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Clinically Pertinent Cytological Diff-Quick and Gram Stain
Clinically Pertinent Cytological Diff-Quick and Gram Stain Evaluation for the Reptilian Practitioner Kendal E Harr, DVM, MS, Dipl ACVP (Clinical Pathology), April Romagnano, PhD, DVM, DABVP (Avian) Session #214 Affiliation: URIKA, LLC, Mukilteo, WA 98275, USA (Harr), Avian and Exotic Clinic of Palm City, Palm City, Florida and the Animal Health Clinic, Jupiter, FL 33458, USA (Romagnano). Abstract: The goals of this work were to: 1) improve knowledge of preanalytic sampling techniques including blood smears, fine needle aspirates, imprints, smears, and fluid preparation including oral, dermal and cloacal swabs, cystic and solid mass sampling, joint fluids and effusions, and fecal smears and floats; and 2) enable the reptilian practitioner to better identify basic cells, classify disease processes, as well as infectious agents such as bacteria, fungi, and other structures. Discussion of diagnoses and treatment will follow. Generalized disease processes cross species and classes. The most important rule for cytologic interpretation is to not overinterpret the cytologic findings. Cytology helps guide therapeutic decision making by classification of disease process as neoplasia, fungal infection, etc but may not provide a definitive diagnosis. One should only interpret to the correct level of diagnosis and know when to refer the cytology and biopsy the lesion. Preanalytical Blood collection Collect less than < 1% of a reptile’s body weight. Use heparinized, size appropriate pediatric microtainers or Capijects®. Use the jugular vein in species where possible as the large bore vein decreases the likelihood of lymph dilution common in samples from the caudal vein. The right jugular may be larger in some species of lizard and tortoise but the size difference is not as dramatic as in avian species. -
Wright's Stain
WRIGHT’S STAIN - For in vitro use only - Catalogue No. SW80 Our Wright’s Stain can be used to stain blood Interpretation of Results smears in the detection of blood parasites. Wright’s Stain is named for James Homer If malaria parasites are present, the Wright, who devised the stain in 1902 based on a cytoplasm stains pale blue and the nuclear modification of the Romanowsky stain. The stain material stains red. Schüffner’s dots and other distinguishes easily between blood cells and RBC inclusions usually do not stain or stain became widely used for performing differential very pale with Wright’s stain. Nuclear and white blood cell counts, which are routinely cytoplasmic colors that are seen in the malarial ordered when infections are expected. The stain parasites will also be seen in the trypanosomes contains a fixative, methanol, and the stain in one and any intracellular leishmaniae that are solution. Thin films of blood are fixed with present. methanol to preserve the red cell morphology so Refer to an appropriate text for a detailed that the relationship between parasites to the red description of characteristic morphological cells can be seen clearly. structures of different parasitic organisms and human cell types. Formula per Litre • Make sure all slides are clean prior to Wright’s Stain .............................................. 1.8 g making the blood smear to ensure that the Methanol .................................................. 1000 mL stain absorbs properly • Tap water is unacceptable for the rinsing Recommended Procedure solution as the chlorine may bleach the stain 1. Dip slide for a few seconds in methanol as a fixative step and allow slide to air dry • Finding no parasites in one set of blood completely. -
Appendix A: Standardization of Staining Methods
