The surrounding membrane of the granulations of the neutrophil and its acid alcohol fastness.

By PAULO SEABRA W lIEN S 1’AIXIXG A BLOOI) SMEAR for oxidase by metinoc! of Loele and Seabra (see Appendix: Tecinnic’s) tine smear is fixed by formahin vapor tinen treated with a-naphntinol and finally with gent-ian violet. At tinose sites where the enzyme is present, tine a-mnaplnthol is oxidized. It then acts as a mordantwhicin fixes the gentian so that those parts of the cells containing oxidase are stained blue. One can c)bserve that only tIne cosinophils and neufropinils am’e rich in oxidase. Occasionally some neutrophils effuse a certain quantity of this enzyme w hich may he noted in contiguous eryt-inrocytes (fig. 1 a). An investigation was undertaken to exl)lain nviny some of the netttrophils passed on tineir oxiclase to the nearby eryt-hrocytes in such an intense way and otiners did not. If tine cf’fusion of tine neutrophilic oxida-se were a mere dissolution of tine enzyme clue to tine lidluids used in fine staining process all of the granulations should h)e dissolved. Consec uenfly, itwas postulated! tinat f-he gn’anulations consisted of an enzymatic liquid, protected! by a surroumnding membrane whicin is sul)ject to a process of maturation,t This membrane upon reaching maturation (hoes not resist tine contract-ion produced by the formol vapors used to fix f-he smear, and t-Inet’efom’e ruptures during tinis pinase of f-he process. \Vinen c’ovel.imng the slide with the liquid! “mordant,” tine enzyme of f-he nipt-um’ed granulation is diluted. I)uring tinis 1)t’OcesS, it may also be transported to adijacentsites and! eventually be adlsol’hed! to fine surface of contiguous erythrocvtes. Occasionally itis possii)le to obsem’ve unstained areas in the neutm’opinils presumably fom’mecl by tine empty granulations (fig. 1 b) . From the meniscus thus formed, the conclusid)n follows that the oxidase liquid must i)e very thick and gelatinous (fig. 2). Rei)uc’k,2 tinm’ough tine use of the electron microscope, has greatly strengthened tine concepttinat neutropinilic granulations are not hemogenous solid bodies like concretiomns. In his photographs of fine neutrophile granulations thney appear as liquid spineres surrounded by a membm’ane since tiney are more transparent in the center tlnamn at tine pel.ipinery. Thei’efoi’e, it is to i)e expected tinat if the 1)100(1 were decompressed m’apidly and violently, not only would f-inc meml)ranes thatmad reached matun’afion be bm’oken buttine immature ones would also i)e ruptured. This hsas been observed durinng the pi’oeess of decompressing laboratory animals, or dum’ing tine drawing of blood during flights at inigh altitudes.3 The mcmi)rane of the l)roken and empty granulation free fm’om any oxiclase content may be d!emonnstm’ated by simply cmusining the neutrophils. TinIs can he clone when makinng the smear by pressing one slide against tine othem’ (fig. 3). In A are seen granulations scattered near the neutrophils but retaining their oxidase con-