LSB466 Week 8 Pimp My Slide PDF

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LSB466: Week 8 Pimp my slide How to improve your staining technique

1 LSB466: Week 8, Semester 2, 2018.

Medical Sciences, QUT. CRICOS No.00213J

Prac Assessment (50%) Student portfolio (20%) 5 “Best” slides: due end of Week 10 Thursday prac class Unknown slides: due end of Week 12 prac class

Staining Exam (30%) Held in Week 13 prac class

2 LSB466: Week 8, Semester 2, 2018.

Medical Sciences, QUT. CRICOS No.00213J

5 best slides • To be submitted by end of Week 10 Thursday prac class (along with brief report). – Report should briefly explain why you have chosen each slide (200-400 words per slide).

• A portfolio of 5 slides displaying best examples of your staining techniques.

3 LSB466: Week 8, Semester 2, 2018.

Medical Sciences, QUT. CRICOS No.00213J

5 best slides • • • • •

A H&E stained slide. A PAS stained slide. A Van Gieson stained slide. An elastin stain (VVG or Orcein) A slide stained with either G&S, MT or MSB.

4 LSB466: Week 8, Semester 2, 2018.

Medical Sciences, QUT. CRICOS No.00213J

Unknown slides • 5 sets of two slides handed out in week 9. • Perform H&E stain to identify tissues. • Perform special stain based upon tissue ID. • To be submitted by end of week 12 prac class. 5 LSB466: Week 8, Semester 2, 2018.

Medical Sciences, QUT. CRICOS No.00213J

Staining Exam • To be held during week 13 prac class. • ~8 slides to be stained in 2.5 hours (~5 different stains). • Details to be announced in Week 12. • Will have one week to prepare your strategy. • Slides to be submitted by end of Week 13 prac class. 6 LSB466: Week 8, Semester 2, 2018.

Medical Sciences, QUT. CRICOS No.00213J

Example Staining Exam

7 LSB466: Week 8, Semester 2, 2018.

Medical Sciences, QUT. CRICOS No.00213J

Grade

Slide marking criteria

7

Demonstrates well purpose of staining method including counterstain (if required). No significant presentations issues.

6

The slide has been well stained but with major presentation issues that while essentially cosmetic in nature detract from the overall quality. Main purpose of stain achieved. BUT Quality of stain could be improved (e.g. inadequate differentiation) or counterstaining poor.

5

4

Main purpose of stain achieved, but with room for improvement. AND/OR Failure to demonstrate counterstaining. Major presentation issues potentially interfering with diagnosis or data collection.

3

Presently unable to demonstrate ability to conduct stain in a manner that can be used to support an accurate diagnosis/result.

How to improve slides • Repeat stains as often as possible. • make use of revision weeks.

• Physical damage to sections impossible to correct. • Many staining errors can be corrected if have time. 9 LSB466: Week 8, Semester 2, 2018.

Medical Sciences, QUT. CRICOS No.00213J

The“H&E” stain • Use microscope control to ensure Hx staining correct.

•

Promptly rinse slides in alcohols during dehydration. 10

LSB466: Week 8, Semester 2, 2018.

Medical Sciences, QUT. CRICOS No.00213J

PAS • Generally

performed well. • Potential issues: – reagents applied to

wrong side of slide. – Mayer’s Hx too dark (10 seconds max) and “blue”. 11 LSB466: Staining Principles, 2018, D. Harkin

Medical Sciences, QUT. CRICOS No.00213J

Van Gieson Stain • Hx

• 1 dip in acid alcohol

• Acid fuchsin • •

2 min max avoid water

• Picric acid •

blot and briefly rinse in 100% ethanol.

• Take care with blotting! LSB466: Week 5, Semester 2, 2018, D. Harkin

Medical Sciences, QUT. CRICOS No.00213J

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Blotting artefact • Caused by accidental side-ways movement while blotting. • Presents as characteristic “pinched” areas of tissue. • No way to correct.

13 LSB466: Week 5, Semester 2, 2018, D. Harkin

Medical Sciences, QUT. CRICOS No.00213J

Masson’s Trichrome Stain • Post-fix slides for best results. • Hx: 1 dip acid alcohol. • Masson’s red: 2 min • PMA – requires effort. • Methyl blue: 2 min • don’t over rinse with acetic acid. • can blot if necessary. 14 LSB466: Week 5, Semester 2, 2018, D. Harkin

Medical Sciences, QUT. CRICOS No.00213J

Masson’s Trichrome Stain

15 LSB466: Week 5, Semester 2, 2018, D. Harkin

Medical Sciences, QUT. CRICOS No.00213J

Can Blot Methyl Blue • If having trouble retaining methyl blue.

• Brief rinse acetic acid (~1 mL) • Then blot.

• Use same approach as for blotting Van Gieson. 16 LSB466: Week 5, Semester 2, 2018, D. Harkin

Medical Sciences, QUT. CRICOS No.00213J

MSB Stain • Issues generally similar for Masson’s trichrome. • Post-fix • Don’t overdifferentiate Hx. • PTA easier to use than PMA. • Methyl blue: 2 min • Blot to retain better methyl blue. LSB466: Week 5, Semester 2, 2018, D. Harkin

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Medical Sciences, QUT. CRICOS No.00213J

Verhoeff’s Elastin Stain • Will compare with Orcein stain but VVG more likely to be used in staining exam due to time.

18 LSB466: Week 5, Semester 2, 2018, D. Harkin

Medical Sciences, QUT. CRICOS No.00213J

Verhoeff’s Elastin Stain • Issues: – stain upside down in Petri dish.

– differentiate well in ferric chloride. – VG counterstain: • See VG issues.

19 LSB466: Week 5, Semester 2, 2018, D. Harkin

Medical Sciences, QUT. CRICOS No.00213J

Learning Objectives • Make time to review your stained slides. • Attend multi-header sessions when •

scheduled to obtain feedback. Practice tissue ID using class sets of stained slides and via Histopics.tumblr.com

• Memorise staining protocols as much as possible:

•

remember can bring notes into staining exam but not practical to use staining manual.

LSB466: Week 8, Semester 2, 2018, D. Harkin

Medical Sciences, QUT. CRICOS No.00213J

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