Appendix A: Standardization of Staining Methods H. Lyon, D. Wittekind, E. Schulte No detailed descriptions of staining methods are provided in this book. The reader is referred to one or more of the excellent texts covering this field (cf. Preface). Nevertheless, we have feIt it appropriate to inc1ude this appendix which gives some of our views on the technical aspects of procedures which should be given particular emphasis. It is our opinion that the need for standardization and quan titative methods in daily work is pressing. This appendix sets out the appropriate general considerations followed by a few selected methods in order to cover this area. A.l General Considerations According to Boon and Wittekind (1986) the principle aim of standardizing staining methods is to render their application reproducible and therefore reliable. This is of the utmost importance when dyes and stains are used for automated cell pattern recognition (Wittekind, 1985; Wittekind and Schulte, 1987). The theoretical background for standardization of cell and tissue preparation sterns from the fact that any preparatory step - from cell sampling to mounting of the stained slide - will somehow affect the structure of the cell and ultimately lead to the production of a particular staining pattern which, in strict tenns, is an artifact. What we eventually observe by microscopy is, from the perspective of a cell, the product of a rather violent procedure: In cytological preparations the cells have been isolated from their tissue, spread out on the surface of a glass slide and immersed in a liquid poison which abruptly arrests and - sensu stricto - "fixes" the cell in the very last moment of its life. -
Special Techniques Applicable to Bone Marrow Diagnosis
TWO SPECIAL TECHNIQUES APPLICABLE TO BONE MARROW DIAGNOSIS Peripheral blood samples, bone marrow aspirates and niques that may be applied to trephine biopsy trephine biopsy specimens are suitable for many sections include: (i) a wider range of cytochemical diagnostic investigations, in addition to routine stains; (ii) immunohistochemistry; (iii) cytogenetic microscopy of Romanowsky-stained blood and bone and molecular genetic analysis; and (iv) ultrastruc- marrow films and haematoxylin and eosin-stained tural examination. histological sections. Some of these techniques, for example Perls’ stain to demonstrate haemosiderin Cytochemical and histochemical stains in a bone marrow aspirate, are so often useful that they are performed routinely, whereas other tech- Cytochemical stains on bone marrow niques are applied selectively. This chapter will deal aspirates predominantly with special techniques that are applicable to bone marrow aspirates and trephine Perls’ stain for iron biopsy sections but reference will be made to the peripheral blood where this is the more appropriate A Perls’ or Prussian blue stain (Figs 2.1 and 2.2) tissue for study. demonstrates haemosiderin in bone marrow macro- Bone marrow aspirate films are stained routinely phages and within erythroblasts. Consequently, it with a Romanowsky stain such as a May– allows assessment of both the amount of iron in Grünwald–Giemsa (MGG) or a Wright–Giemsa stain. reticulo-endothelial stores and the availability of Other diagnostic procedures that may be of use iron to developing erythroblasts. in individual cases include: (i) cytochemistry; (ii) Assessment of storage iron requires that an ade- immunophenotyping (by immunocytochemistry or quate number of fragments are obtained. A bone flow cytometry); (iii) cytogenetic and molecular marrow film or squash will contain both intracellu- genetic analysis; (iv) ultrastructural examination; lar and extracellular iron, the latter being derived (v) culture for micro-organisms; and (vi) culture for from crushed macrophages. -
Blood Film Preparation and Staining Procedures
INTERPRETATION OF THE PERIPHERAL 00. ם BLOOD FILM 0272–2712/02 $15.00 BLOOD FILM PREPARATION AND STAINING PROCEDURES Berend Houwen, MD, PhD The blood film is one of the world’s most widely and frequently used tests and has undergone remarkably few changes since its introduction as a clinical diagnostic tool in the late 1800s. The origins of stained blood film microscopy are somewhat obscure, and a clear reference for the ‘‘first person or persons’’ to describe its use as a clinical laboratory procedure cannot be established with certainty. Antonie van Leeuwenhoek, in the seventeenth century, was the first to describe blood cells, using whole blood preparations and not blood films for his observations on blood corpuscles. In fact, he would not have been able to use blood film–based microscopy because it requires considerably more sophisti- cated optics than were available in his day. A second technologic innovation enabling blood film microscopy was the introduction of aniline dyes in the second half of the nineteenth century. This made it possible to study individual blood cells by light microscopy after a small amount of blood had been placed and smeared onto glass slides, dried, fixed, and then stained. Paul Ehrlich introduced eosin as the first of these dyes for staining blood films in 1856, followed by hematoxylin in 1865, and later by the metachromatic Romanowsky dyes. A few refinements have since been made, consisting mainly of improve- ments in dye quality, staining procedures, automation of slide preparation, and staining, but the basic elements of blood film preparation and analysis have not changed for over a century. -
