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Here's when each modifier applies: No modifier needed: the injection is the only service provided at the visit Modifier 25 needed: a separately identifiable E/M service is performed the same day Modifier 59 needed: multiple distinct injections are given at the same visit involving different drugs at different sites Modifier 76 needed: the same injection is repeated by the same provider on the same date of service Modifier 77 needed: a different provider repeats the same injection on the same date One warning that trips up a lot of billers: appending Modifier 59 to an E/M service code is incorrect

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Tissue collection and microscopic analysis Spleens were fixed in 4% formaldehyde (methanol-free paraformaldehyde, Thermo Scientific #28908) overnight at 4 °C, washed and transferred into phosphate-buffered saline containing 0.02% sodium azide and 20% sucrose and then cut with a microtome (CryoStar NX 50, Thermo Scientific) to obtain 8 µm tissue sections 94 that were then stained for 10 min with 0.3 µM 4′,6-diamidino-2-phenylindole in Dulbecco’s phosphate-buffered saline (Thermo Scientific #62247)