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Question
staining - capsule staining
steps
use inoculating loop to aseptically mix bacterial sample with primary stain.
apply primary stain (india ink or nigrosin) to a clean slide.
create a thin smear by dragging a clean slide through the sample.
allow sample to thoroughly air dry.
rinse with water and prepare for microscopy.
apply secondary stain (crystal violet) to smear.
Identify the staining method and initial step
The header indicates "STAINING - CAPSULE STAINING". Capsule staining is a negative staining technique where the background is stained first. Therefore, the first step is to apply the primary acidic stain (India ink or nigrosin) directly to a clean slide.
Determine the sequence of smear preparation
Once the primary stain is on the slide, the bacterial sample is added and mixed aseptically using an inoculating loop. Next, a second clean slide is used to drag the mixture across the slide to create a thin, even smear.
Determine the final staining and preparation steps
The smear must then be allowed to thoroughly air dry (heat-fixing is avoided as it shrinks the capsules). After air drying, the secondary basic stain (crystal violet) is applied to color the bacterial cells inside the capsules. Finally, the slide is gently rinsed with water and prepared for microscopy.
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| No. | Answer |
|---|---|
| 2 | Use inoculating loop to aseptically mix bacterial sample with primary stain. |
| 3 | Create a thin smear by dragging a clean slide through the sample. |
| 4 | Allow sample to thoroughly air dry. |
| 5 | Apply secondary stain (crystal violet) to smear. |
| 6 | Rinse with water and prepare for microscopy. |