Dr.Sithy Athiya Munavarah Dr. Johnsy Merla J* Original Research Paper
Original Research Paper Volume-9 | Issue-2 | February-2019 | PRINT ISSN - 2249-555X Pathology CYTOMORPHOLOGY OF NODULAR THYROID LESIONS : A COMPARATIVE ANALYSIS OF WET AND AIR DRIED SMEARS Dr.Sithy Athiya PG, Director & HOD, Department of pathology,Karpaga Vinayaga Institute of Medical Munavarah Sciences&Reseacrh center Dr. Meenakshi Assistant Professor , Department of pathology, Karur Medical College. Dr. Suresh Durai J Professor, Department of pathology, Tirunelveli Medical College Dr. Johnsy Merla Assistant Professor, Department of pathology, Tirunelveli Medical College J* *Corresponding Author Dr. Chandru Mari Assistant Professor, Department of pathology, Tirunelveli Medical College Dr. Shantaraman Professor&HOD, Department of pathology, Tirunelveli Medical College. K ABSTRACT FNAC of thyroid lesions have sensitivity as high as 93.4% with a positive predictive value of malignancy 98.6 % and 74.9 %specicity. Two fundamentally different methods of xation and staining are used in FNAC: air-drying followed by a Romanowsky stain such as May Grunwalds Gimsa (MGG), Jenner-Giemsa, Wright's stain or Diff-Quik; and alcohol-xation followed by Papanicolaou (Pap) or hematoxylin and eosin (H&E) staining. Combining the morphological features of various stains often improve the diagnostic accuracy.In the present study, cytoplasmic granularity, paravacuolar granules and thin colloid are very well demonstrated using Wright Giemsa stain. Cell borders and crisp nuclear features such as chromatin pattern, intranuclear inclusions are best appreciated using wet xed smears stained with H&E and Pap stains. The cytomorphologic features of nodular thyroid lesions using multiple cytological staining techniques to enhance diagnostic sensitivity is evaluated in this study. KEYWORDS : : Cytology ,Fine Needle Aspiration, Romanowsky stain, Thyroid. -
Cytology Product Range
Cytology Product Range Improving Laboratory Efficiencies and Delivering Better Patient Care ProductsBlock Trimmers to Achieve Best 4-5Results CellCeps+ Heated Forceps 6-7 Wax Dispensers 8-9 Tissue Section Baths 10-11 CONTENTS ROUTINE SPECIMEN PREPARATION Slide Drying Hotplates 12-13 Cytology Funnels 3-4 CellPath’s innovative range of Cytology products are Section Dryers 14-15 Funnel Clips 5 designed to improve laboratory practices whilst ultimately Circle Slides 5 delivering better patient care. Being high quality assured, our Tissue Section Baths 10-11 LARGE VOLUME SPECIMEN PREPARATION consumables deliver improved laboratory efficiencies, better Slide Drying Hotplates 12-13 EcoFunnel Mega Disposable Cytology Funnel 6 cell collection and significantly reduced patient discomfort. EcoFunnel Mega Slide+ 6 Section Dryers 14-15 LIQUID BASED CYTOLOGY The Cytology range includes specialised collection devices CerviBrushes, Cervex Sampling Brushes to suit your purposes; funnels that are compatible with the & Aylesbury Spatula 7-9 most commonly used centrifuges, circle slides, fixatives, stains and slide storage solutions. CYTOLOGICAL FIXATIVES CytoFixx & CytoFixx II 10 UriCyte+ 11 CYTOLOGY STAINS Premium Papanicolaou Stains 12 Nuclear Stains 13 Rapid Giemsa/ May Grunwald 13 ‘Three Step’ Staining Kits VFM Papanicolaou Stains 13 SLIDE STORAGE FiloSlide Slide Trays 14 FiloSlide Wallets 15 FiloSlide Cabinet 16 Based in the UK, CellPath Ltd is a manufacturer and distributor of pathology consumables, instruments & services. Certified to: ISO 9001 ISO 13485 2 NON-GYNAE CENTRIFUGE CONSUMABLES Our non-gynaecological consumables provide solutions for both routine and large volume sample preparation. Funnels are fully compatible with the Shandon CytoSpin® and related circle slides are also specifically for use with a centrifuge. -
Diagnostic Stains & Laboratory Reagents
