Gst Gram Staining Learning Objectives the Student Will Use Aseptic Techniques in the Safe Inoculation of Various Forms of Media
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The Suitability of Certain Stains 3R Studying Lignification in Balsam Fir, Ibies Balsamea (L.) Mill
The suitability of certain stains 3r studying lignification in balsam fir, ibies balsamea (L.) Mill Kutscha and Gray Technical bulletin 53 March 1972 Life Sciences and Agriculture Experiment Station Cover photo: Safranin and aniline blue, showing blue unlignified cambium (top of photo) and contrasting red lignified tissue (lower part of photo). Imma ture, secondary walls appear blue-blue green and can be seen approximately five to eight cells down from the cambial region. Section of FAA-killed and celloidm embedded compression wood sample collected July 6, 1966; X 320. ABSTRACT An investigation was conducted to examine the suitability of ten staining reactions for studying lignification in balsam fir, Abies balsamea (L.) Mill. Two experiments were carried out on material collected on two different dates. In each experiment slides of fresh, FAA-killed and FAA-killed celloldin-embedded material of normal and compression wood were stained and evaluated. No significant difference in staining reactions was found between material collected on different dates. In each experiment, the embedded material showed somewhat superior results compared with the fresh and FAA-killed material with at least half of the stains. No marked difference was observed between normal and compression wood. This study emphasized the need for considsring each of the ten staining reactions on an individual basis, since each has particular ad vantages and disadvantages as emphasized throughout the study. Stain ing schedules were prepared and tables compiled to determine -
Revisions Inserts Rev from Rev to JOB
BALTSO0191 Version 11.0 Template 4 Revisions Inserts Rev from Rev to JOB # 06 07 52-17 Notes: 1. BD Catalog Number: 212525, 212526, 212527, 212528, 212531, 212532, 212539, 212542, 212543, 212544, 212545 2. Blank (Sheet) Size: Length: 25.5” Width: 22” 3. Number of Pages: 28 Number of Sheets: 1 4. Page Size: Length: 8.5” Width: 5.5” Final Folded Size: 4.25” x 5.5” 5. Ink Colors: No. of Colors: 2 PMS#: 032 Red; Standard Black 6. Printed two sides: Yes X No 7. Style (see illustrations below): # 5 W W W W W W W 8. Vendor Printed X Online/In House Printed Web 9. See specication control no. N/A for material information. 10. Graphics are approved by Becton, Dickinson and Company. Supplier has the responsibility for using the most current approved revision level. Label Design COMPANY CONFIDENTIAL. THIS DOCUMENT IS THE PROPERTY OF BECTON, DICKINSON AND Becton, Dickinson and Company Proofer COMPANY AND IS NOT TO BE USED OUTSIDE THE COMPANY WITHOUT WRITTEN PERMISSION. 7 Loveton Circle Sparks, MD 21152 USA Checked By Category and Description Sheet: 1 of 29 Part Number: Package Insert, 8820191JAA Gram Stain Kits and Reagents Scale: N/A A B Gram Stain Kits and Reagents English: pages 1 – 5 Italiano: pagine 14 – 18 8820191JAA(07) Français : pages 5 – 9 Español: páginas 19 – 23 2017-09 Deutsch: Seiten 10 – 14 Contact your local BD representative for instructions. / Свържете се с местния представител на BD за инструкзии. / Pokyny vám poskytne místní zástupce společnosti BD. / Kontakt den lokale BD repræsentant for at få instruktioner. -
Laboratory Exercises in Microbiology: Discovering the Unseen World Through Hands-On Investigation
City University of New York (CUNY) CUNY Academic Works Open Educational Resources Queensborough Community College 2016 Laboratory Exercises in Microbiology: Discovering the Unseen World Through Hands-On Investigation Joan Petersen CUNY Queensborough Community College Susan McLaughlin CUNY Queensborough Community College How does access to this work benefit ou?y Let us know! More information about this work at: https://academicworks.cuny.edu/qb_oers/16 Discover additional works at: https://academicworks.cuny.edu This work is made publicly available by the City University of New York (CUNY). Contact: [email protected] Laboratory Exercises in Microbiology: Discovering the Unseen World through Hands-On Investigation By Dr. Susan McLaughlin & Dr. Joan Petersen Queensborough Community College Laboratory Exercises in Microbiology: Discovering the Unseen World through Hands-On Investigation Table of Contents Preface………………………………………………………………………………………i Acknowledgments…………………………………………………………………………..ii Microbiology Lab Safety Instructions…………………………………………………...... iii Lab 1. Introduction to Microscopy and Diversity of Cell Types……………………......... 