H&E staining of pond water sample

This is my first attempt to do H&E staining – I used the pond water sample from John Brockley’s garden yesterday which showed so much Volvulus. Heat fixed it first then applied the Haematoxylin for around 20 mins, washed it off, and then eosin for 5 mins. The former should bring out nuclear material in purple and the latter cytoplasmic material – I will leave you to decide whether I was successful from the photos below!

Haematoxylin and eosin stain is one of the principal stains in histology. It is the most widely used stain in medical diagnosis and is often the gold standard; for example when a pathologist looks at a biopsy of a suspected cancer, the histological section is likely to be stained with H&E and termed “H&E section”, “H+E section”, or “HE section”. A combination of hematoxylin and eosin, it produces blues, violets, and reds (Wikipedia).

I used this staining technique a lot in my anatomy degree and most pathology slides use H&E.


For comparison non-stained photos from yesterday from same water sample (below – these first two are NOT stained with H&E):

Volvulus-from-John-Brockleys-pond-270817-HE-stain-10x-Planapo-obj-Zeiss-Photomic-III (below – reds and purples are evident but not as pronounced as I expected. However this is only using the 10x objective so higher magnification would be more insightful into whether stain is effective):


16x objective H&E stain:

40x objective H&E – panorama of 7 photos (below – try downloading this image and zooming in – it is a much bigger image than previous ones as it is created from 7 photos – there is a great amount of detail within the cytoplasms of the cells other than chloroplasts (which were visible in unstained images)):

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