polysciences.com Diagnostic Stains & Laboratory Reagents • Cytology • Dermatology • General Reagents • Hematology • Histology • Microbiology U.S. Corporate Headquarters | 400 Valley Rd, Warrington, PA 18976 | 1(800) 523-2575 (215) 343-6484 | Fax 1(800) 343-3291 | [email protected] Polysciences Europe GmbH | Badener Str. 13, 69493 Hirschberg an der Bergstr., Germany | +(49) 6201 845 20 0 | Fax +(49) 6201 845 20 20 | [email protected] Polysciences Asia Pacific, Inc. | 2F-1, 207 DunHua N. Rd. Taipei, Taiwan 10595 | (886) 2 8712 0600 | Fax (886) 2 8712 2677 | [email protected] Life Sciences Astral Diagnostics Catalog # Size Cytology Acrylic Mounting Medium . 25835-16 16 oz Histo/Cyto Mounting Medium in a toluene matrix using an acrylic resin for coverslipping . 25835-4 4 oz AQUAbluing Reagent . 25836-1 1 gal Bluing reagent used to provide crisp nuclear detail . Bluing Reagent . 25837-32 32 oz Lithium carbonate solution used to provide crisp nucler detail to cells . 25837-1 1 gal Human Colon, 20X Harris Hematoxylin, Eosin Y Pap Stain Gynecological stain used in combination with OG-6 and hematoxylin in the diagnosis of malignant cytological diseases . FDA approved for in vitro diagnostic use . EA-36 25865-32 32 oz 25865-1 1 gal EA-50 25841-2 .5 2 .5 gal EA-65 25842-2 .5 2 5. gal EASYpap . 25843-32 32 oz Gynecological single solution counterstain . A substitute to the traditional Eosin 25843-1 1 gal azure and Orange G . FDA approved for in vitro diagnostic use . Formalin 10%, Buffered . 25848-16 16 oz Most widely used fixative used for routine processing of tissue in histology laboratories . -
Blood Products
Blood Products Horse Blood Code Description Size Price £ HB033 Defibrinated 25ml 3.85 HB034 Defibrinated 100ml 8.85 HB030 Defibrinated 200ml 15.90 HB029 Defibrinated 300ml 21.80 HB032 Defibrinated 400ml 29.50 HB035 Defibrinated 500ml 36.70 HB001 Defibrinated 1 Litre (bag) 60.55 HB036 *Lysed 25ml 5.00 HB037 *Lysed 100ml 11.20 HB025 *Lysed 200ml 20.45 HB026 *Lysed 300ml 28.85 HB024 *Lysed 400ml 36.25 HB038 *Lysed 500ml 50.70 HB056 *Lysed 1 Litre (bag) 83.75 HB039 Oxalated 25ml 4.25 HB040 Oxalated 100ml 9.05 HB041 Oxalated 500ml 39.60 HB042 Formolised 25ml 4.25 HB043 Formolised 100ml 9.00 HB044 Formolised 500ml 39.25 HB045 ACD 25ml 4.25 HB046 ACD 100ml 9.05 HB047 ACD 500ml 39.25 HB048 Alsever's 25ml 2.90 HB049 Alsever's 100ml 7.50 HB050 Alsever's 500ml 32.30 *Products are lysed with the addition of saponin. Other anticoagulants available, please ask. Sheep Blood Code Description Size Price £ SB053 Defibrinated 25ml 5.55 SB054 Defibrinated 100ml 14.60 SB055 Defibrinated 500ml 61.75 SB056 *Lysed 25ml POA SB057 *Lysed 100ml POA SB058 *Lysed 500ml POA SB059 Oxalated 25ml POA SB060 Oxalated 100ml POA SB061 Oxalated 500ml POA SB062 Formolised 25ml POA SB063 Formolised 100ml POA SB064 Formolised 500ml POA SB065 ACD 25ml POA SB066 ACD 100ml POA SB067 ACD 500ml POA SB068 Alsever's 25ml 4.25 SB069 Alsever's 100ml 10.35 SB070 Alsever's 500ml 45.55 *Products are lysed with the addition of saponin. Other anticoagulants available, please ask. Blood Products continued Chicken Blood Cells (Adult) Code Description Size Price £ FB010 Alsever's 10ml 34.80 FB011 Alsever's 25ml 57.55 FB012 Alsever's 50ml 93.85 FB013 Alsever's 100ml 154.40 Adult Chicken Blood Cells in Alsever’s is a Cell product. -
Assessment of Different Stains and Staining Procedures
ASSESSMENT OF DIFFERENT STAINS AND STAINING PROCEDURES FOR MICROSCOPIC DETECTION OF MALARIA PARASITES By: Awad Alla Hamza Osman Kashif BSc of Medical Laboratory Sciences (1995) A thesis submitted for fulfillment requirement of master degree in Medical Laboratory Sciences (Medical Parasitology) At the Department of Parasitology & Medical Entomology Faculty of Medical Laboratory Sciences University of Khartoum Supervisor: Dr. Eldirdieri Salim Ahmed College of Medicine –University of Juba September 2004 LIST OF CONTENTS DEDICATION………………………………………………………………….. I ACKNOWLEDGEMENT.…………………………………………………….. II LIST OF TABLES……………………………………………………………... III LIST OF FIGURES……………………………………………………………. IV LIST OF APPENDICES……………………………………………………….. V ABSTRACT…………………………………………………………………….. VI ABSTRACT (ARABIC)………………………………………………………... VII 1- INTRODUCTION & LITREATURE REVIEW 1 1.1 General introduction……………………………………………………….…... 1 1.2 Historical background…………………………………………………………. 1 1.3 Malaria parasites………………………………………………………………. 3 1.4 Transmission of malaria……………………………………………………….. 