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Lab Methods: Brightfield Microscopy For the Diagnosis of STIs
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Lab Methods: Brightfield Microscopy For the Diagnosis of STIs

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The PowerPoint presentation, "Brightfield Microscopy For The Diagnosis Of STDs" is from the STD/STI Laboratory Methods Series. It provides basic laboratory knowledge and skills associated with ...

The PowerPoint presentation, "Brightfield Microscopy For The Diagnosis Of STDs" is from the STD/STI Laboratory Methods Series. It provides basic laboratory knowledge and skills associated with microscopic examinations for the diagnosis of sexually transmitted diseases and/or infections (STD/STI). The target audience includes physicians, nurse practitioners, nurse midwives, nurse clinicians, physician assistants, laboratorians, medical students, and any medical/health personnel new to Brightfield microscopy (or in need of refresher Brightfield microscopy course).

The presentation, "Brightfield Microscopy For The Diagnosis Of STDs" was designed to inform and instruct clinical and non-clinical medical personnel on the proper laboratory procedures associated with saline and potassium hydroxide (KOH) wet preparations used in the identification of Clue Cells (Bacterial vaginosis), Trichomonas vaginalis, and Candida species. It was prepared by Sally S. Cherry, MT(ASCP) for the exclusive use in STD/STI Laboratory Methods Training Courses and Workshops conducted through Cherry Consulting Network. After the “Brightfield Microscopy For The Diagnosis Of STDs” training course, participants will be able to… Identify the major components of the compound microscope; Demonstrate proper techniques in the care and use of the compound microscope; Describe the recommended laboratory procedures for the collection of genital specimens; Demonstrate proper microscopic procedures for specimen preparation; Identify trichomonads in saline solution. Identify clue cells in saline solution; and Identify Candida species in KOH solution.

Please note that written permission from the author must be obtained before using, reproducing, or photocopying this STD/STI Laboratory Methods Series PowerPoint presentation. To make arrangements or to register for a STD/STI Lab Methods Training/Refresher Course, contact Sally S. Cherry, MT(ASCP) at (443) 212-8262 or email: scherry98@gmail.com

Presenter Bio
As a medical technologist certified by the American Society of Clinical Pathologists (ASCP), Sally S. Cherry, MT(ASCP) serves as a national and international laboratory consultant specializing in the laboratory diagnosis of sexually transmitted diseases (STDs) and sexually transmitted infections (STIs). In this role, Ms. Cherry provides basic laboratory training, in-service training, and train-the-trainer for clinical and non-clinical medical personnel. The topics of instruction include Brightfield Microscopy: Use and Care of the Compound Microscope; Brightfield Microscopy for the Identification of Sexually Transmitted Diseases and/or Infections (STD/STI); and Darkfield Microscopy for the Identification of Treponema pallidum, the causative agent of syphilis. In addition, Ms. Cherry provides laboratory consultations related to quality control, lab management, performance reviews, and other related tasks.

Website: http://cherrynetwork.weebly.com/ms-bugladys-world.html

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Lab Methods: Brightfield Microscopy For the Diagnosis of STIs Document Transcript

