2,7-FLUORENEDIAMINE AND 2,5-FLUORENEDIAMINE AS PEROXIDASE REAGENTS FOR BLOOD SMEARS

  • 1 January 1976
    • journal article
    • research article
    • Vol. 88  (2) , 334-338
Abstract
New histochemical methods for peroxidase activity, which do not require benzidine as a hydrogen donor, were modified for hematological use. Diamine derivatives of fluorene (2,7-fluorenediamine and 2,5-fluorenediamine) were employed in place of benzidine and the result obtained was satisfactory. Two staining techniques were developed. Smears of peripheral blood or bone marrow aspirates were fixed in 2.5% glutaraldehyde solution for 1 min. Smears were then washed in tap water and covered either by a saturated solution of 2,7-fluorenediamine or by a 0.05% solution of 2,5-fluorenediamine in the presence of H2O2 for 5 min. After being washed in running tap water, they were counterstained with Carazzi''s hematoxylin solution for 10 min. The 2nd technique employs pretreatment of blood smears with 5% CuSO4 solution and aldehyde fixation was omitted. Smears were counterstained by Giemsa stain. The nuclear chromatin structures were well preserved and nucleoli were easily distinguished in the immature cells. In the 1st method, clear brown granules were recognized in the cytoplasms. In the 2nd method, peroxidase-positive granules were stained black and the staining of nucleus and cytoplasm resembled that of McJunkin''s method.

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