To stain tissue sections using Cresyl Violet for histological analysis (e.g., infarct volume quantification), involving a series of dehydration, staining, and clearing steps.
Classification
Sample typeTissue
TechniqueMicroscopy & Imaging
Experimental contextIn vitro
Biological modelHumanMouseRat
Otherhistological staining
Reproducibility status
Reproducible
No reproducibility blockers or risks reported. The protocol runs end-to-end.
12 healthy
⏱ Total 37 min1 section12 steps
Cresyl Violet Staining Procedure
37 min
1
Decongelate the tissue sections on microscope slides.
2
Immerse slides in 100% alcohol for 2 minutes.2 min
3
Immerse slides in 85% alcohol for 2 minutes.2 min
4
Immerse slides in 70% alcohol for 2 minutes.2 min
5
Rinse slides in distilled water (H2Od) for 2 minutes.2 min
6
Stain slides with Cresyl Violet solution (1 mg/ml of CV (Sigma) in distilled water and 0.3 % of glacial acetic acid (Carlo Erba)) for 10 minutes.15 min
Critical stepStaining time must be carefully controlled; under-staining leads to faint nuclei, over-staining causes excessive background.
7
Rinse slides in distilled water (H2Od) for 1 minute, repeat twice.1 min
8
Dehydrate slides in 70% alcohol for 1 minute.1 min
9
Dehydrate slides in 85% alcohol for 1 minute.1 min
10
Dehydrate slides in 100% alcohol for 1 minute.1 min
11
Clear slides in Xylene for 7 minutes.10 min
Critical stepEnsure complete clearing; insufficient clearing results in cloudy or opaque sections.
12
Mount coverslips using mounting medium (e.g., Eukitt).
Critical stepApply mounting medium evenly and avoid bubbles to preserve sample integrity and optical clarity.
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