  • 1. 1 BRIGHTFIELD MICROSCOPY FOR THE DIAGNOSIS OF STDs By Sally S. Cherry, MT(ASCP) Laboratory Consultant & Instructor Cherry Consulting Network BRIGHTFIELD MICROSCOPY FOR THE DIAGNOSIS OF STDs MICROSCOPIC DIAGNOSIS OF TRICHOMONIASIS, CANDIDIASIS & BACTERIAL VAGINOSIS Please note that written permission from the author must be obtained before using, or copying this PowerPoint laboratory presentation. Objective: The STD laboratory presentation is designed to provide the knowledge and skills to properly perform saline and potassium hydroxide (KOH) wet preparations used in the identification of: Clue Cells (Bacterial vaginosis) Trichomonas vaginalis Candida species
  • 2. 2 Recommended Equipment Brightfield Microscope • Eyepieces (Oculars) • Body Tube • Arm • Objectives (10X-40x-100x) • Condenser, Brightfield • Iris Diaphragm Lever • Stage • Fine Focus Adjustment Knob • Coarse Focus Adjustment Knob • Light (Spare Light Bulb) • Base Recommended Supplies Examination Room Specimen Collection • Gloves • Swabs, Cotton, or Dacron • pH Paper • Gauze (Squares) • Microscope Slides • Cover Slips, Glass • Saline Solution • Potassium Hydroxide (10%) Saline Wet Preparations Supplies Needed • Latex Gloves • Sterile Pipette or dropper • Small Test Tube - 12 x 75 mm • Normal Saline (0.5 - 1.0 ml) • Tube label, or ID marker
  • 3. 3 Saline Wet Preparations Test Tube Method Prepare Saline Tube • Add .5 - 1.0 ml saline to tube. • Cover with cotton plug, or cap. Saline Wet Preparations Specimen Collection & Handling Prepare Saline Emulsion • Collect vaginal specimen with swab. • Check and note pH of specimen. • Place swab immediately into saline. • Tap tube to wash off specimen (“milky”). • Label tube with patient’s ID • Leave swab in saline tube. Wet Preparations Brightfield Microscopic Exam • Brightfield Microscope 10X Objective 40X Objective Condenser (lowered) • Microscope Slide • Cover Slip • Saline Tube: Specimen
  • 4. 4 Saline Wet Preparations Examine Specimen for:  Trichomonas vaginalis  Clue Cells (Bacterial vaginosis)  Yeast Cells and Hyphae Saline Wet Preparations Trichomonas vaginalis • Pear-shaped protozoan • Motile; jerky movement • Directional movement • Undulating membrane • Four (4) anterior flagella • One (1) posterior flagella • One (1) nucleus (Illustration of Trichomonas vaginalis) Saline Wet Preparations Trichomonas vaginalis (Note: Motility can not shown here.)
  • 5. 5 Saline Wet Preparations Identifying Clue Cells for BV... Clue, or Epithelial cell? How are the cells different? Clue, or Epithelial cell? Clue Cell vs Epithelial cell How are the cells different? Epithelial cell… Fine “clean” appearance No bacteria present on surface Smooth edges Clue cell… Granular appearance Bacteria present on surface Irregular edges Saline Wet Preparation Clue Cells… • Epithelial cells • Granular appearance • Presence of adherent bacteria • Irregular edges
  • 6. 6 Saline Wet Preparation Identifying the Clue Cell… 1. Is it an Epithelial cell? Yes or No 2. Look for…  Irregular edges  Granular appearance on surface  Presence of bacteria You call it… Clue Cell? Epithelial Cell? Clue Cell? Epithelial Cell?
  • 7. 7 Clue Cell (Gram-Stained Specimen) Gram-negative adherent bacteria present. (Gram-positive bacteria may also be present.) Troubleshooting: Problem #1 1. What do you observe in this specimen? 2. How would you eliminate this problem? Troubleshooting: Solution #1 Observation: Oil Droplets Recommended Action: • Check cover slip for oil • Check objective for oil • Clean 40X objective • Prepare and examine a second specimen
  • 8. 8 Troubleshooting: Problem #2 1. What do you observe in this specimen? 2. How would you eliminate this problem? Troubleshooting: Solution #2 Observation: Air Bubble Recommended Action: • Check cover slip for air bubble • Examine “specimen” area • Prepare a second specimen using more “specimen” (saline emulsion). • Examine a second specimen KOH Wet Preparation (10% Potassium Hydroxide) Prepare KOH Slide • Tap tube to mix specimen. • Add specimen to slide. • Add a drop of 10% KOH. • Apply cover glass. • Examine for yeast & hyphae.
  • 9. 9 KOH Wet Preparation Candida albicans • Oval Budding Yeast Cells • Branching Hyphae KOH Wet Preparation Budding Yeast Cells Branching Hyphae
  • 10. 10 KOH Wet Preparation Yeast: Non-Branching Hyphae KOH Wet Preparation Yeast Cells & Hyphae KOH Wet Preparation Yeast Cells & Hyphae • Budding Yeast Cells • Branching Hyphae
  • 11. 11 Is this worth reporting? Yeast Cells & Hyphae (Gram Stain) Yeast Cells stain Gram- positive. Cherry Consulting Network P.O. Box 211 Joppa, MD 21085 (Email: scherry98@gmail.com) Please note that written permission from the author must be obtained before using, reproducing, or photocopying this PowerPoint laboratory presentation. (Presentation Revised: November 2006) “Have Microscope, Will Train”