1 Lab 2. Introduction to Aseptic Techniques and Growth Media………………………...... 19 Lab 3. Preparation of Bacterial Smears and Introduction to Staining…………………...... 37 Lab 4. Acid fast and Endospore Staining……………………………………………......... 49 Lab 5. Metabolic Activities of Bacteria…………………………………………….…....... 59 Lab 6. Dichotomous Keys……………………………………………………………......... 77 Lab 7. The Effect of Physical Factors on Microbial Growth……………………………... 85 Lab 8. Chemical Control of Microbial Growth—Disinfectants and Antibiotics…………. 99 Lab 9. The Microbiology of Milk and Food………………………………………………. 111 Lab 10. The Eukaryotes………………………………………………………………........ 123 Lab 11. Clinical Microbiology I; Anaerobic pathogens; Vectors of Infectious Disease….. 141 Lab 12. Clinical Microbiology II—Immunology and the Biolog System………………… 153 Lab 13. Putting it all Together: Case Studies in Microbiology…………………………… 163 Appendix I. -
Francisella Tularensis 6/06 Tularemia Is a Commonly Acquired Laboratory Colony Morphology Infection; All Work on Suspect F
Francisella tularensis 6/06 Tularemia is a commonly acquired laboratory Colony Morphology infection; all work on suspect F. tularensis cultures .Aerobic, fastidious, requires cysteine for growth should be performed at minimum under BSL2 .Grows poorly on Blood Agar (BA) conditions with BSL3 practices. .Chocolate Agar (CA): tiny, grey-white, opaque A colonies, 1-2 mm ≥48hr B .Cysteine Heart Agar (CHA): greenish-blue colonies, 2-4 mm ≥48h .Colonies are butyrous and smooth Gram Stain .Tiny, 0.2–0.7 μm pleomorphic, poorly stained gram-negative coccobacilli .Mostly single cells Growth on BA (A) 48 h, (B) 72 h Biochemical/Test Reactions .Oxidase: Negative A B .Catalase: Weak positive .Urease: Negative Additional Information .Can be misidentified as: Haemophilus influenzae, Actinobacillus spp. by automated ID systems .Infective Dose: 10 colony forming units Biosafety Level 3 agent (once Francisella tularensis is . Growth on CA (A) 48 h, (B) 72 h suspected, work should only be done in a certified Class II Biosafety Cabinet) .Transmission: Inhalation, insect bite, contact with tissues or bodily fluids of infected animals .Contagious: No Acceptable Specimen Types .Tissue biopsy .Whole blood: 5-10 ml blood in EDTA, and/or Inoculated blood culture bottle Swab of lesion in transport media . Gram stain Sentinel Laboratory Rule-Out of Francisella tularensis Oxidase Little to no growth on BA >48 h Small, grey-white opaque colonies on CA after ≥48 h at 35/37ºC Positive Weak Negative Positive Catalase Tiny, pleomorphic, faintly stained, gram-negative coccobacilli (red, round, and random) Perform all additional work in a certified Class II Positive Biosafety Cabinet Weak Negative Positive *Oxidase: Negative Urease *Catalase: Weak positive *Urease: Negative *Oxidase, Catalase, and Urease: Appearances of test results are not agent-specific. -
Biofire Blood Culture Identification System (BCID) Fact Sheet
BioFire Blood Culture Identification System (BCID) Fact Sheet What is BioFire BioFire BCID is a multiplex polymerase chain reaction (PCR) test designed to BCID? identify 24 different microorganism targets and three antibiotic resistance genes from positive blood culture bottles. What is the purpose The purpose of BCID is to rapidly identify common microorganisms and of BCID? antibiotic resistance genes from positive blood cultures so that antimicrobial therapy can be quickly optimized by the physician and the antibiotic stewardship pharmacist. It is anticipated that this will result in improved patient outcomes, decreased length of stay, improved antibiotic stewardship, and decreased costs. When will BCID be BCID is performed on all initially positive blood cultures after the gram stain is routinely performed and reported. performed? When will BCID not For blood cultures on the same patient that subsequently become positive with be routinely a microorganism showing the same morphology as the initial positive blood performed? culture, BCID will not be performed. BCID will not be performed on positive blood cultures with gram positive bacilli unless Listeria is suspected. BCID will not be performed on blood culture bottles > 8 hours after becoming positive. BCID will not be performed between 10PM-7AM on weekdays and 2PM-7AM on weekends. BCID will not be performed for clinics that have specifically opted out of testing. How soon will BCID After the blood culture becomes positive and the gram stain is performed and results be available? reported, the bottle will be sent to the core Microbiology lab by routine courier. BCID testing will then be performed. It is anticipated that total turnaround time will generally be 2-3 hours after the gram stain is reported. -
