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Easy Guide to Simple Stain Preparation for Microbial Smear Procedures

This guide details the step-by-step process for preparing a simple stain for microbial examination. It covers three main procedures: preparing a smear from broth culture using TSB, creating a smear from solid media using TSA, and preparing a smear of an unknown sample. Key techniques include resuspension, aseptic transfer, and heat fixing. The staining options include Methylene Blue and Crystal Violet, followed by rinsing and drying. These procedures are essential for observing microbial morphology and arrangement.

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Easy Guide to Simple Stain Preparation for Microbial Smear Procedures

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  1. Ex. 7 Preparing a Simple Stain

  2. Procedure # 1 • Prepare a Smear from Broth Culture - TSB • Resuspend Broth Culture • Aseptic Transfer Technique • Heavy Turbidity – 1 loopful • Light Turbidity – 2 – 6 loopfuls • Air Dry • Heat Fix

  3. Procedure # 2 • Prepare a Smear from a solid media - TSA • Add sterile H2O • Add Bacteria • Air Dry • Heat fix

  4. Procedure # 3 • Prepare a smear of an Unknown - TSB • Air Dry • Heat fix

  5. Simple Stain • Methylene Blue • Crystal Violet • 1. Add Stain (1 minute) • 2. Rinse with water • 3. Blot dry with Bibulous paper • Observe Morphology and arrangement

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