Home – Home – Jornals – Siberian Scientific Medical Journal 2019 number 6

Aim of the study was to compare the distinct types of progressive hematoxylin stains and to optimize the protocols of hematoxylin and eosin staining of blood vessels, heart, liver and spleen. Material and methods. Heart (ventricles), abdominal aorta, liver (right lobe) and spleen (left part) of the Wistar rats were excised, fixed in 10% neutral phosphate buffered formalin for 24 h, washed in tap water for 2 h, dehydrated in ascending ethanol series (70 %, 80 %, and 95 %) and isopropanol, embedded into paraffin and then sectioned (5 μm) using rotary microtome. Staining was performed using Mayer’s, Gill’s, or Carazzi’s hematoxylin during 2, 5, or 15 minutes and 1 % alcoholic/aqueous eosin for 2 minutes without differentiative solution. Results were assessed by three independent histologists. Results. All examined progressive hematoxylin stains had their distinctive features. Mayer’s hematoxylin demonstrated the most intensive nuclear staining; however, staining for 15 minutes could lead to the bluing of cytoplasm and extracellular matrix. In contrast, Gill’s hematoxylin was characterized by less intensive nuclear staining and achieved clear blue-violet shade only after 15 minutes of staining. Carazzi’s hematoxylin showed balanced coloration of nuclei and cytoplasm/extracellular matrix and did not change the red/pink shades of eosin, yet the intensity of nuclear staining was less as compared to Mayer’s hematoxylin. Short-term (2 minutes) staining was insufficient to reach intensive nuclear staining. Conclusion. The optimal hematoxylin and eosin staining protocol is to use eosin for 2 minutes following staining by Carazzi’s hematoxylin for 15 minutes (for aorta), Carazzi’s or Gill’s hematoxylin for 15 minutes or Mayer’s hematoxylin for 5 minutes (for liver), Carazzi’s or Gill’s hematoxylin for 15 minutes (for heart), and Carazzi’s hematoxylin for 5 minutes (for spleen).