The Use of Matrix-Assisted Laser Desorption Ionization
Session: OS202 MALDI-TOF - diagnostics for the micro lab in the 21st century Category: 4b. Diagnostic bacteriology – non-culture based, including molecular and MALDI-TOF 25 April 2017, 14:30 - 14:40 OS1031 The use of matrix-assisted laser desorption ionization - time-of-flight mass spectrometry (MALDI-TOF MS) for rapid bacterial identification in patients with smear- positive bacterial meningitis - a study of diagnostic accuracy Boaz Bishop*1, Yuval Geffen2, Ola Kassis3, Adi Kamar4, Mical Paul5, Ami Neuberger6 1Rambam Medical Center; Department of Internal Medicine B 2Rambam Health Care Campus; Microbiology Laboratory 3Technion 4Rambam Medical Center; Microbiology Laboratory 5Rambam Health Care Center; Division of Infectious Diseases 6Rambam Medical Center; Unit of Infectious Diseases Background: Bacterial meningitis is a rapidly occurring fatal disease, thus immediate diagnosis and adequate therapy are crucial. Diagnosis is obtained by cerebral spinal fluid (CSF); cultures results are the gold standard for diagnosis, but are available only within 24-72 hours. MALDI-TOF-MS identifies a wide range of bacteria by examining protein profiles. Little data exists regarding the use of MALDI- TOF-MS for examining CSF samples. We aimed at assessing the accuracy of MALDI-TOF-MS in rapid identification of CSF pathogens in patients with meningitis. Material/methods: We conducted a prospective study examining samples with positive CSF Gram stains with MALDI-TOF-MS. The results were compared with the CSF culture; identification was considered accurate when identical to the CSF culture results (species and genus level). Lab workers preforming the MALDI-TOF MS and ones interpreting its results were blinded to the result of the direct smear, besides the fact of it being positive. -
GRAM STAIN REAGENTS - for in Vitro Use Only - Catalogue No
GRAM STAIN REAGENTS - For in vitro use only - Catalogue No. SG51-55 Our Gram-Stain Reagents are intended to be The last step is the application of a counterstain. The used as a differential stain for the microscopic most common counterstain is safranin, which colors examination of bacterial cultures and laboratory decolorized cells pink. An alternate counterstain is basic specimens. fuchsin, which gives the decolorized cells more of a Gram staining is the single most useful test in bright pink or fuchsia coloration. The basic fuchsin the microbiology laboratory given its simplicity and counterstain works particularly well for anaerobic ability to differentiate bacteria into two main bacteria, but poorly for Legionella and Bordetella groups: gram-positive organisms and gram negative species. organisms. Hans Christian Joachim Gram first devised the original procedure in the late 19 th century, and although modifications have since been Formulation per Litre made, the basic principles and results remain the same. Our formulation is often referred to as the SG51 Gram Crystal Violet Hucker Modification. The staining spectrum includes almost all Crystal Violet ................................................ 20.0 g bacteria, many fungi, and parasites such as Ammonium Oxalate ....................................... 8.0 g Trichomonas , Strongyloides , and protozoan cysts. Methanol ................................................. 200.0 mL The notable exceptions include intracellular pathogens such as Chlamydia and Rickettsia , and SG52 Gram Iodine those organisms lacking a true cell wall such as Mycobacterium , Mycoplasma , and Ureaplasma . Iodine Crystals .............................................. 3.33 g The differential properties of the staining Potassium Iodide ........................................... 6.67 g process are attributed to the differences in composition between gram-positive and gram- SG53 Gram Decolorizer negative cell walls. -
DIFFERENTIAL STAINING, Part I
