. Use equipment and laboratory-ware correctly and with
care.

14. Do not waste reagents or other laboratory supplies.
15. Fulfil reliably and completely the terms and conditions
of your employment.
Always remember that you can be a patient tomorrow.
Treat others as you would want them to treat you.

ACCIDENTS
Safety Measures in the Laboratory
You must remain alert and cautious while working in
the laboratory. You must know that careless handling
of reagents, glassware or specimens to be tested in the
laboratory can cause serious injury and is dangerous to
life.

Hazards in the Clinical Laboratory
Clinical laboratory workers may encounter three types of
hazards:
1.Physical,
2. Chemical, and
3. Biological hazards.

Physical Hazards
Physical hazards are present in ordinary equipment

or surroundings. Electrical equipment, open flames,
laboratory instruments and glassware can all be hazardous
if improperly used.
Electricity
¾¾ All electrical equipment must be properly grounded
following the manufacturer’s instructions
¾¾ Even minor repairs, such as replacement of the micro­
scope bulbs, require that instrument be disconnected
from the power supply before the work is begun
¾¾ All electrical cords and plugs be kept in good shape and
order with no frayed cords or exposed wires
¾¾ Avoid overloaded circuits
¾¾ Extension cords present several safety hazards and
should not be used except in emergency.
Fire
Fire is a potential danger in the workplace:♥
¾¾ Though rare, they can occur when open flames are
used in the vicinity of flammable liquids
¾¾ Make sure that loose clothing and long hair do not
catch fire
¾¾ Instead of open flames, use hot plates, microwave
ovens, electric incinerators and slide warmers
¾¾ Store flammable chemicals in a flameproof cabinet,
away from heat sources and well-ventilated area. A
flameproof cabinet can protect flammable chemical


Laboratory
from flames until firefighters arrive and also allow
workers more time to escape

¾¾ All laboratory workers must know about the escape
route and procedure to follow if that exit is blocked
¾¾ All workers must know the location of fire extinguishers
and how to use them
¾¾ Inspect all fire extinguishers periodically and log the
date of inspection.
Usual Causes of Fire in the Laboratory
¾¾ Naked flames (do not work with loose clothing and
long hair near naked flames). Naked flames can also
ignite flammable liquids and gases
¾¾ Electrical overloading. Use one socket for one
equipment only. Do not operate a 15 amp equipment
from a 5 amp socket
¾¾ Poor electrical maintenance. No frayed or open/
exposed wires be ever used
¾¾ Leaving equipment switched when not in use. Out of
sight is out of mind
¾¾ Deteriorated gas tubing. Leakage of gas is an open
invitation to fire hazard. If you suspect gas leakage,
do not operate any electrical equipment (do not ever
switch on a light or a fan)
¾¾ Smoking in the laboratory
¾¾ Misusing matches. Use carbonized matches as far as
possible
¾¾ Storing flammable and explosive chemicals in an
ordinary refrigerator.

When a Fire Occur
¾¾ For tiny blazes; water, sand and a fire blanket can be
employed to put out the fire. For larger blaze, a fire

extinguisher can be used
¾¾ Never use water on an electrical fire or one caused
by organic solvents (ether, alcohol, petrol, etc.). For
electrical fires, use carbon dioxide fire extinguisher. For
organic solvents, use sand or halon
¾¾ Escape via the fire exit route. Stay close to the floor,
cover your mouth and nose with a damp cloth to filter
out some of the harmful fumes
¾¾ Inform firefighting department of your area if you
feel the fire can go out of hand. Medium to large fires
should be reported irrespective of your preparedness to
handle them.
Laboratory Equipment (Table 1.1)
¾¾ Use all laboratory equipment as per manufacturer’s
recommendation
¾¾ Any instrument with moving parts, such as a centrifuge,
must be operated with a special regard for safety. Latch,

7

the lid before turning it on. On turning it off, do not
open the lid before it has come to a complete stop
¾¾ Autoclaves present special hazards. Strictly adhere to
manufacturer’s instruction to prevent explosions and
burns. Use insulated gloves while removing hot items
from the autoclave.
Glassware
¾¾ Use glassware that is free of chips and cracks. Damaged
glassware is weakened and may break, resulting in
injury

¾¾ Broken glass should be cleaned with a brush and
dustpan and not with bare hands
¾¾ Glass should not be discarded into regular trashcans,
but into rigid cardboard or plastic containers
¾¾ Wherever possible, replace glassware with plasticware.
Equipment Related Hazards
¾¾ Hypodermic needles: Accidental inoculation, aerosol
or spillage
¾¾ Centrifuges: Aerosols, splashing and tube breakages
¾¾ Culture stirrers, shakers, agitators: Aerosols, splashing
and spillage
¾¾ Refrigeration: If flammable chemicals are stored within
them, the light switches, thermostats, etc. can provide
sparks to ignite them
¾¾ Water baths: Provide ground for microorganismal
growth
(The risk of acquiring hepatitis B from a needle stick is
30%, hepatitis C is 2 to 10% and HIV is 0.3%).
Equipment/Materials Employed to Eliminate/Reduce
Hazards
¾¾ Laboratory apron: Assists in diminishing skin contacts
to a certain extent
TABLE 1.1: Fire fighting equipment

Fire fighting material Used for

Contraindicated for

Fire blanket

Clothing fire,
G small blaze

Electrical fires, flammable liquids, a small
blaze burning metals,
alkali metal

Water

Paper, wood, fabric

Electrical fires, flammable liquids, burning metal, alkali metal

CO2 fire
extinguisher

Flammable liquids —
and gases, electrical fire

Dry powder

As above

—

Foam

Flammable liquids

—

Halon spray

All kinds of fires

—


8

Concise Book of Medical Laboratory Technology: Methods and Interpretations

¾¾ Biological safety cabinets: Prevent dangers arising out
of aerosols and splatters
¾¾ Splatter shields: Provide protection from splatter of
specimen and chemicals
¾¾ Pipetting aids (teat or electromechanical devices).
Prevent from hazards arising out of mouth pipetting
¾¾ Goggles: Protect eyes from impacts and splashes
¾¾ Face shields: Protect the face from impacts and
splashes.