4 1.5 The life cycle of malaria……………………………………………………….. 8 1.5.1 Pre-erythrocytic phase……………………………………………………….. 8 1.5.2 Erythrocytic phase…………………………………………………………… 9 1.5.3 Vector phase (Sporogony)……………………………………………….…... 9 1.6 Pathogenesis of malaria………………………………………………………... 12 1.7 Immunity to malaria…………………………………………………………… 14 1.8 Diagnosis of malaria……………………………………………………….…... 15 1.8.1 Clinical diagnosis……………………………………………………………. 15 1.8.2 Microscopic diagnosis……………………………………………………….. 16 1.8.2.1 Conventional thick and thin blood smears………………………………… 18 1.8.2.1.1 Blood drawing technique…………………………………………….….. 19 1.8.2.1.2 Preparation of the smears…………………………………………….….. 19 1.8.2.1.3 Staining procedures…………………………………………………. 21 1.8.2.1.4 Microscopic examination. ………………………………………….. 23 1.8.2.2 Quantitative Buffy Coat (QBC) Test…………………………………. 24 1.8.2. 3 Saponin lyzing technique…………………………………………….. 24 1.8.3 Rapid Diagnostic Tests (RDTs)………………………………………… 25 1.8.4 Deoxyribonucleic acid (DNA) probes and Polymerase Chain Reaction 26 (PCR). -
Difference Between Giemsa Stain and Wright Stain
Difference Between Giemsa Stain and Wright Stain www.differencebetween.com Key Difference – Giemsa Stain vs Wright Stain In the context of microscopy, staining is considered as an essential step during enhancement of the contrast of the microscopic image, especially to highlight different structures in biological tissues. During the staining of peripheral blood and bone marrow smears, Wright and Giemsa stains are used. These stains are known as Romanowsky stains. Both these stains are composed of important components: oxidized methylene blue, eosin Y, and azure B dyes. The function of methylene blue and azure B is to stain the nucleus with colors varying from blue to purple. These stains are widely used during the study of red blood cell morphology and during the performance of differential white blood cell counts. Diagnosis of different disease conditions such as leukemia could be achieved through Romanowsky staining procedures. Wright staining is used to differentiate blood cells which consist of a mixture of eosin and methylene blue dyes. Giemsa staining is utilized during the staining of bacterial cells as well as human cells and could be combined with Wright stain to develop Giemsa Wright stain. This is the key difference between Giemsa stain and Wright stain. What is Giemsa Stain? Giemsa stain is used for cytogenetics and histopathological diagnosis of parasites of malaria and other parasitic diseases. Giemsa stain can also be considered as a basic stain in classifying lymphomas in the classification of Kiel. Giemsa stain is needed for Giemsa banding which is commonly known as G-banding. Giemsa banding is used to stain chromosomes and also used in creating karyograms. -
Product Specification Sheet
PRODUCT SPECIFICATION SHEET GIEMSA STAIN(SI015) Use Giemsa stain is a mixture of methylene blue, eosin, and azure B. It is specific for the phosphate groups of DNA and attaches itself to where there are high amounts of adenine-thymine bonding. Giemsa stain is a modified Romanowsky stain, very similar to Wright’s stain and gives the same colour reaction with the cellular components of the blood. Giemsa stain is mainly used for staining of peripheral blood smears and specimens obtained from the bone marrow. It is used to obtain differential white blood cell counts. Giemsa stain is also used in cytogenetics to stain the chromosomes and identify chromosomal aberrations. It is commonly used for G-Banding (Geimsa-Banding). Principle The polychromic staining solutions (Giemsa, Wrights) contain methylene blue and eosin. These basic and acidic dyes induce multiple colours when applied to cells. Methanol acts as fixative and also as a solvent. The fixative does not allow any further change in the cells and makes them adhere to the glass slide. The basic component of white cells (i.e. cytoplasm) is stained by acidic dye and they are described as eosinophilic or acidophilc. The acidic components (e.g. nucleus with nucleic acid) take blue to purple shades by the basic dye and they are called basophilic. The neutral components of the cell are stained by both the dyes. Formula Ingredients Formula / Litre Giemsa Stain 3.8 g Methyl Alcohol 250.0 ml Glycerol 250.0ml Precautions 1. For Invitro Diagnostic use only. 2. Observe all standard safety precautions consistent with hazard(s) stated 3.