DIFFERENTIAL STAINING, Part I Differential staining is a procedure that takes advantage of differences in the physical and chemical properties of different groups of bacteria. It allows us to differentiate between different kinds of bacterial cells or different parts of a bacterial cell. I. GRAM STAIN The most commonly used differential stain is the Gram stain, first described in 1884 by Christian Gram, a Danish physician. The Gram reaction divides bacteria into two groups, those which are Gram-positive and those which are Gram-negative. Those organisms which retain the primary stain (crystal violet) are stained purple and are designated Gram-positive; those which lose the crystal violet and are subsequently stained by a safranin counterstain appear red and are designated Gram-negative. The conventional Gram-stain technique is described in the Procedure part of this handout; however, it is important to recognize early on that two aspects of the procedure are crucial: 1. The crystal violet treatment must precede iodine treatment. Iodine acts as a mordant, i.e., it increases the affinity of the cells for the crystal violet. Iodine alone has no bacterial staining capabilities. 2. Decolorization must be short and precise. Too long an exposure to 95% alcohol will decolorize Gram-positive as well as Gram-negative cells. The Gram stain has been used as a taxonomic tool for many years, aiding in the classification and identification of bacterial cells. However, it is also useful in a broader sense, as there appears to be a close correlation between the Gram reaction and many other morphological and physiological characteristics of bacterial cells. -
Differential Staining of Bacteria Microbiology Lab 3 Exercise
About Science Prof Online PowerPoint Resources • Science Prof Online (SPO) is a free science education website that provides fully-developed Virtual Science Classrooms, science-related PowerPoints, articles and images. The site is designed to be a helpful resource for students, educators, and anyone interested in learning about science. • The SPO Virtual Classrooms offer many educational resources, including practice test questions, review questions, lecture PowerPoints, video tutorials, sample assignments and course syllabi. New materials are continually being developed, so check back frequently, or follow us on Facebook (Science Prof Online) or Twitter (ScienceProfSPO) for updates. • Many SPO PowerPoints are available in a variety of formats, such as fully editable PowerPoint files, as well as uneditable versions in smaller file sizes, such as PowerPoint Shows and Portable Document Format (.pdf), for ease of printing. • Images used on this resource, and on the SPO website are, wherever possible, credited and linked to their source. Any words underlined and appearing in blue are links that can be clicked on for more information. PowerPoints must be viewed in slide show mode to use the hyperlinks directly. • Several helpful links to fun and interactive learning tools are included throughout the PPT and on the Smart Links slide, near the end of each presentation. You must be in slide show mode to utilize hyperlinks and animations. •This digital resource is licensed under Creative Commons Attribution-ShareAlike 3.0: http://creativecommons.org/licenses/by-sa/3.0/ Tami Port, MS Alicia Cepaitis, MS Creator of Science Prof Online Chief Creative Nerd Chief Executive Nerd Science Prof Online Science Prof Online Online Education Resources, LLC Online Education Resources, LLC [email protected] [email protected] From the Virtual Microbiology Classroom on ScienceProfOnline.com Image: Compound microscope objectives, T. -
Bacterial Cell Wall and Differential Staining
About Science Prof Online PowerPoint Resources • Science Prof Online (SPO) is a free science education website that provides fully-developed Virtual Science Classrooms, science-related PowerPoints, articles and images. The site is designed to be a helpful resource for students, educators, and anyone interested in learning about science. • The SPO Virtual Classrooms offer many educational resources, including practice test questions, review questions, lecture PowerPoints, video tutorials, sample assignments and course syllabi. New materials are continually being developed, so check back frequently, or follow us on Facebook (Science Prof Online) or Twitter (ScienceProfSPO) for updates. • Many SPO PowerPoints