¾¾ Chlorine—with ammonia, hydrogen, benzene and
other finely divided metals
¾¾ Copper—with azides, hydrogen peroxide and acetylene
¾¾ Cyanides—with all acids and alkalies
¾¾ Hydrogen peroxide—with copper, iron, chromium and
most other metals
¾¾ Iodine—with acetylene and ammonia
¾¾ Sodium azide—with lead, copper and other metals.

Safety with Chemicals/Reagents

These include ether, xylene, toluene, methanol, ethanol,
glacial acetic acid, acetic acid, acetone, acetic anhydride,
alcoholic Romanowsky stains and acid alcohol, etc.

Excepting just a couple of reagents, almost all chemicals/
reagents used even in the most basic laboratory are lethal
poisons if consumed by anyone. Even if they are splashed
on the skin/eye, they can cause irreversible damage. There
is an appropriate way of handling and storage of hazardous
chemicals to avoid injury and damage to self and others.
In our country (and other tropical nations), excessive heat
can decompose many chemicals, cause explosions, or lead
to the formation of toxic fumes.

Labeling of Hazardous Reagents/Chemicals
At appropriate places, display the prohibition signs; and
on all dangerous reagents or chemicals, stick Hazard
warning symbols. In the following pages, important signs
and symbols as related to safety in the laboratory are given.

Incompatible Chemicals
Fair number of common laboratory chemicals react
dangerously if they come in contact with specific chemicals.
Ensure that you keep such chemicals away from each other.
A few examples are listed below:
Acids
¾¾ Acetic acid with chromic acid, nitric acid, hydroxyl

compounds, ethylene glycol, peroxides and permanganates
¾¾ Chromic acid—with acetic acid, alcohol, glycerol and
other flammable liquids
¾¾ Sulfuric acid—with chlorates, perchlorates, permanganates and water.
Vaporizing Substances
¾¾ Acetone—with sulfuric acid and nitric acid
¾¾ Flammable liquids—with chromic acid, hydrogen
peroxide, nitric acid, ammonium nitrate and halogens.
Others
¾¾ Alkali metals, e.g. calcium, potassium, sodium (these
form hydroxides on coming in contact with water) and
with other chlorinated hydrocarbons

Flammable Chemicals

Storage
These should be stored in a fire-proof metal box at ground
level, preferably in a cool store. A container well lined with
tin foil can also be used. Store only small quantities of such
solvents on the shelves.
Safe Use
Ensure that there is no open flame nearby while opening a
bottle containing flammable solvent. Nearest flame should
be at least 10 feet away. Never heat a flammable liquid over
any flame. Use a water bath or electric hot plate.
Control of Fire Caused by Flammable Chemicals
Best controlled by smothering them. Use sand, thick blanket
or the now available multipurpose fire extinguishers.
Pouring water on such fires will spread them. Every
laboratory should be equipped with the commercially

available fire extinguishers. If these are not available, there
should be sand buckets in accessible places.

Corrosive Chemicals
These include strong acids, e.g. concentrated sulfuric
acid, hydrochloric acid, nitric acid, glacial acetic acid,
trichloroacetic acid, orthophosphoric acid, and strong
alkalies like sodium hydroxide and potassium hydroxide.
Storage
Store these at low levels.
Safe Use
Never attempt mouth pipetting. Accidental swallowing
can be lethal as these chemicals cause destruction of
living tissue. Always pour a corrosive chemical at below
eye level, slowly, and with great care to avoid splashing.
Wear protective eye glasses/eye shields while opening
such containers. Always add the corrosive substance to
water and that too slowly. The addition of small amount of
water to sulfuric acid is enough to produce sufficient heat
to break a glass container.


Laboratory
Toxic, Harmful, and Irritating Chemicals
These are chemicals that can cause death or serious illhealth if swallowed or inhaled or if they come in contact
with skin. Examples are potassium cyanide, mercuric
nitrate, sodium azide, sodium nitroprusside, formaldehyde
solution, chloroform, barium chloride and methanol.
Iodine and sulfuric acid also fall in this category. Skin and
mucous membrane irritants are xylene, formaldehyde and

ammonia vapors.

Storage
Store highly toxic chemicals, e.g. potassium cyanide in a
locked cupboard. Stock solutions should also be stored
safely in a cupboard, not on an open shelf.

9

Safe Use
Always wear protective gloves and after working with
them immediately lock them up. Always wash your
hands after using a toxic or harmful chemical. Keep fume
forming chemicals in a fume cupboard. Never mouth
pipette them.

Oxidizing Chemicals
These include chlorates, perchlorates, strong peroxides,
potassium dichromate, and chromic acid.
Storage
Keep these away from organic materials and reducing
agents. They can produce much heat when in contact with
other chemicals, especially flammable chemicals.

SIGNS FOR MEDICAL LABORATORIES

FIG. : General laboratory