are available in a variety of formats, such as fully editable PowerPoint files, as well as uneditable versions in smaller file sizes, such as PowerPoint Shows and Portable Document Format (.pdf), for ease of printing. • Images used on this resource, and on the SPO website are, wherever possible, credited and linked to their source. Any words underlined and appearing in blue are links that can be clicked on for more information. PowerPoints must be viewed in slide show mode to use the hyperlinks directly. • Several helpful links to fun and interactive learning tools are included throughout the PPT and on the Smart Links slide, near the end of each presentation. You must be in slide show mode to utilize hyperlinks and animations. •This digital resource is licensed under Creative Commons Attribution-ShareAlike 3.0: http://creativecommons.org/licenses/by-sa/3.0/ Tami Port, MS Alicia Cepaitis, MS Creator of Science Prof Online Chief Creative Nerd Chief Executive Nerd Science Prof Online Science Prof Online Online Education Resources, LLC Online Education Resources, LLC [email protected] [email protected] From the Virtual Microbiology Classroom on ScienceProfOnline.com Image: Compound microscope objectives, T. -
Gram Stain Introduction
Gram Stain Introduction The Gram stain is a differential staining procedure used to categorize bacteria as Gram positive or Gram negative based on the chemical and physical properties of their cell’s walls. The bacteria are differentiated through a series of staining and decolorization steps. Gram positive cells will stain purple and Gram-negative cells will stain red to pink. Supplies and Reagents 1. Personal protective equipment (PPE) 3. 2. Slide rack 1. 3. Timer 4. Absorbent paper, such as bibulous paper 5. Water (tap water or deionized) 6. Crystal violet 7. Gram’s iodine 8. Decolorizer 9. Safranin (or carbol fuchsin) 5. 7. 10. Brightfield microscope with 100X objective 11. Immersion oil Instructions 1. Use appropriate PPE according to your laboratory’s procedures and safety manual. 2. Place the prepared fixed smear on a slide rack then flood the slide with crystal violet. 3. Wait at least 15 seconds* then rinse the slide with 9. 11. water. 4. Flood the slide with Gram’s iodine. 5. After 15 seconds* rinse the slide with water. 6. Apply the decolorizer to the slide. 7. Rinse the slide immediately with water. 8. Flood the slide with counterstain. 9. Wait at least 15 seconds* then rinse the slide with water. 10. Blot the slide with absorbent paper. Be careful not to wipe the cells off the slide. 11. Allow the newly stained slide to air dry completely. 12. View the slide under oil using the oil immersion objective for a total magnification of 1000X. 13. Record results based on your laboratory’s criteria. -
The Color of Tissue Diagnostics Routine Stains, Special Stains and Ancillary Reagents
The Color of Tissue Diagnostics Routine Stains, Special Stains and Ancillary Reagents The life science business of Merck KGaA, Darmstadt, Germany operates as MilliporeSigma in the U.S. and Canada. For over years, 100routine stains, special stains and ancillary reagents have been part of the MilliporeSigma product range. This tradition and experience has made MilliporeSigma one of the world’s leading suppliers of microscopy products. The products for microscopy, a comprehensive range for classical hematology, histology, cytology, and microbiology, are constantly being expanded and adapted to the needs of the user and to comply with all relevant global regulations. Many of MilliporeSigma’s microscopy products are classified as in vitro diagnostic (IVD) medical devices. Quality Means Trust As a result of MilliporeSigma’s focus on quality control, microscopy products are renowned for excellent reproducibility of results. MilliporeSigma products are manufactured in accordance with a quality management system using raw materials and solvents that meet the most stringent quality criteria. Prior to releasing the products for particular applications, relevant chemical and physical parameters are checked along with product functionality. The methods used for testing comply with international standards. For over Contents Ancillary Reagents Microbiology 3-4 Fixing Media 28-29 Staining Solutions and Kits years, 5-6 Embedding Media 30 Staining of Mycobacteria 100 6 Decalcifiers and Tissue Softeners 30 Control Slides 7 Mounting Media Cytology 8 Immersion